Podcasts about hla dr

In this episode, I address ALL of the questions I get about mold, including:Can you get better living in it?Do you need all of the supplements?How to detox without symptoms?HLA/DR and geneticsAnd so much more!Also, I share my experience personally of at least 3 exposures and how they are all different, what I see clinically, and what mold sheds a light on when it comes to our foundational health!I also speak a lot about water (not just the water we drink), but how the water and the mitochondria influence our genetic expression. So to truly support our genetics, we have to support what controls those genes being turned on or off.Here is the water episode I mentioned.For personalized testing to look at toxin load, etc, click HERE.Resources mentioned in this episode:Mycircadian APP DOCTOR (code)Ra Optics (Code DRSTACYND)Bon Charge (Code DRSTACY) red light panel and circadian bulbsHigher Dose (my FAV sauna blanket with low to no EMF) code DRSTACYCGM (Code DRSTACYND)Analemma Water (structuring)Spring Aqua (my FAV water system) Thank you all so much for being here! For more from me, visit my IG @dr.stacy.nd or my site www.dr-stacy.com-Dr. StacyThis information is just that; information only - not to be taken as medical advice. Please contact your primary care before changing anything to your routine. This information is not mean to diagnose, treat, or cure disease.

Themes: CIRS, Mold Illness, Brain Retraining Show: Have you dealt with water damage in your house or symptoms related to mold illness? Today's guest, Katie Nickerson, shares her mold journey and healing tools using the structure of brain retraining. Katie is a brain retraining coach, helping clients access their innate healing wisdom to rewire their brains and thrive. Integrating neuroscience, nervous system regulation, somatic tools, and parts work. After years of mystery illness and multiple diagnoses, including CIRS, CFS/ME, MCAS, chronic pain, and more, Katie's health declined despite rigorous healing protocols. Listen to this episode to hear how her turnaround came with brain retraining, which gave her the tools to rewire her brain and make a full recovery from chronic illness!  Show Notes:  - Feeling stuck with CIRS symptoms and discovering brain retraining as a way to heal  - Brain retraining isn't just a modality; it's a process to improve the immune system to set up your body to heal - Katie's symptoms going unresolved after years of trying the Shoemaker protocol - The HLA-DR gene and how it affected Katie's approach to treatment - Talking to a stranger at the doctor's office led Katie to brain retraining - Katie's trip to Florida and how it changed her belief system on how to heal - Healing is not linear   Resources: -Katie's Instagram: @abodehealingco -Katie's Free Guide - Retraining Your Brain Around Mold -Schedule a Free Discovery Call for 1-1 Coaching -Healing From Mold Sensitivity Course -Lindsay's Instagram: @myvitalside or TikTok: @myvitalside -Visit Lindsay's Website -Click here for Lindsay's FREE Brain Retraining Guide  -Cait's Instagram: @caitmurphy.co and visit Cait's Website -Click here for Cait's FREE Masterclass

In this episode, we dive into the critical role of HLA gene variants, such as HLA-DRB1, and their profound impact on immune system function, particularly focusing on their significance in conditions like Lyme disease and mold toxicity. We'll explore how variations in these HLA genes, like the HLA-DRB1*0401 allele, influence the body's ability to recognize and respond to pathogens, potentially leading to chronic symptoms. The discussion will also cover HLA gene variant testing and how it can lend insight when it comes to taking a bioindividual approach to supporting the immune system. Topics: 1. Introduction - Explanation of HLA gene variants - Importance of HLA genes in immune system function 2. Basics of Genetics and Immunology - What is a chromosome? - Introduction to genes and their functions - Overview of DNA and protein synthesis 3. Major Histocompatibility Complex (MHC) - Description of the Major Histocompatibility Complex - Division of MHC genes into Class I, II, and III - Role of Class I MHC genes in antigen presentation - Role of Class II MHC genes in antigen presentation - Function of Class III MHC genes 4. HLA Genes and Immune Function - Explanation of Human Leukocyte Antigen (HLA) genes - Connection between MHC genes and HLA genes (human MHC) - Transcription and regulation of HLA genes - Response to internal and external signals - Role of Antigen-presenting cells (APCs) 5. Pathogen Recognition and Immune Activation - Mechanism of pathogen recognition by HLA genes - Activation of immune response by antigen presentation - Formation and function of peptide-HLA complexes 6. Impact of HLA Variants on Disease - Specific focus on Lyme disease and mold toxicity - Role of HLA Class II alleles in immune response - Association of HLA-DR alleles with chronic Lyme disease - Sensitivity to mold exposure linked to certain HLA gene variants 7. Testing and Implications of HLA Gene Variants - Importance of HLA gene variant testing - Broader implications for diseases like lupus and multiple sclerosis - Personalized approaches based on HLA gene variants 8. Biotoxin Illness and Poor Antigen Presentation Due to an HLA Gene Variant - Strategies to support immune system function in the presence of biotoxins - Importance of detoxification pathways and binders Thank you to our episode sponsor: ⁠⁠⁠⁠⁠Liver Medic⁠⁠⁠⁠⁠⁠ Use code Chloe20 to save 20% on ⁠⁠⁠⁠⁠⁠⁠"Leaky Gut Repair"⁠⁠⁠⁠⁠⁠⁠ ⁠⁠⁠⁠⁠⁠⁠Brendan's YouTube Channel⁠⁠⁠⁠⁠⁠⁠ ⁠⁠⁠⁠⁠⁠⁠https://x.com/livermedic⁠⁠⁠⁠⁠⁠⁠ Thanks for tuning in! Get Chloe's Book Today! "⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠75 Gut-Healing Strategies & Biohacks⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠" Follow Chloe on Instagram ⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠@synthesisofwellness⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠ Follow Chloe on TikTok @chloe_c_porter Visit ⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠synthesisofwellness.com⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠ to purchase products, subscribe to our mailing list, and more! --- Support this podcast: https://podcasters.spotify.com/pod/show/chloe-porter6/support

BUFFALO, NY- April 16, 2024 – A new #researchpaper was #published in Oncotarget's Volume 15 on April 12, 2024, entitled, “Novel therapeutic bispecific antibodies for B-cell lymphoma targeting IgM and other antigens on the B-cell surface.” The B-cell receptor regulates B-cell proliferation and apoptosis. Aberrations in BCR signaling are associated with the development and progression of B-cell malignancies, such as mantle cell lymphoma, diffuse large B-cell lymphoma, and chronic lymphocytic leukemia, many of which express the IgM type of BCR on their cellular surface. Therefore, IgM is an attractive target for therapeutic antibodies against B-cell malignancies. However, soluble IgM competitively binds to anti-IgM antibodies in the serum, and these antibodies show insufficient cytotoxic activity. Thus, antibody therapy targeting IgM is hindered by the presence of soluble IgM in the blood. In this new study, researchers Takahiro Ohashi, Sayuri Terada, Shinsuke Hiramoto, Yuko Nagata, Hirokazu Suzuki, Hitoshi Miyashita, Tetsuo Sasaki, Yasukatsu Tsukada, and Keiko Fukushima from ZENOAQ (Zenyaku Kogyo Co., Ltd.) used a bispecific antibody to address this problem. “In this study, we aimed to produce IgM-dependent bispecific antibodies targeting IgM and the other B-cell antigens such as CD20, CD32b (FcγRIIB), CD79b, and human leukocyte antigen (HLA)-DR using the Cys1m technology [10, 43–45]. Additionally, the correct IgG-like bispecific antibody structures were confirmed and their efficacies in the presence of soluble IgM were analyzed.” The researchers generated bispecific antibodies bound to IgM and other B-cell antigens such as CD20 and HLA-DR using their own bispecific antibody-producing technology, Cys1m. These bispecific antibodies directly inhibited cell proliferation via cell-cycle arrest and apoptosis in vitro, although large amounts of soluble IgM were present. Additionally, a bispecific antibody bound to IgM and HLA-DR (BTA106) depleted B-cells in cynomolgus monkeys. “These data suggest that anti-IgM/B-cell surface antigen-binding specific antibodies are promising therapeutic agents for B-cell malignancies. Moreover, the bispecific antibody modality can potentially overcome problems caused by soluble antigens.” DOI - https://doi.org/10.18632/oncotarget.28578 Correspondence to - Keiko Fukushima - keiko_fukushima@mail.zenyaku.co.jp Sign up for free Altmetric alerts about this article - https://oncotarget.altmetric.com/details/email_updates?id=10.18632%2Foncotarget.28578 Subscribe for free publication alerts from Oncotarget - https://www.oncotarget.com/subscribe/ Keywords - cancer, bispecific antibody, Cys1m, IgM, lymphoma, cynomolgus monkey About Oncotarget Oncotarget (a primarily oncology-focused, peer-reviewed, open access journal) aims to maximize research impact through insightful peer-review; eliminate borders between specialties by linking different fields of oncology, cancer research and biomedical sciences; and foster application of basic and clinical science. Oncotarget is indexed and archived by PubMed/Medline, PubMed Central, Scopus, EMBASE, META (Chan Zuckerberg Initiative) (2018-2022), and Dimensions (Digital Science). To learn more about Oncotarget, please visit https://www.oncotarget.com and connect with us: Facebook - https://www.facebook.com/Oncotarget/ X - https://twitter.com/oncotarget Instagram - https://www.instagram.com/oncotargetjrnl/ YouTube - https://www.youtube.com/@OncotargetJournal LinkedIn - https://www.linkedin.com/company/oncotarget Pinterest - https://www.pinterest.com/oncotarget/ Reddit - https://www.reddit.com/user/Oncotarget/ Spotify - https://open.spotify.com/show/0gRwT6BqYWJzxzmjPJwtVh Media Contact MEDIA@IMPACTJOURNALS.COM 18009220957

Why You Should Listen:  In this episode, you will learn about the root causes of CIRS including Actinos and endotoxins. About My Guests: My guest for this episode is Dr. Eric Dorninger.  Eric Dorninger ND, LAc is a Registered Naturopathic Doctor and Licensed Acupuncturist. He graduated from Bastyr University, the leading accredited university for science-based natural medicine.  Prior to medical school, he received his B.A. in Kinesiology from the University of Colorado, Boulder in 1997. During this time he also finished his E.M.T. (Emergency Medical Technician) training and volunteered at Porter Care Hospice in Denver. This dual exposure of medical perspectives laid down the roots for Dr. Dorninger's integrated approach to diagnosis, treatment, and healing.  Following undergrad, Dr. Dorninger served as an EMT for the Cranford First Aid Squad in Cranford, NJ. He then completed his doctorate in naturopathic medicine and his master's degree in acupuncture at Bastyr University in 2003, after which he returned to Boulder, Colorado to complete a 2-year residency in naturopathic primary care.  In 2005, Dr. Dorninger founded Roots and Branches Integrative Health Care, a clinic dedicated to “Mystery Illness” where he focuses on elucidating the underlying causes of unrelenting chronic illness.  Dr. Dorninger is not concerned with what you have as much as he is with why you have it. He has dedicated his professional life to a deeper understanding of differential diagnosis based in the tradition of “Remove Obstacles to Cure.”  After the 2013 floods in Boulder, Dr. Dorninger noticed that patients who were initially getting better suddenly regressed. Moreover, patients he was struggling to diagnose got worse.  Fortunately, he was guided to the honest, data-driven, peer-reviewed, reproducible published work of Dr. Ritchie Shoemaker on mold and biotoxin illness. In 2014 Dr. Dorninger signed on for his Shoemaker certification training.  Dr. Dorninger is a certified Shoemaker practitioner and remains true to the Shoemaker protocol.  According to Dorninger, diagnosing underlying causes and facilitating accurate treatment plans is a grind. At Roots and Branches, we embrace the grind.  In addition to private practice, Dr. Dorninger teaches functional medicine for Apex Energetics, practices Jiu-Jitsu, skis, and enjoys watching his biotoxin-genetically susceptible kids thrive post chronic inflammatory response syndrome (CIRS) treatment. Key Takeaways: What are the root causes of CIRS? What is the difference between endotoxins and lipopolysaccharides? Do Actinos or endotoxins trigger MCAS? Should Actino and endotoxin testing always be done along with the ERMI? Is there still a place for HLA-DR? What has GENIE revealed about patients with CIRS? How should Actino and endotoxin testing be evaluated? Are soil habitat Actinos a contributor to CIRS? What is the role of Actinos on the skin? What drainpipe maintenance should be considered to reduce exposure to Actinos? What steps should be taken to optimize our sleep location? Do air filters play a role in reducing Actinos and endotoxins? What is the role of Cholestyramine in dealing with Actinos and endotoxins? Can Actinos be found in the blood? Might there be a place for antimicrobial interventions in dealing with Actinos? Connect With My Guest:  https://DrDorninger.com Interview Date: November 29, 2023 Transcript: To review a transcript of this show, visit https://BetterHealthGuy.com/Episode193. Additional Information: To learn more, visit https://BetterHealthGuy.com. Disclaimer:  The content of this show is for informational purposes only and is not intended to diagnose, treat, or cure any illness or medical condition. Nothing in today's discussion is meant to serve as medical advice or as information to facilitate self-treatment. As always, please discuss any potential health-related decisions with your own personal medical authority. 

For more information, visit https://thecirsgroup.com We attended CIRSx 2023 and learned a ton of new info and are bringing you the highlights today! We had to split up the whole CIRSx weekend into two parts, and here is part two! For more information, visit TheCIRSGroup.com   TIME STAMPS: 00:00 Intro 00:58 Even Holy Water Supports Mold - Mike Schrantz and Bill Weber 03:13 Erythromelalgia and Peripheral Hypoperfusion - Dr. Lysander Jim 04:04 Stop Coronary Disease: Live Longer, Live Better - Dr. Bill Blanchett 05:26 Dust Mites: So Uncontroversial It's Almost Forgotten - Carl Grimes 07:03 Cyanobacterial Toxins as a Risk Factor for Neurodegeneration - Dr. Elijah Stommel 08:18 Coagulation and GENIE - Dr. James Ryan 09:24 Foundations Training with Jesse Salas and Dr. Goodman  10:51 Understanding Immune Response Genetics: Deeper Dive into HLA - Dr. Scott McMahon 11:52 Historical vs Modern Dust - Greg Weatherman 12:35 Osteopathy as an Adjunct Therapy for Those with CIRS - Dr. April Vukelic 14:42 Holy Water Pt 2 - Mike Schrantz and Bill Weber 15:10 House Hunting with CIRS - Jennifer Schrantz 17:09 Benzo Induced Neurologic Dysfunction - Dr. Christy Huff 19:21 Outro CIRSx website: https://www.cirsx.com/2023-cirsx-annual-conference Jacie is a 3.5+ year carnivore, certified nutrition coach, and carnivore recipe developer determined to share the life changing information of carnivore and CIRS to anyone who will listen. Barbara is a coach, facilitator, speaker, 2.5+ year carnivore, and a big fan of health and freedom. Together, they co-founded The CIRS Group, an online support community to help people that are struggling with their CIRS diagnosis and treatment. Catch Jacie and Barbara on Judy Cho's podcast to learn more about their health journeys and why they started The CIRS Group: When Carnivores are Affected by Mold Illness - Jacie Gregory & Barbara Williams - https://youtu.be/CR8Uj-d_fok

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.02.14.528561v1?rss=1 Authors: Swanson, M. E. V., Mrkela, M., Murray, H. C., Cao, M. C., Turner, C., Curtis, M. A., Faull, R. L. M., Walker, A. K., Scotter, E. L. Abstract: Microglia, the innate immune cells of the brain, are activated by damage or disease. In mouse models of amyotrophic lateral sclerosis (ALS), microglia shift from neurotrophic to neurotoxic states with disease progression. It remains unclear how human microglia change relative to the TAR DNA-binding protein 43 (TDP-43) aggregation that occurs in 97% of ALS cases. Here we examine spatial relationships between microglial activation and TDP-43 pathology in brain tissue from people with ALS and from a TDP-43-driven ALS mouse model. Post-mortem human brain tissue from the Neurological Foundation Human Brain Bank was obtained from 10 control and 10 ALS cases in parallel with brain tissue from a bigenic NEFH-tTA/tetO-hTDP-43deltaNLS (rNLS) mouse model of ALS at disease onset, early disease, and late disease stages. The spatiotemporal relationship between microglial activation and ALS pathology was determined by investigating microglial functional marker expression in brain regions with low and high TDP-43 burden at end-stage human disease: hippocampus and motor cortex, respectively. Sections were immunohistochemically labelled with a two-round multiplexed antibody panel against; microglial functional markers (L-ferritin, HLA-DR, CD74, CD68, and Iba1), a neuronal marker (NeuN), an astrocyte marker (GFAP), and pathological phosphorylated TDP-43 (pTDP-43). Single-cell levels of microglial functional markers were quantified using custom analysis pipelines and mapped to anatomical regions and ALS pathology. We identified a significant increase in microglial Iba1 and CD68 expression in the human ALS motor cortex, with microglial CD68 being significantly correlated with pTDP-43 pathology load. We also identified two subpopulations of microglia enriched in the ALS motor cortex that were defined by high L-ferritin expression. A similar pattern of microglial changes was observed in the rNLS mouse, with an increase first in CD68 and then in L-ferritin expression, with both occurring only after pTDP-43 inclusions were detectable. Our data strongly suggest that microglia are phagocytic at early-stage ALS but transition to a dysfunctional state at end-stage disease, and that these functional states are driven by pTDP-43 aggregation. Overall, these findings enhance our understanding of microglial phenotypes and function in ALS. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

Why You Should Listen:  In this episode, you will learn how the role of genomics in complex, chronic conditions. About My Guest: My guest for this episode is Dr. Sharon Hausman-Cohen.  Sharon Hausman-Cohen, MD is the co-founder, Chief Science Officer, and Medical Director of IntellxxDNA™.   Dr. Hausman-Choen has been using genomics for over a decade to deliver personalized medicine and help her patients achieve optimal health and wellness.   As a researcher and clinician advocate, she recognized that information inaccuracies and complexity were previously barriers to entry for many.   She saw the need for an easy-to-use, accurate, science-based genomics interpretation tool for clinicians, and she began to develop what is now IntellxxDNA™.   In addition to being a well-regarded doctor and researcher, Dr. Hausman-Cohen is a sought after speaker and educator on genomics, personalized medicine, and integrative medicine.   Dr. Hausman-Cohen received both her master's and medical degrees from Harvard Medical School.   She is board-certified in Family Medicine, a Fellow of the American Board of Integrative and Holistic Medicine, and possesses additional board certification in Integrative Medicine through the American Board of Physician Specialties.  She has been practicing full-spectrum Family Medicine and Integrative Medicine for more than 20 years. Key Takeaways: What is a gene?  What is a SNP?  What is genetics vs. genomics? Can modern diseases be the results of genetics? Are genes our destiny? Can supplements be used by bypass every potentially probelmatic SNP? What are the contributors to brain fog?  What can be done to improve mental clarity? What is the role of ApoE in Alzheimer's disease? How similar or different is autism to conditions like Lyme disease or MS or Alzheimer's? What role do genes play in our ability to detoxify environmental toxicants? What SNPs may play a role in biotoxin illness such as CIRS from Lyme disease or mold? Can the Lyme spirochete itself be an epigenetic influencer of gene expression? What role does the HLA-DR gene play in mold illness? Do genes play a role in immune modulation and autoimmunity? What is the connection between genes and MCAS? When is nitric oxide a good things vs. potentially a bad one? Are genes involved in hypermobility syndromes such as Ehlers-Danlos Syndrome? Are some more likely than others to have adverse reactions to anesthesia? Connect With My Guest: http://IntellxxDNA.com Interview Date: January 20, 2023 Transcript: To review a transcript of this show, visit https://BetterHealthGuy.com/Episode179. Additional Information: To learn more, visit https://BetterHealthGuy.com. Disclaimer:  The content of this show is for informational purposes only and is not intended to diagnose, treat, or cure any illness or medical condition. Nothing in today's discussion is meant to serve as medical advice or as information to facilitate self-treatment. As always, please discuss any potential health-related decisions with your own personal medical authority.

This month on Episode 41 of Discover CircRes, host Cynthia St. Hilaire highlights four original research articles featured in the September 30 and October 14 issues of Circulation Research. This episode also features an interview with Dr Kory Lavine and Dr Chieh-Yu Lin from Washington University St. Louis, to discuss their study, Transcriptional and Immune Landscape of Cardiac Sarcoidosis.   Article highlights:   Tian, et al. EV-Mediated Heart Brain Communication in CHF   Wleklinski, et al.  Impaired Dynamic SR Ca Buffering Causes AD-CPVT2   Masson, et al. Orai1 Inhibition as a Treatment for PAH   Li, et al. F. Prausnitzii Ameliorates Chronic Kidney Disease   Cindy St. Hilaire:        Hi, and welcome to Discover Circ Res, the podcast of the American Heart Association's journal, Circulation Research. I'm your host, Dr Cynthia St. Hilaire from the Vascular Medicine Institute at the University of Pittsburgh, and today I'm going to highlight articles from our September 30th and October 14th issues of Circulation Research.                                           I'm also going to have a chat with Dr Kory Lavine and Dr Chieh-Yu Lin from Washington University St. Louis, and we're going to discuss their study Transcriptional and Immune Landscape of Cardiac Sarcoidosis. But before I get to the interview, I'm going to highlight a few articles.   Cindy St. Hilaire: The first article I'm going to share is Extracellular Vesicles Regulate Sympathoexcitation by Nrf2 in Heart Failure. The first author of this study is Changhai Tian, and the corresponding author is Irving Zucker, and they are at University of Nebraska. After a myocardial infarction, increased oxidative stress in the heart can contribute to adverse cardiac remodeling, and ultimately, heart failure. Nrf2 is a master activator of antioxidant genes, suggesting a protective role, but studies in rats have shown its expression to be suppressed after MI, likely due to upregulation of Nrf2-targeting microRNAs. These microRNAs can also be packaged into vesicles and released from stressed heart cells.   Now, this group has shown that rats and humans with chronic heart failure have an abundance of these microRNA-containing EVs in their blood. In the rats with chronic heart failure, these extracellular vesicles were found to be taken up by neurons of the rostral ventrolateral medulla, RVLM, wherein the microRNA suppressed Nrf2 expression. The RVLM is a brain region that controls the sympathetic nervous system, and in the presence of EVs, it is ramped up by sympathetic excitation. Because such elevated sympathetic activity can induce the fight or flight response, including increased heart rate and blood pressure, this would likely worsen heart failure progression. The team, however, found that inhibiting microRNAs in the extracellular vesicles prevented Nrf2 suppression in the RVLM and sympathetic activation, suggesting the pathway could be targeted therapeutically.   Cindy St. Hilaire:        The next article I want to highlight is titled, Impaired Dynamic Sarcoplasmic Reticulum Calcium Buffering in Autosomal Dominant CPVT2. The first author of this study is Matthew Wleklinski, and the corresponding author is Bjӧrn Knollmann, and they are at Vanderbilt University.   Exercise or emotional stress can prompt the release of catecholamine hormones, which induce a fast heart rate, increased blood pressure, and other features of the fight or flight response. For people with catecholaminergic polymorphic ventricular tachycardia, or CPVT, physical activity or stress can cause potentially lethal arrhythmias. Mutations of calsequestrin-2, or CASQ2, which is a sarcoplasmic reticulum calcium-binding protein, is a major cause of CPVT, and can be recessive or dominant in nature.   For many recessive mutations, disease occurs due to loss of CASQ2 protein. This group investigated a dominant lysine to arginine mutation in this protein, and found by contrast, protein levels remain normal. In mice carrying the mutation, not only was the level of CASQ2 comparable to that in control animals, but so, too, was the protein's subcellular localization. The mutation instead interfered with CASQ2's calcium binding or buffering capability within the sarcoplasmic reticulum. The result was that upon catecholamine injection or exercise, the unbound calcium released prematurely from the sarcoplasmic reticulum, triggering spontaneous cell contractions. In uncovering this novel molecular etiology of CPVT, the work provides a basis for studying the consequences of other dominant CASQ2 mutations.   Cindy St. Hilaire:        The next article I want to highlight is from our October 14th issue of Circulation Research, and the title of the article is ORAI1 Inhibitors as Potential Treatments for Pulmonary Arterial Hypertension. The first author is Bastien Masson, and the corresponding author is Fabrice Antigny, and they're from Inserm in France. In pulmonary arterial hypertension, the arteries of the lungs become progressively obstructed, making it harder for the heart to pump blood through them, ultimately leading to right ventricular hypertrophy and heart failure. A contributing factor in the molecular pathology of pulmonary arterial hypertension is abnormal calcium handling within the pulmonary artery smooth muscle cells. Indeed, excess calcium signaling causes these cells to proliferate, migrate, and become resistant to apoptotic death, thus leading to narrowing of the vessel.   This group now identified the calcium channel ORAI1 as a major culprit behind this excess signaling. Samples of lung tissue from pulmonary arterial hypertension patients and a pulmonary arterial hypertension rat model had significantly upregulated expression of this channel compared with controls. And in patient pulmonary arterial smooth muscle cells, the high ORAI1 levels resulted in heightened calcium influx, heightened proliferation, heightened migration and reduced apoptosis. Inhibition of ORAI1 reversed these effects. Furthermore, in pulmonary hypertension model rats, ORAI1 inhibition reduced right ventricle systolic pressure and attenuated right ventricle hypertrophy when compared with untreated controls. This study indicates that ORAI1 inhibitors could be a new potential target for treating this incurable condition.   Cindy St. Hilaire:        The last article I want to share is titled Faecalibacterium Prausnitzii Attenuates CKD via Butyrate-Renal GPR43 Axis. The first author of this study is Hong-Bao Li, and the corresponding author is Tao Yang, and they are from the University of Toledo.   Progressive renal inflammation and fibrosis accompanied by hypertension are hallmarks of chronic kidney disease, which is an incurable condition affecting a significant chunk of the world's population. Studies indicate that chronic kidney disease is linked to gut dysbiosis. Specifically, depletion of lactobacillus bifidobacterium and faecalibacterium, prompting investigations into the use of probiotics. While supplements including lactobacillus and bifidobacterium have shown little effectiveness in chronic kidney disease, supplementations with F. prausnitzii have not been investigated.   Now, this group has shown in a mouse model of chronic kidney disease that oral administration of F. prausnitzii has beneficial effects on renal function, reducing renal fibrosis and inflammation. This bacterial supplementation also produced the short chain fatty acid butyrate, which was found to be at unusually low levels in the blood samples from the CKD model mice and from chronic kidney disease patients. Oral supplementation with this bacterium boosted butyrate levels in the mice, and in fact, oral administration of butyrate itself mimicked the effects of the bacteria. These findings suggest that supplementation with F. prausnitzii or, indeed, butyrate could be worth investigating as a treatment for chronic kidney disease.   Cindy St. Hilaire:        Today I have with me Dr Kory Lavine and Dr Chieh-Yu Lin from Washington University St. Louis, and we're going to talk about their paper, Transcriptional and Immune Landscape of Cardiac Sarcoidosis. This is in our September 30th issue of Circulation Research. Welcome, and thank you for taking the time to speak with me today.   Chieh-Yu Lin:             Thank you for inviting us. It's a great honor to be here today.   Kory Lavine:               Thank you.   Cindy St. Hilaire:        Really great paper, ton of data, and hopefully, we can pick some of it apart. But before we get into it, I actually want to just talk about sarcoidosis generally. I know it's a systemic inflammatory disease that has this kind of aggregation of immune cells as its culprit, and it can happen in a bunch of different organs. It's mostly in the lung, but it's also, like you're studying, in the heart. Can you just give us a little bit of background? What is sarcoidosis, and how common is cardiac sarcoidosis?   Chieh-Yu Lin:             Well, this is actually a great question, and I'll try to answer it. You actually capture one of the most important kind of features for sarcoidosis. It happens in all kind of organ system, mostly commonly in lung, in lymph nodes, but also in heart, spleen, even in brain, or even orbit, like eyes. It's really a truly multisystemic disease that has been characterized by this aggregate of macrophages, or myeloid cells, with scattered multinucleated giant cells, as the name implies, have multiple nuclear big, chunky, cells that form an aggregate. That's kind of like a pathognomonic feature for sarcoidosis, whether it's happening in lung, in the heart. When any organ system, a lot of studies has been done, but as of now, a very clear pathogenesis or mechanism has been, I would say, still pretty elusive, or still remain quite unclear, despite all the great effort has been made in this field. The other thing is that a lot of the studies actually focusing on pulmonary sarcoidosis for good reasons. Actually, that's one of the most common manifestations. For cardiac sarcoidosis, although it's only effect in probably, I would say depends on the data, 20% to 30% of the outpatient that with sarcoidosis, with or without lung involvement. It's actually carry a very significant clinical implications as of matter that the presentation of cardiac sarcoidosis can be devastating and sometimes actually fatal. Some of the study actually show that cardiac sarcoidosis actually higher, up to 80%, just because the first presentation's actually, unfortunately, sudden cardiac death. That's why Kory and I, we teamed up. I'm a cardiothoracic pathologist, so in my clinical practice I see specimens and samples from human body, from patient suffer from sarcoidosis, both in lung, lymph node, and heart. Kory is an outstanding heart failure, heart transplant cardiologist, see the other end, which is the patient care. This disease, specifically in heart, its presentation and its pathogens in heart, really attracts our attention.   Cindy St. Hilaire:        Do we know any or some of the potential causes? Why it would start, maybe in a different patient population, but also in the heart versus the lung? Do we know anything about that process?   Kory Lavine:              We know nothing about it. Sarcoid has no known etiology. There's been thoughts in the past that it may be driven by infection, the typical pathogens or autoimmune ideologies, but really, there's little data out there to support those possibilities. Right now, the field's wide open. The other challenge is we don't really have a good way to treat this disease, so a lot of the therapies available are things like steroids, which can have some effect on the disease but carry a lot of risk of complications. The other agents that we sometimes use to lower the doses of steroids, things like methotrexate and azathioprine, are only modestly effective.   These are really the motivation for Chieh-Yu and myself to pursue this. We don't really know what causes the disease, and we don't really have very good treatments. We really wanted to take the first step, that's to study the real disease, and understand what are the pathologic cell types that are present within the granuloma, which is these aggregation of immune cells that Chieh-Yu was speaking about.   Cindy St. Hilaire:        What is actually happening at the beginning of this disease? These granulomas form, and then what is the pathological progression in the heart? What goes on there?   Chieh-Yu Lin:             This is actually another great question that I will say there's not much that has been discovered because, especially in human tissue, every time we have a sample, it's actually a kind of time point. We cannot do a longitudinal study. But in general speaking, very little is known about how it's initiated because it will need to accumulate to a certain disease burden for this to have a clinical symptom sign and be manifested, and then being clinically studied. We do know that in both heart and lung after treatment of progressions, it's usually in, a general speaking, going through a phase from a more proliferative means that it's creating more granulomas, more  inflammatory cell aggregate, to a more fibrotic phase. Means that sometimes you actually see the granuloma start to disappear or dissipate, and then showing this kind of dense collagen and fibrosis. That has been commonly documented in both lung and heart sarcoidosis. The other things is that very difficult to study this disease that we do not have a great animal model, so we cannot use animal model to try to approximate or really study the disease pathogenesis. There are several animal models they try to use microbacteria or infectious agents, and these infectious agents can create morphologically similar granuloma, per se, but just like in human body. For instance, patients suffer from TB in their lung, biopsy will show this. But clinically, these are two very distinct disease entities, even though they look alike. Even in the heart, one of the conditions that we study in our paper is giant cell myocarditis, as the name implying having multinucleated giant cells granuloma. It looks really alike under microscopy for pathologists like me, but their clinical course in response to treatment is drastically different. This type of barriers and in the current limitations of our study tool makes, as Kory just said, this is really a wide open. We just know so little despite all the effort.   Cindy St. Hilaire:        Yeah. I'm guessing based on this granuloma information, to start with, the obvious question you went after is going after the immune cell populations that possibly contribute to sarcoidosis. To do this, because you have the human tissue, you went for single cell transcriptional profiling, which is a great use of the technology. But what biological sources did you use, and how did you go about choosing patient? Because the great thing about single cell is you can do just that, you can look at however many thousands of cells in one patient. But how do you make sure or check that that is broadly seen versus just a co-founding observation in that patient?   Kory Lavine:               We use explanted hearts and heart tissue from patients that underwent either heart transplantation or implementation of LVADs. It's a pretty big hunk of myocardium, and we're lucky to work with outstanding pathologists both at WashU, JU, as well as our collaborators at Duke. Between the two institutions, we're able to pull together a collection of tissues where we knew there were granulomas within that piece of tissue we analyzed. You bring up an important challenge. You need to make sure the disease and cause of the disease is present in the tissue that you're analyzing, otherwise you'll not come up with the data that really is informative.   Chieh-Yu Lin:             Kory beautifully answered the question, but I just wanted to add one little thing, and that's also why we use various different modalities. Some of them is more inside you, like the NanoString Technologies' spatial transcriptomic. You can visualize and confirm that we are studying the phenomenon that has been described for sarcoidosis, and then using multichannel immunofluorescence to validate our sequencing data, to complement such limitations of certain technology.   Cindy St. Hilaire:        Especially, I feel like with this diseased tissue that it's such a large tissue, there's so much information, it's really hard to dig in and figure out where the signal is. This was a wonderful paper for kind of highlighting, integrating all these new technologies with also just classical staining. Makes for great pictures as well. How does this cellular landscape of cardiac sarcoidosis compare to a normal heart? What'd you find?   Chieh-Yu Lin:             This is a great question. Compared to normal heart, we have been talking about this accumulation of macrophages with scattered multinucleated giant cells. For the similar landscape, first and foremost, you do not see those type of accumulations in brain microscopy or by myeloid markers in the heart. Although, indeed, in even normal heart tissue we have rest and macrophages. It just doesn't form such morphological alterations. But then we dive deep into it, and then we found that from a different cell type perspective, we realized that the granuloma is composed by several different type of inflammatory cells, with most of the T cells and NKT cells kind of adding periphery. The myeloid cells, including the multinucleated giant cells also, are kind of in the center of the granuloma of the sarcoidosis. Then, we further dive in and realize that there are at least six different subtype of myeloid cells that is contributing to the formation of this very eye-catching distinctive granular malformations, and to just never feel first off and foremost, of course, is those multinucleated giant cells that is really distinct, even on the line microscopy] routine change stand.   And then we have a typical monocyte that's more like a precursor being recently recruited to the heart, and we finally sent the other four different type of myeloid cell that carry different markers, and then improving the resident macrophages. Especially for me as a pathologist, I'm using my eye and looking at stand every day, is actually these six type of cells, myeloid cells, actually form a very beautiful special kind of distribution with the connections or special arrangement with all different type, kind of like multinucleated giant cell in the middle, flanked by HLA-DR positive epithelioid macrophages, kind of scatter, and then with dendritic cells and a typical monocyte at the peripheral, and then resident macrophage kind of like in the mix of the seas of granuloma information. All these are distinct from normal heart tissues that does carry a certain amount of macrophages, but just don't form this orchestrated architectural distinct structure that's composed of this very complicated landscape.   Cindy St. Hilaire:        Those images, I think it was figure six, it's just gorgeous to look at, the model you made. One of the questions I was thinking is there must be a significance between these cells that are on the periphery and those that are in the center of this granuloma. Do you have an idea or can we speculate as to are some more cause and some more consequence of the granuloma? Were you able to capture any more information about maybe the initiating steps of these from your study?   Kory Lavine:              That's a great question, and a question the field has had for a long time. Now, we know there's different populations of cells. The single cell data allows us to understand what are the transcriptional differences and distinctions between them to gain some insights. One thing that we do know from the field is that disease activity correlates with mTOR activity within these granulomas. We took advantage of phospho-S6 kinase staining as a downstream marker of mTOR activity, and Ki-67 is a marker of self proliferation.   Which of these populations within the granuloma might be most active with respect to mTOR and respect to proliferation? If you ask most people in the field, they would jump up and say, "It's the giant cell in the middle." We found that that's not actually the case at all. It's the macrophages that surround the giant cell, the ones that are HLA-DR positive, the epithelioid macrophages, and the ones that are SYLT-3 positive that are scattered around them. That's really interesting and could make a lot of sense, and leads to hypothesis that perhaps activation mTOR signaling within certain parts of the granuloma might be sufficient to set up the rest of the architecture. That's something that we can explore in animal models, and are doing so to try to create a cause and effect relationship. Cindy St. Hilaire:        Yeah, and I was actually thinking about this, too, in relation to kind of the resident macrophages versus infiltrating macrophages or even just infiltrating immune cells. Do you know the original source of the cells that make up the granuloma? Is it mostly resident immune, or are they recruited in?   Kory Lavine:               We can make predictions from the single cell data where you can use trajectory analysis to make strong predictions about what the origin of different populations might be. What those analyses predicted is that the giant cells and the cells that surround the giant cells, the HLA-DR positive and SYLT-3 positive macrophages, come from monocytes. That's the prediction, and, of course, resident macrophages do not. However, that prediction has to be tested, and that's the beauty and importance of developing animal models. The wonderful thing today is we now have genetic tools to do that. We can ask that question.   Cindy St. Hilaire:        I don't know. Maybe you don't want to spoil the lead of the next paper, but what kind of mouse model are you thinking about trying?   Kory Lavine:               Yeah. First of all, let me talk about the tools that are available, because they're published in Circulation Research, of course. We have a nice tool to specifically mark, track and delete in tissue resident macrophages using a CX3CR1 ERT pre-mouse, and taking advantage of the concept that tissue macrophages don't turn over from monocytes and turn over from themselves. We can give tamoxifen to label all monocytes macrophages in Dcs with that CRE, and then wait a period of time where only the resident macrophages remain labeled. We can use that trick to modulate mTOR signaling as a first step, and ask whether mTOR signaling is required in that population. We've now developed a new genetic tool to do the same thing in just recruited macrophages.   Cindy St. Hilaire:        What was the most challenging aspect of this study? There's a lot of moving parts. I'm sure probably the data analysis alone is challenging, but what would you say is the most challenging?   Kory Lavine:               I think you alluded to this early on, but the most challenging thing is collecting the right tissues to analyze, and that's not a small feat or a small effort here. All the technologies are a lot of fun, and everything works so well today compared to many years ago when we trained, so it's an exciting time to do science. The most challenging and time-consuming component was assembling a group of tissues that we could do single-cell sequencing on between our group and our colleagues at Duke, and then creating validation cohorts that we did across several different institutions, including our own as well as Stanford. That team effort in building that team is the most important, challenging, and honestly, enjoyable part of this.   Chieh-Yu Lin:             I cannot agree more what Kory just said. I think that that's the challenging and the fun part, and that we're very fortunate to really have a great team to tackle this questions in multiple from multiple institute. I just want to add one more thing that, particularly for me as a cardiopathologist, one of the hardest things is I've known how to look or diagnose sarcoidosis for years, but seeing the data emerging that is so complicated and then beyond my reliable eyes in understanding, it's kind of mentally very challenging but very fun to really open and broaden the vision. It's not just how it looks like just giant cells in macrophages.   Cindy St. Hilaire:        What do you think about in terms of diagnostics or even potential therapies? How do you think this data that you have now can be leveraged towards those objectives, whether it's screening for new cell types that are really key to this granuloma formation versus therapeutically targeting them?   Kory Lavine:              This study opens new doors, and right now, diagnosis of sarcoids islimited by trying to biopsy, which, in the heart, is limited by sample bias. You certainly can biopsy the wrong area because you don't know whether a granuloma is in the area or not. We do do some cardiac and other imaging studies like FDG-PET scans, which are helpful but are not perfect, and each of them has their individual limitations. One of the beauties of our study is it identifies new markers of macrophage populations that live within the granuloma, many of which are unique to this disease.   That suggests that there's maybe an opportunity to develop imaging tracers that can identify those populations more specifically than our current PET imaging studies do, which rely simply on glucose uptake. It also opens up the possibility that we may able to take blood samples and identify some of these cell types within the blood, and have more simple testing for our patients. I think in terms of therapy, you alluded to it earlier, these concepts about mTOR signaling, that could be a new therapeutic avenue that needs to be rigorously explored in preclinical models. We're lucky already to have very good mTOR inhibitors available in clinical practice today.   Cindy St. Hilaire:        Obviously, opening new doors is amazing because it's more information, but often a good study leads to even more questions to be asked. What question, or maybe what questions, are you guys going to go after next?   Chieh-Yu Lin:             Well, that list is very long, and then that's actually the exciting thing about doing this research. There's no bad questions, in some sense. All the way from diagnosis, management, monitoring, therapeutic, how we predict where the patient can respond, that's the whole clinical side. Even the basic science side, we still haven't really answered the question, although our data suggests where that multinucleated giant cells coming from. It's very eye catching. How do they form, even though our data suggests it's from the recruited macrophages. But that's still a long way from the recruited macrophage,  monocyte to that gigantic bag of nuclei in the very fluffy cytoplasm.   And then, how the granuloma, as we discussed earlier in this discussion, really initially from a relatively normal background myocardium to form this disease process. There are just so many questions that we can ask. There are, of course, several fronts that we would like to focus on. Kory already nicely listed some of them. First and foremost is actually to establish animal model to enable us to do more details in mechanistic studies, because human tissue, as good as it is, it's kind of like a snapshot, just one time point, and it really limits our ability to test our hypothesis. Animal model, certainly, is one of the major directions that we are going forward, but also the other side, like more clinical science also to develop novel noninvasive methodologies to diagnose and to hopefully monitor this patient population in a better way.       Cindy St. Hilaire:        Well, it's beautiful work. I was actually reading this paper this weekend at a brunch place just next door to my house, and the guy sitting next to me happened to see over my shoulder the title and said that his father had passed away from it. This is hopefully going to help lots of people in the future, and really help to make the models that we need to ask, "What's happening in this disease?" Thank you so much for taking the time to speak with me, and congratulations on what seems to be a landmark study in understanding what's going on in this disease.   Chieh-Yu Lin:             Thank you so much. It's a pleasure.   Cindy St. Hilaire:        That's it for our highlights from the September 30th and October 14th issues of Circulation Research. Thank you so much for listening. Please check out the Circ Res Facebook page, and follow us on Twitter and Instagram with the handle @CircRes, and hashtag Discover Circ Res. Thank you so much to our guests, Dr Kory Lavine and Dr Chieh-Yu Lin from Washington University St. Louis. This podcast is produced by Ashara Retniyaka, edited by Melissa Stoner, and supported by the editorial team of Circulation Research. Some of the copy texts for highlighted articles was provided by Ruth Williams. I'm your host, Dr Cynthia St. Hilaire, and this is Discover Circ Res, your on-the-go source for the most exciting discoveries in basic cardiovascular research. This program is copyright of the American Heart Association, 2022. The opinions expressed by speakers in this podcast are their own, and not necessarily those of the editors of the American Heart Association. For more information, please visit ahajournals.org.  

Emergencies happen in hematology and oncology. This is a fact. But how do we manage these emergencies? Look no further. In this episode, we'll talk about one of the key hematologic malignancies that you'll encounter as a fellow, one that requires immediate action to reduce mortality: acute promyelocytic leukemia (APL or APML)- Acute Promyelocytic leukemia (APL or APML):**Stay tuned for our upcoming “part two” and “chemotherapy basics” episodes for more information on non-acute management of this disease**APL is a true hematologic emergency! Although this is a very curable form of leukemia, it is associated with high rates of severe DIC and high mortality in the period immediately following diagnosis***Untreated, can see pulmonary or cerebrovascular hemorrhage in up to 40% of patients***10-20% incidence of hemorrhage-related mortality in the initial period***Statistically significant increase in mortality at 30 days with just a 12-hour delay in initial hematologist consultation- Disease basics:** Rare subtype of AML(

Shannon Martin is an Integrative Health Coach and Mineral Balancing Practitioner. Her interest in holistic health started when she realized that environmental factors were contributing to her gut health issues. Shortly after finishing her coaching certification, Shannon and her family started to develop symptoms due to toxic mold in their home. This experience deepened her knowledge on the influence the environment and nutrient deficiencies have on our ability to heal. Shannon now educates about toxins and their impact on our bodies. She offers one-on-one coaching and helps people safely detox toxic mold, heavy metals, and chemicals. Her holistic approach includes replenishing nutritional deficiencies so that the body can heal. Full shownotes for this episode here. SHOWNOTES: 07:12 — Mold and Its Effects on Our Health When to start investigating Can a brand new house have mold? What is the HLA-DR gene? 18:43 — Modern State of Healthcare How generations of people are used as test subjects How many chemicals are actually tested? 21:07 — The Symptoms of Mold Exposure The variety of mold exposure symptoms Asking “where” and “when” questions to determine the cause of your symptoms 25:36 — Toxins in the Modern World Why toxins accumulate in the brain How toxins can prevent you from loosing weight How to not get overwhelmed if you are just starting on your health journey Number one ingredient to look out for in your beauty products 35:43 — The Dangers of Fluoride The connection between high fluoride levels and bone fractures How fluoride actually weakens your teeth and bones A visible sign you have too much fluoride The effect of fluoride on the pineal gland How fluoride got into our water: the story about a man who convinced the whole world that fluoride is necessary for dental health 43:16 — Toxic Heavy Metals Mercury in dental amalgam fillings The truth about toxic heavy metals How to prepare for an amalgam filling removal 46:55 — The Importance of Minerals Over 90% of people are deficient in minerals Where are the minerals in our food coming from? What are preferred minerals? How to balance the minerals in your body What makes us absorb more toxic minerals? Resources: Website: www.bewellandgreen.com Instagram: @toxsick_podcast ••• Love the show? Here are the ways you can support it: 1. SUBSCRIBE on Apple Podcasts • Google Podcasts • Spotify • Stitcher 2. LEAVE A REVIEW on Apple Podcasts 3. SHARE with friends, family members, or anyone who you think will love the show, too! Follow on INSTAGRAM for more better life inspiration: @betterwaytodolife

Today's episode is dedicated to Critical Illness In Children With Hematopoietic Stem Cell Transplants. We are delighted to be joined by Dr. Muna Qayed, Associate Professor of Pediatrics Emory University School of Medicine , Atlanta, GA. She is also the Director of the Blood and Marrow Transplant Program at the Aflac Cancer and Blood Disorders Center at Children's Healthcare of Atlanta. Our Case: A 10 year old female with refractory high-risk ALL s/p mismatched unrelated donor transplantation T+13 days presents as a transfer to the PICU with abdominal distention, worsening jaundice, and escalating nasal cannula requirements. The patient's post-transplant course was complicated by gram-negative bacteremia requiring fluid resuscitation. A CXR upon transfer to the PICU is notable for bilateral airspace disease, a right sided pleural effusion, and hypoexpanded lung fields. The patient is promptly intubated, sedated and started on renal replacement therapy. Echo labs, and further imaging are pending. What are the classic pediatric indications for BMT? Autologous BMT (where donor cells are from the patient/recipient) is used as consolidation in some solid tumors such as High risk neuroblastoma, brain tumors like medulloblastoma, and germ cell tumors, and are a standard treatment approach in relapsed Hodgkin lymphoma Allogeneic BMT-where in the donor cells are derived from another individual are typically used for hematologic malignancies. ALL and AML are most common pediatric indications. Also allogeneic BMT are used for wide spectrum of nonmalignant hematology conditions such as hemoglobinopathies ( Sickle cell disease, Thalassemia), and severe aplastic anemia, and inherited bone marrow failure syndromes, as well as some metabolic disorders and immune-deficiency disorders such as SCID, HLH and other primary immune regulatory disorders. The sources of graft in BMT? Stem cells (which give rise to different types of blood cells - red cells, white cells and platelets are derived from the bone marrow. Thus the overall process is known as Bone Marrow Transplantation. Stem cells can be also derived from peripheral blood - when the donor is treated with granulocyte colony stimulating factor or G-CSF. There are some key advantages here, which include the ability to collect a much higher stem cell dose, with faster hematopoietic recovery. However the downside is a higher T cell content of the graft with subsequent increased risk of graft versus host disease. Umbilical cord blood is also used as a source of stem cells. Mega doses of stem cells are used to overcome histocompatibility barriers of mismatched transplantation. Majority of T cells have to be removed from donor pool to prevent severe GVHD., Increase risk of infection and relapse of patients original disease. Explain the human leucocyte antigen (HLA) and its role in BMT? The Major Histocompatibility complex (MHC) system known as the human leukocyte antigen (HLA) in humans is located on the short arm of chromosome 6 and contains the most polymorphic gene cluster of the entire human genome. The HLA consists of regions designated as "classes". Class I and class II are relevant to stem cell transplant. The main function of HLA class I gene products (HLA-A, -B, and -C) is to present endogenous peptides to responding CD8+ T Cells, HLA class I antigens are expressed on all nucleated cells and platelets. While the class II coded molecules HLA-DR, -DP, and –DQ have restricted expression and process exogenous peptides for presentation to CD4+ helper T Cells, and are expressed on antigen presenting cells. HLA-A, HLA-B, HLA-C and HLA-DR are traditionally the loci critical for matching for stem cell donor. In addition to deciding on the source of the graft, we have to make decisions on who the donor will be. If a matched sibling donor is not available (or in some inherited conditions that may not be an option as a donor), then matched unrelated donors or matched cord blood...

Thank you for joining us for our 2nd Cabral HouseCall of the weekend! I’m looking forward to sharing with you some of our community’s questions that have come in over the past few weeks… Let’s get started!    Anonymous: Hi Doc, First thing I’d like to do is thank you. Great podcast, you’re helping a lot of people with all the information shared. Keep it up. I have a mold question in regards to genetics; The most popular mold physician says testing for specific HLA-DR genes is necessary to determine if the body is creating the antibodies to go after the molds in the body. I have what he calls the “dreaded gene” that is found in only 1% of population, meaning multiple sucestibility to mold/lyme/etc as my body apparently doesn’t create the antibodies against them. On top of this, I have a MTHFR mutation. So my question is wether this HLA-DR gene testing matters or not? Do the MTHFR/HLA-DR genetic tests all mean the same thing? There is much debate as to the credibility behind this theory so I was hoping to hear your thoughts on it. Thanks! Jenett: Hi Dr. Cabral, My friend was diagnosed with ocular pressure due to an imbalance of eye fluid production. He as told it can lead to glaucoma and blindness. Is there a way to balance this naturally? Thank you! Jenett Marissa: Dr. Cabral, I really appreciate you and all your time spent on sharing your years of reading and knowledge with our community. I found you by my research on Ayurveda and since finding you have listened to almost every podcast you have done. I feel like I have gotten my own "certification" in wellness by having all of this information you have shared. Thank you!! You have helped me, my friends, and my family live better. I have done a DCD, CBO protocol/finisher, heavy metal detox and use your shake, adrenal soothe, CBD, and other products daily. I am a 40-year-old vata body type and enjoy the gym. I like going in the morning (5am) and usually do it fasted. I don’t want any food, but some days I have a ½ banana for being vata (per your recommendation.) I have noticed over the years that I cannot do cardio to almost any degree without my muscles getting tight after and I used to get sleepy post the workout (approx. 2-3 hours.) I tried electrolytes during the gym, but it doesn’t help. My HTMA markers are normal. I enjoy being active, but have learned to not push it for the past 2 years, thinking that my body is too stressed. I now stick to mainly body weight exercises and no cardio, but I like getting better oxygenation with some increase in heart rate. Also, my morning temperature does not get over 97.6 degrees most days, only on the 2nd half of my cycle. My most recent Dutch hormone test showed my rising morning cortisol had been slightly low and my morning cortisol slightly high, with normal ranges the rest of the day. I test my HRV with my Oura ring and struggle to get that over the 40’s. My TSH in the past was between 5-9 and now is lowered to 2-3 with the help of Armour Thyroid at its lowest dosage. Why can’t my temperature, HRV go up? If you met me you wouldn’t think I had a low energy. I am lean and not overweight. I want to get all metabolism markers higher. Any guidance would help!  Mrs M: Dear Dr Cabral, I have searched but found no results regarding bunions (hallux valgus.) Can you recommend a natural treatment that does not involve surgery? I did a Google search and found all kinds of splints / straps / bandages and toe separators. I would very much appreciate if you could share any ideas on how to remedy this situation. I have only just begun to experience pain, I do not have a large growth and it’s only on my one foot. The big toe only points towards my other toes slightly. My mother and grandmother had it quite badly (my mother had hers removed surgically.) I have large, wide feet (European Size 43) which makes finding comfortable, well-fitting shoes for work (office job) nearly impossible. I’m also a runner but do not yet feel much pain when running (yet?) Would very much like to avoid surgery if possible. Thank you in advance for your help and feedback, it is very much appreciated. Mrs M. Katie: Hi Dr. Cabral, This is the second time I've written in to the Cabral concept and instead of listing symptoms, I'd like to ask you a more over-arching health question. I'm in my late 20s and have been struggling with physical and mental health issues for most of my adult life. As a young woman I made most of my mistakes early on because I was ignorant to the health care system and honestly wasn't a great advocate for myself. I allowed health professionals to diagnose and prescribe medication without questioning any of their methods. I thought doctors were altruists who cared for my well being and ultimately wanted to heal their patients. I was very wrong. I have a deep sense of distrust of doctors and although I see more healing in the natural medicine and holistic industry, I still find myself questioning whether the treatments will work. It's a different business than the traditional health care industry, but a business none the less. How do you, I, or anyone for that matter, sift through all the information out in the world today, and how do we know who to trust? Sometimes I wish life came with an instruction manual. I'm feeling defeated and would love your insight. Thanks for always spreading your knowledge and positivity.   Thank you for tuning into this weekend’s Cabral HouseCalls and be sure to check back tomorrow for our Mindset & Motivation Monday show to get your week started off right! - - - Show Notes & Resources:  http://StephenCabral.com/1213 - - - Dr. Cabral's New Book, The Rain Barrel Effect https://amzn.to/2H0W7Ge - - - Join the Community & Get Your Questions Answered: http://CabralSupportGroup.com - - -   Dr. Cabral’s Most Popular Supplements: > “The Dr. Cabral Daily Protocol” (This is what Dr. Cabral does every day!) - - - > Dr. Cabral Detox  (The fastest way to get well, lose weight, and feel great!) - - - > Daily Nutritional Support Shake  (#1 “All-in-One recommendation in my practice) - - - > Daily Fruit & Vegetables Blend  (22 organic fruit & vegetables “greens powder”) - - - > CBD Oil  (Full-spectrum, 3rd part-tested & organically grown) - - - > Candida/Bacterial Overgrowth, Leaky Gut, Parasite & Speciality Supplement Packages - - - > See All Supplements: https://equilibriumnutrition.com/collections/supplements  - - -   Dr. Cabral’s Most Popular At-Home Lab Tests: > Hair Tissue Mineral Analysis (Test for mineral imbalances & heavy metal toxicity) - - - > Organic Acids Test (Test for 75 biomarkers including yeast & bacterial gut overgrowth, as well as vitamin levels) - - - > Thyroid + Adrenal + Hormone Test  (Discover your complete thyroid, adrenal, hormone, vitamin D & insulin levels) - - - > Adrenal + Hormone Test (Run your adrenal & hormone levels) - - - > Food Sensitivity Test (Find out your hidden food sensitivities) - - - > Omega-3 Test (Discover your levels of inflammation related to your omega-6 to omega-3 levels) - - - > Stool Test (Use this test to uncover any bacterial, h. 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Why You Should Listen: In this episode, you will learn about healing your body from toxic illnesses such as Mold Toxicity, Lyme Disease, Multiple Chemical Sensitivities, and Chronic Environmental Illness. About My Guest: My guest for this episode is Dr. Neil Nathan. Neil Nathan, MD was introduced with a more formal summary of his bio in podcast episode #19. For this podcast, I want to be more informal and just say how much I value and respect Dr. Nathan and the work that he does to help his patients. Dr. Nathan has been a mentor to me over the years, and I am always grateful when there are more opportunities to learn from him. I appreciate that Dr. Nathan has never accepted the toolbox that he has available and that he has continued to look for new and better options to help his population of chronically ill patients. While his toolbox is quite large, he’s still looking, still searching, still open to new and different approaches. Until we have all the answers, this is the type of doctor I think most of us would want. In his new book “Toxic: Heal Your Body from Mold Toxicity, Lyme Disease, Multiple Chemical Sensitivities, and Chronic Environmental Illness”, Dr. Nathan shares his wisdom, experience, and toolbox with all of us. It is profoundly enlightening. Key Takeaways: - What is the difference between toxicity and sensitivity? - What is a binder? How are absorption and adsorption different? - How does one determine which binders to use? - Can binder effectiveness be maximized with a focus on the liver and gallbladder? - How are intracellular toxins detoxified? - How important are HLA-DR genetics in terms of being a predictor of illness and a predictor of treatment outcome? - What screening tests might be used to evaluate an environment for the possibility of mold? - How are urine mycotoxin tests helpful? - How can mold from a water-damaged building vs. mold from food consumption be differentiated in urine mycotoxin tests? - How often is colonization a consideration in recovering from mold illness? - Does treating MARCoNS make a positive shift for patients clinically? - How do Bartonella and Babesia differ in their symptom presentation? - What is provocation testing? - Can Bartonella be treated without antibiotics? - What is Mast Cell Activation Syndrome, and how is it treated? - How might various body systems be rebooted? - What is Polyvagal Theory, and what role does it play in recovery? Connect With My Guest: http://neilnathanmd.com Related Resources: Book:"Toxic: Heal Your Body from Mold Toxicity, Lyme Disease, Multiple Chemical Sensitivities, and Chronic Environmental Illness"  International Society for Environmentally Acquired Illness Interview Date: October 22, 2018 Additional Information: To learn more, visit http://BetterHealthGuy.com. Disclaimer: The content of this show is for informational purposes only and is not intended to diagnose, treat, or cure any illness or medical condition. Nothing in today's discussion is meant to serve as medical advice or as information to facilitate self-treatment. As always, please discuss any potential health-related decisions with your own personal medical authority.

Distinguir lo propio de lo ajeno, y lo normal de lo aberrante, no es tarea fácil. De hecho, una parte importantísima de nuestra energía se consume en las operaciones del sistema inmune, el encargado de esta tarea. Para el efecto, es vital el complejo mayor de histocompatibilidad, o CMH (MHC en inglés) y su expresión en la superficie de nuestras células, a saber, las moléculas del sistema de antígenos leucocitarios o HLA, por sus siglas en inglés. Conocer a profundidad este tema nos permitirá comprender su implicación en enfermedades alérgicas, autoinmunes, cáncer y muchas más. Existen, básicamente, dos grupos de moléculas del HLA: las HLA-A, HLA-B y HLA-C, pertenecientes a la clase I; y las HLA-DP, HLA-DQ y HLA-DR, de la clase II. Las primeras, presentes en la gran mayoría de células, presentan antígenos provenientes de péptidos de síntesis endógena, que han sido fraccionados por el proteasoma, y las segundas, más característicamente halladas en células presentadoras de antígeno profesionales, albergan péptidos que vinieron de proteínas captadas del exterior de la célula, digeridas en el complejo de vesículas de los endosomas y lisosomas. Las moléculas del CMH clase I presentan sus péptidos a los linfocitos T CD8+, citotóxicos, y son el blanco de estas últimas en caso de presentar algo "indebido", por ser reconocido como ajeno o aberrante. Por otra parte, las moléculas del CMH clase II son responsables de presentar los antígenos a linfocitos T colaboradores, es decir, CD4+. Revisaremos su estructura, su síntesis, ensamblaje y transporte, y las excepciones a la regla que hemos relatado en párrafos anteriores. Revisaremos la sorprendente (y descubriremos por qué no es tan soprendente) forma en que con poquísimos aminoácidos, este complejo de moléculas de superficie puede ayudar a distinguir las proteínas propias y normales de las que no lo son, y revisaremos el fantástico rol que cumplen en esta ecuación los linfocitos NK o natural killers, las células del sistema inmunitario "asesinas por naturaleza". ¿Quieres enterarte semana a semana de las nuevas publicaciones, y acceder a contenido exclusivo? Únete a la lista de correo de Leucocitos isotópicos. Para suscribirte al Podcast de Medicina, estas son las opciones más recomendadas: Apple Podcasts Google Podcasts Si prefieres explorar más alternativas, haz clic aquí. ¿Te gustó el episodio? Seguro disfrutarás este también: El sistema inmunitario y su dualidad (027) Además, puedes acceder a la lista curada y actualizada de los episodios con mayor aceptación. Este show es para ti. Puedes apoyarlo entrando a iTunes y dejando allí una calificación positiva. Encuentra las notas de este episodio y dirige a tus amigos a isotopicos.com/020 El objetivo de Leucocitos isotópicos es entretenerte mientras complementas lo que recibes en tu Escuela o Facultad de Medicina. Soy Médico Internista, y comprendo lo demandante que puede ser nuestra Carrera. Por eso decidí crear el Podcast como un curso de Medicina ameno y sin una estructura rígida, que despierte tu interés y curiosidad por esta maravillosa Ciencia. Nunca reemplazará a la Universidad, ni a los libros, pero cumplirá con la misión que lo fundamenta: Ser el lugar donde descansamos de leer, sin dejar de aprender. No olvides que la mejor manera de ayudar a que el proyecto crezca, es contarle a todos de él. ¡Gracias por compartir este episodio con alguien!

Why You Should Listen: In this episode, you will learn about the various factors that may be roadblocks to recovering from Lyme disease including mold, parasites, dental issues, and more. About My Guest: Dr Raj Patel has extensive experience with, various natural therapies including nutritional medicine, homeopathy, herbs and mind-body medicine. Integrating these diverse therapies with allopathic medicine has enabled Dr. Patel to offer a highly refined approach to healthcare that produces lasting results. Since 1993, Dr. Patel has specialized in helping patients struggling with "Chronic Fatigue" (CFIDS), Candidiasis, food sensitivities, thyroid and other hormonal imbalances, chronic infections, and heavy metals. Over the years, Dr. Patel has specialized in treating patients with Autistim Spectrum Disorders (ASD) as well as Lyme and other vector-borne diseases. He began working with autistic children in 1999 and to date has helped over 500 children on their road to recovery. Another area that Dr. Patel specializes in is treating Lyme disease. It is well documented that patients with chronic Lyme disease also suffer from Candidiasis, food sensitivities, accumulation of heavy metals, and hormonal imbalances. With his background in treating these conditions, Dr. Patel has been able to create a comprehensive program for treating those struggling with chronic Lyme disease. He has found incredible success in treating individuals with chronic Lyme disease by combining antimicrobial treatment with heavy metal detoxification, food allergy desensitization and hormonal support. Dr. Patel is an active member of the International Lyme and Associated Diseases Society (ILADS). He has also completed advanced training in pediatric Lyme disease. Dr. Patel first began looking into Dr. Shoemaker’s incredible work on chronic inflammation resulting from exposure to water damaged buildings (WDB) in 2010. He found that many of his “Lyme patients” who had experienced only partial improvement, turned out to be simultaneously struggling with toxic exposure to water-damaged buildings. When they began responding dramatically to simply getting out of their home and taking cholestyramine, Dr. Patel’s interest was piqued to learn more about Dr. Shoemaker’s treatment approach to this illness. Dr. Patel today integrates the findings from Dr. Shoemaker as well as other doctors working with mold illness to see patients making impressive recoveries from this debilitating illness. Key Takeaways: - When mold and Lyme are both factors, what is the proper treatment order? - What is the impact of HLA-DR in recovery? - What do elevated C4a and TGFb1 mean? - Is an ERMI helpful for testing your living environment? - How is MARCoNS approached? - Does VIP help? - How is Lyme treatment approached? - Is the goal to eradicate every microbe? Is that required to get well? - Are parasites an issue in chronic Lyme disease? - What works for detoxification? - What is UVB therapy and how is it used? - How can ozone be helpful? - What is Biomagnetism and when might it be helpful? - Does mental/emotional health matter? - What diet is most helpful? Connect With My Guest: http://www.drrajpatel.net Interview Date: May 9, 2017 Disclaimer: The content of this show is for informational purposes only and is not intended to diagnose, treat, or cure any illness or medical condition. Nothing in today's discussion is meant to serve as medical advice or as information to facilitate self-treatment. As always, please discuss any potential health-related decisions with your own personal medical authority.

Beim kolorektalen Karzinom treten Biomarker zunehmend in den Fokus. Ziel muss daher die Etablierung von effektiven Markern sein, um den Patienten eine möglichst effektive, auf das Individuum zugeschnittene Diagnostik und Therapie anbieten zu können. In unserer Studie zum kolorektalen Karzinom sind nach Ausschlusskriterien n=108 Patienten mit Primärtumoren und n=59 Patienten mit Lebermetastasen vom KRK in die statistische Analyse eingegangen. Aus postoperativ entnommenem, N2-schockgefrorenem Gewebe wurde ein Biomarker-Profil aus molekulargenetischen und immun-histologischen Markern erstellt. Genetische Mutationen auf dem KRas- und BRaf-Gen wurden mittels Mikrodissektion/PCR/Pyrosequenzierung detektiert und mittels immunhistochemischer Färbungen die Moleküle EGF-R, Her2/neu, IGF1-R, c-Met, CD44v6, Ki67, CD45 und HLA-DR analysiert. Die statistische Analyse erfolgte univariat mittels Chi-Quadrat-Test, T-Test, Mann-Whitney-U-Test und Kruskal-Wallis-Test. Überlebensanalysen erfolgten mittels Kaplan-Meier-Schätzer und Cox-Regressionsanalyse. Hierbei ergaben sich in der univariaten Analyse zwischen den einzelnen Parametern der Primärtumoren folgende statistisch signifikante Korrelationen: Eine Mutation/der Wildtyp im KRas-Gen korrelierte mit dem L-Stadium, einer Mutation/der Wildtyp im BRaf-Gen und IGF1-R (p=0,007; 0,003; 0,034). Der um die Mutation G13D erweiterte KRas Wild-typ bzw. mutiert (ohne G13D) korrelierte hierunter mit dem L-Stadium, der Histologie, sowie ebenfalls mit BRaf (p=0,038; 0,039; 0,011). Eine Mutation im BRaf-Gen (Exon 15) war mit dem Geschlecht, der Lokalisation und dem Grading des Primärtumors signifikant verbunden (p=0,042; 0,003; 0,002). Der EGF-R korrelierte mit der Lokalisation, dem Grading, dem L-Stadium und CD44v6-Positivität (p=0,031, 0,020, 0,006, 0,021). Bei Her2/neu fand sich für das Geschlecht, die Lokalisation, CD45 und für HLA-DR eine Ver-knüpfung (p=0,005, 0,021, 0,032, 0,006). Zudem konnte ein Zusammenhang zwischen Her2/neu (Score) und dem Geschlecht (p=0,009) sowie der Tumorlokalisation (p=0,010) nachgewiesen werden. Für c-Met und IGF1-R (p=0,021) und für IGF1-R mit der Lokalisation des Primarius (p=0,027) bestand eine positive Korrelation. Des Weiteren korrelierten CD45 mit HLA-DR (p=0,046) und Ki67 mit dem Alter der Patienten (p=0,015). Hinsichtlich der Lebermetastasen konnte eine Verbindung von EGF-R mit der Histologie, mit IGF1-R und c-Met (p=0,046; 0,004; 0,007) nachgewiesen werden. Zudem konnte für c-Met und der Tumor-größe, dem löslichen präoperativen Tumormarker CA19-9 und CD44v6 (p=0,004; 0,003; 0,044) eine positive Korrelation nachgewiesen werden. HLA-DR und der BMI wiesen einen statistisch signifikanten Wert (p=0,022) auf und für Ki67 und CD44v6 bestand ebenfalls eine signifikante Verbindung (p=0,007). Ki67 und das Grading (p=0,017) sowie Her2/neu (Score) und die Histologie der Lebermetastasen (p=0,018) wiesen ebenfalls Signifikanz auf. Für das Gesamtüberleben im Kollektiv der Primärtumoren konnte im Kaplan-Meier-Schätzer ein Zusammenhang von N-Stadium (p

Zystische Fibrose ist die häufigste vererbbare letale Erkrankung in Europa, die trotz Fortschritten in Diagnostik und Therapie weiterhin mit einer verkürzten Lebenserwartung einhergeht. Einer der Hauptgründe verfrühter Sterblichkeit betroffener Patienten sind persistierende pulmonale Infektionen mit Pseudomonas aeruginosa. Der Keim bedient sich des Quorum Sensing (QS), eines interbakterielles Kommunikationssystems, um die Ausbildung von Virulenzfaktoren zu regulieren und die Immunantwort des Patienten zu beeinflussen. In dieser Arbeit wurde die Auswirkung des P. aeruginosa QS Moleküls 3oxoC12-HSL auf die Reifung humaner Dendritischer Zellen (DZ) untersucht. DZ vermitteln als professionelle Antigen-präsentierende Zellen zwischen angeborenem und erworbenem Immunsystem. Eine Infektion wurde simuliert, indem humane DZ mit Lipopolysaccharid oder Zytokin-Cocktail aktiviert wurden. Anschliessend wurde die Expression von Maturations- und Migrationsmarkern sowie Zytokinsekretion in Anwesenheit von 3oxoC12-HSL untersucht. Bei LPS-stimulierten DZ kam es in Anwesenheit von 3oxoC12-HSL zu einer erniedrigten Expression der Reifungsmarker CD80, CD86, CD83, CD40, HLA-DR, sowie der Migrationsmarker CD184 (CXCR4) und CD197 (CCR7). Die Coinkubation mit Zytokin-Cocktail und 3oxoC12-HSL ergab eine Herabregulierung der Maturationsmarker CD80, CD86 und HLA-DR. Auf unstimulierte DZ zeigte 3oxoC12-HSL keinen Effekt, das Oberflächenmarker-Expressionsprofil dieser Zellen glich dem unreifer DZ. 3oxoC12-HSL inhibierte auch die Sekretion der pro-inflammatorischer Zytokine IL-12, IFN-gamma, MIP-1alpha, TNF-alpha durch LPS- bzw. Zytokin-Cocktail-gereifte DZ. Insgesamt zeigen unsere Ergebnisse, dass 3oxoC12-HSL die Reifung von DZ unterdrückt und somit das Zustandekommen einer effektiven Immunantwort verhindert wird.

Dendritic cells (DCs) are bone marrow-derived professional antigen-presenting cells that act as master regulators of acquired and innate immune responses. Here, we review the available information on their role in human renal inflammation. In the 1980s and early 1990s, major histocompatibility complex class II antigen-(HLA-DR) positive DCs were first described in normal human kidneys and in the interstitium of kidneys from patients with glomerulonephritis. Several DC subtypes were subsequently distinguished based on their expression of CD1c/BDCA-1, CD141/BDCA-3 and CD209/DC-SIGN (in combination with HLA-DR). These cells were almost exclusively found in the tubulointerstitium, with increased numbers seen during glomerulonephritis. It appears that the human renal tubulointerstitium harbors different DC types which allow the collection of both exogenous as well as endogenous antigens. Plasmacytoid DCs have a plasma cell-like morphology and were commonly found within nodular tubulointerstitial infiltrates. Follicular DCs are rarely seen, but show a predominant localization in organized infiltrates. CD207/langerin is a marker for Langerhans cells. Langerin-positive cells have been found in association with the collecting ducts and urothelium. A functional characterization of these subtypes has been hampered by the difficulty of obtaining samples for analysis. However, these studies are clearly required to define the role of DCs and DC subsets in the pathophysiology of renal disease. Copyright (C) 2011 S. Karger AG, Basel

Die bullöse Dermatose Pemphigoid gestationis ist eine äußerst seltene schwangerschaftsassoziierte Autoimmunerkrankung, bei der Antikörper gegen Strukturen der Plazenta produziert werden, die mit dem BP180 der mütterlichen Haut kreuzreagieren und schließlich zur Blasenbildung führen. Ziel der vorliegenden Arbeit war die Assoziation der Erkrankung mit sämtlichen klassischen Humanen Leukozyten-Antigenen (HLA) zu überprüfen. Dafür wurden HLA-Kompletttypisierungen (HLA-A, -B, -C, -DRB1, -DQB1, -DPB1) bei 18 betroffenen Patientinnen und falls vorhanden von Partnern und Kindern der Betroffenen vorgenommen. Von Kindern und Partnern standen insgesamt 27 DNA-Proben zur Verfügung. Diverse Studien wiesen eine Assoziation der Erkrankung mit den HLA-Klasse II-Allelen HLA-DRB1*03 und *04 nach. Eine pathogenetische Rolle paternaler HLA-Moleküle bei der Erkrankung wird aufgrund der Häufung des HLA-DRB1*02-Phänotyps im Kollektiv der Partner vermutet. Die statistischen Berechnungen der Typisierungsergebnisse dieser Arbeit zeigten im Kollektiv der PG-Patientinnen eine signifikante Häufung der HLA-Allele B*08 (p=0,0019), B*18 (p=0,0455), DRB1*03 (p=0,001), DQB1*02 (p=0,0283) und DPB1*0101 (p=0,0011). In Übereinstimmung mit Daten aus der Literatur trat im PG-Patientinnen-Kollektiv der DRB1*03/*04-Phänotyp und im Kollektiv der Partner der HLA-DR02-Phänotyp signifikant gehäuft auf. Untersuchungen auf Aminosäureebene zeigten Gemeinsamkeiten der HLA-DRB1-Allele, die in dem Kollektiv der Mütter vertreten waren auf. Bei der Betrachtung der Genkonstellationen kompletter Stammbäume war ersichtlich, dass im Fall von 3 der 6 aufgeführten Stammbäume die Vererbung des väterlichen DRB1*02-Alleles mit der Krankheitsentwicklung in der entsprechenden Schwangerschaft zusammenfiel. Es wurden keine Hinweise gefunden, dass die Interaktion von HLA-C bzw. HLA-B mit KIR (killer cell immunglobulin-like receptors) für das Auftreten der Krankheit eine Rolle spielen. Die Ergebnisse legen einerseits nahe, dass aufgrund der Häufung von Allelen verschiedener HLA-Loci mehrere Gene eine ätiopathogenetische Rolle spielen, andererseits die Häufung mehrerer Gene verschiedener Loci auf einer Kopplung beruhen könnte. Desweiteren liefern die Untersuchungen der Aminosäurensequenzen Hinweise darauf, dass sich bestimmte Aminosäurensubstitutionen in der Bindungsgrube der HLA-Klasse II-Moleküle als protektiv 72 oder als erhöhtes Risiko auswirken könnten. Die Rolle des väterlichen HLA-DR bleibt weiterhin ungeklärt. Die Konstellation der HLA-DR-Vererbung innerhalb kompletter Familien suggeriert, dass die Vererbung des HLA-DRB1*02 an das Kind als zusätzlicher Risikofaktor zu bewerten ist, aber nicht als ursächlich für die Erkrankung betrachtet werden kann. Zur weiteren Klärung der paternalen Rolle wäre die Überprüfung der HLA-Klasse II-Expression in PG-Plazenten mit modernen molekularbiologischen Methoden von großem Interesse.

Thu, 20 May 2010 12:00:00 +0100 https://edoc.ub.uni-muenchen.de/11548/ https://edoc.ub.uni-muenchen.de/11548/1/Beeck_Stefan.pdf Beeck, Stefan

Im Mittelpunkt der Arbeit stehen die Auswirkungen des Prostanoids Prostaglandin E2 (PGE2) auf die Fähigkeit von Monozyten-abgeleiteten dendritischen Zellen (MoDC) zur Antigenpräsentation auf MHC-I- und MHC II Molekülen. Die Kreuzpräsentation von exogenem Antigen auf MHC I Molekülen ist eine entscheidende Fähigkeit der dendritischen Zellen (DC), die es ihnen ermöglicht zytotoxische T-Zellen zu aktivieren. PGE2 wird in Aktivierungsprotokollen als „Goldstandard“ in Kombination mit Zytokinen, wie TNF-α, IL-1β und IL 6, als Reifestimulus für DC in klinischen DC-basierten Tumorvakzinierungsstudien verwendet. Bisher wurde über den Effekt von PGE2 auf die Antigenpräsentation von Tumorantigen in Proteinform nicht berichtet. In dieser Arbeit wurde die Auswirkung von PGE2 und spezifischen EP Rezeptoragonisten auf verschiedene Funktionen von MoDC, die für die Antigenpräsentation auf MHC-I- und MHC-II-Molekülen relevant sind, untersucht. Zuerst wurde die Auswirkung von PGE2 und den EP Rezeptoragonisten auf die Ausreifung der MoDC untersucht. Hierbei wurde ersichtlich, dass MoDC nach Inkubation mit PGE2, vermittelt über EP2/4 Rezeptoren, die Aktivierungsmarker HLA A, HLA-B und HLA-C3, HLA-DR, CD83 und CD86 hochregulieren. Als nächstes wurde der Einfluss von PGE2 auf die Antigenaufnahmefähigkeit der MoDC untersucht. Hierbei wurde die Phagozytosefähigkeit anhand von Zelllysat und apoptotischen Tumorzellen und die Makropinozytosefähigkeit anhand von Dextran Partikeln analysiert. Es konnte gezeigt werden, dass weder PGE2 noch spezifische EP Rezeptoragonisten die Antigenaufnahme signifikant beeinflussten. Durch Präsentation von Antigen auf MHC-II-Molekülen sind DC in der Lage, tumorspezifische CD4+ T-Zellen zu aktivieren. Die MHC-II Präsentationsfähigkeit der MoDC unter dem Einfluss von PGE2 wurde mit NY-ESO-1157-170 Peptid, NY-ESO-1 Protein sowie mit NY ESO 1-transfiziertem CHO Zelllysat als Antigenquellen untersucht. PGE2 wurde zu den MoDC entweder 24 h vor (Präinkubation) oder zur gleichen Zeit wie das Antigen (Simultaninkubation) hinzugefügt. Die Versuche ergaben, dass die MHC-II-Antigenpräsentation der MoDC durch PGE2 nicht signifikant beeinflusst wird. Die Auswirkung von PGE2 auf die Kreuzpräsentationsfähigkeit der MoDC wurde mit einer Formulierung aus NY-ESO-1 Protein und dem ISCOMATRIX® adjuvant (NY ESO-1/IMX) und einem Immunkomplex bestehend aus NY-ESO-1 Protein und dem Antikörper anti NY ESO-1 mAk (Klon ES121; NY-ESO-1/IC) untersucht. Hierbei konnte gezeigt werden, dass sowohl die Kreuzpräsentation von NY-ESO-1/IMX als auch von NY ESO 1/IC durch PGE2 dosisabhängig signifikant gehemmt wurde. Dieser Effekt wird, ähnlich wie die Zell-Reifung, über die EP2/4-Rezeptoren vermittelt. Zusammenfassend erlauben die Ergebnisse den Schluss, dass PGE2 über einen EP2/4-Rezeptor-vermittelten Mechanismus die Kreuzpräsentationsfähigkeit der MoDC inhibiert, ohne jedoch die Antigenaufnahme oder die MHC-II Präsentation signifikant zu beeinflussen. Eine klinische Relevanz der Ergebnisse besteht bei der Verwendung von DC für therapeutische Tumorvakzinierungen zur Induktion einer gegen Malignome gerichteten Immunantwort. Die Benutzung von PGE2 als Reifestimulus für MoDC, die mit Protein-Antigenen gepulst werden, birgt das Risiko, die Induktion von tumorantigenspezifischen CD8+ T-Zellen negativ zu beeinflussen.

Die vorliegende Arbeit beschäftigt sich mit der funktionellen und molekularen Charakterisierung von humanen CD34- Zelllinien aus dem peripheren Blut (V54/1, V54/2) im Vergleich zu den aus dem Knochenmark etablierten Zelllinien (L87/4, L88/5). Die Klone V54/1 und V54/2 wurden aus dem peripheren Blut nach Stammzellmobilisierung und CD6 Depletion durch Zugabe eines Faktorengemisches aus IL-1b, IL-3, IL-6, IL-7, IL-8 und IL-11 erzeugt. L87/4 und L88/5 hingegen sind adhärente und wachstumsarretierte Stromazellen, die die Erhaltung und Differenzierung von hämatopoetischen Vorläuferzellen durch Mediatoren ermöglichen (Thalmeier et al. 2000). Das Ziel dieser Arbeit war die Untersuchung von Stammzelleigenschaften bei den Zelllinien L87/4, L88/5, V54/1 und V54/2. Dazu soll die Färbung mit den Farbstoffen Rhodamin 123 (Rh123) und Hoechst 33342 zeigen, ob Subpopulationen innerhalb der Klone mit unterschiedlichen Färbeeigenschaften, bestehen. Die biologische Bedeutung der beiden Farbstoffe liegt darin, dass Sie dazu geeignet sind frühe Stammzellen zu identifizieren. Als Substrat der P-Glykoproteinpumpe, die u.a. auf frühen Vorläuferzellen mit stark erhöhter Repopulationskapazität gefunden wird, werden diese Farbstoffe aus der Zelle gepumpt. Der Farbstoff-Efflux kommt durch die mdr-Gen-kodierte (multi-drug-resistance) und Kalzium-abhängige P-Glykoproteinpumpe zustande. Das P-Glykoprotein hat neben der Bedeutung in der Stammzellbiologie in der angewandten Medizin eine wichtige Funktion in der Resistenzentwicklung von Tumoren. Des weiteren wurden bei den Zelllinien stammzellrelevante Oberflächenantigene (CD10, CD34, CD14, CD105, SH3 und CD117) untersucht, um Unterschiede zwischen L87/4, L88/5 und den Klonen V54/1, V54/2 zu erkennen. Versuche zur Induktion der Differenzierung sollten Hinweise auf die Plastizität der Zelllinien geben. Experimente an den durch den Rh123-Efflux unterscheidbaren Subpopulationen der Zelllinie V54/2 dienen der Aufklärung von Unterschieden in Morphe, zellulären Transportfunktionen und Funktionseinheiten von Transkriptionsfaktor Netzwerken. Methodisch wurde für die Analyse der Epitope und der Färbungen mit Rh123 und Hoechst 33342 ein Durchflußzytometer verwendet. Die Analyse der Funktionseinheiten von Transkriptionsfaktor Netzwerken wurde mittels Reverse Transkriptase Polymerase Ketten Reaktion durchgeführt. Die Ergebnisse der Färbeexperimente zeigten, dass bei allen untersuchten Zelllinien durch eine unterschiedliche Anfärbbarkeit der Zellen mit dem Farbstoff Rh123 zwei Subpopulationen unterschieden werden können. Die jeweils größere Subpopulation der Zelllinien färbt sich mit Rh123 an und bleibt auch nach einer definierten Inkubationszeit, die den Rh123-Efflux ermöglichen soll, gefärbt. Sie wird Rh123high genannt. Die übrigen Zellen, die bei allen Zelllinien unter 10% der Gesamtpopulation betragen, sind in der Lage den Farbstoff aus der Zelle zu pumpen. Diese Subpopulation wird Rh123low genannt und ist mit Stammzelleigenschaften wie tausendfach erhöhter Repopulationsfähigkeit in NOD/SCID-Mäusen assoziiert. Es konnte also innerhalb der untersuchten monoklonalen Linien eine Rh123low Subpopulation identifiziert werden, die sich durch zahlreiche biologische Eigenschaften von der Gesamtpopulation unterscheidet. Da der Rh123 Efflux durch eine Kalzium-abhängige Pumpe zustande kommt, lässt sie sich durch den Kalziumantagonisten Verapamil hemmen. Eine Hemmung der Pumpe bewirkt, dass die Rh123low Zellen nicht mehr in der Lage sind Rh123 aus der Zelle zu pumpen, so dass sie nach einer definierten Inkubationszeit mit Rh123 gefärbt bleiben. Neben diesem funktionellen Beweis für die P-Glykoproteinpumpe konnte durch den strukturellen Nachweis der Pumpe mittels eines Antikörpers gegen P-Glykoprotein ein definitiver Beweis für das Vorhandensein der aktiven P-Glykoproteinpumpe bei der Rh123low Population erbracht werden. Mit dem anderen Farbstoff Hoechst 33342 können die jeweiligen Anteile der Zelllinien in den einzelnen Stadien des Zellzyklus nachgewiesen und zudem ein kleiner Anteil an Zellen bestimmt werden, der als „Side Population“ (SP-Zellen) definiert wird. Diesen SP-Zellen werden Eigenschaften von aktiven Stammzellen zugeschrieben. Hierbei besteht ein Unterschied zwischen den aus dem Knochenmark und den aus dem peripheren Blut etablierten Linien, da die Zellen aus dem peripheren Blut nicht nur ein anderes Zellzyklusmuster aufweisen, sondern auch einen höheren Anteil an SP-Zellen besitzen. Es wurden vergleichende Untersuchungen zwischen den Zelllinien und zwischen den Rh123high und Rh123low Subpopulationen innerhalb einer Zelllinie mit Antikörpern gegen die Epitope CD14, CD45, HLA-DR, CD10, CD117, CD105 und SH3 durchgeführt. Dabei waren CD14 und CD45 auf allen Zelllinien negativ, wobei alle Zelllinien eine positive Expression für den mesenchymalen Marker Endoglin (CD105) und für SH3 (CD73) zeigten. CD117 konnte nur auf den aus dem Knochenmark etablierten Zelllinien L87/4 und L88/5 nachgewiesen werden. CD34, ein charakteristischer Marker für hämatopoetische Vorläuferzellen, aber auch für Endothelzellen, konnte nur auf den Zellen der Rh123low Subpopulation nachgewiesen werden. Im Gegensatz dazu exprimieren die Rh123high Zellen kein CD34. Da es sich bei den Zelllinien um Klone handelt, ist der Unterschied in der Expression von CD34 zwischen der Rh123low und der Rh123high Population ein deutlicher Hinweis auf die Plastizität der Zelllinien und das Fließgleichgewicht zwischen Rh123low und Rh123high. Durch eine Zellsortierung der Zelllinie V54/2 wurde die Rh123low von der Rh123high Subpopulation getrennt, um sie dann bezüglich ihrer Morphologie, dem Wachstum in Methylzellulose und der Expression ausgewählter Funktionseinheiten von Transkriptionsfaktor Netzwerken zu untersuchen. Dabei erhärtete sich die Hypothese, dass es sich bei der Rh123low Subpopulation um aktivere Zellen mit einer gesteigerten Expression von erythroid/myeloischen und mesodermalen Eingaben (z.B. VEGF, BMP-4), Rezeptoren (z.B. tie-1), vernetzter Transkriptionsfaktoren (z.B. GATA, ETS) und letztendlich Ausgaben (z.B. PECAM) handelt. Diese fungieren in Netzwerken mit dem Ziel, stammzellrelevante Funktionen zu ermöglichen. Die Morphologie zeigte in den Zytozentrifugationspräparaten deutliche Unterschiede zwischen Zellen der Rh123low und der Rh123high Subpopulation. Die Rh123low Subpopulation besteht aus lymphoid-ähnlichen Zellen, was für Zellen mit Stammzellfunktion charakteristisch ist. Die Rh123high Subpopulation dagegen hat ein insgesamt größeres Zellvolumen und einen gebuchteten Kern mit perinukleärer Aufhellung. Untersuchungen des klonalen Wachstums in der Methylzellulose ergaben bei keiner der Subpopulationen eine wesentliche Koloniebildung. Durch die Inkubation der Zelllinie V54/2 mit dem Neurotropen Wachstumsfaktor (NGF) konnte eine morphologische Änderung in Richtung einer neuronalen/glialen Differenzierung nach 8-12 Stunden induziert werden. Der immunhistochemische Nachweis von Glial Fibrillary Acidic Protein (GFAP) bestätigte die mesenchymale Potenz zumindest in Richtung einer glialen Differenzierung. Das unterschiedliche Expressionsmuster ausgewählter, für die Differenzierung notwendiger Zusammenspieler innerhalb von Transkriptionsfaktor Netzwerken innerhalb der Rh123high und der Rh123low Population bei V54/2 war ein weiterer Hinweis, dass es sich bei der Rh123low Subpopulation um aktive Vorläuferzellen mit möglicher Stammzellpotenz handelt. In der Rh123low Subpopulation wurde im Gegensatz zur Rh123high Population eine Expression von BMP4, GATA1, GATA3 nachgewiesen, die essentiell für die Hämatopoese und für eine mesenchymale Differenzierung ist. Die Faktoren für GATA2, GATA3, beta globin, Elf-1 und PECAM1 wurden in einem stärkeren Maß in der Rh123low als in der Rh123high Population exprimiert. BMP-Rez., Myb, sowie die Endothel-assoziierten Faktoren Tie-1 und VEGF waren in beiden Subpopulationen gleich stark vorhanden. Bei den wenigen Funktionseinheiten der größeren und Rh123high Population handelt es sich vor allem um angiogenetische Faktoren, was auf eine limitierte Differenzierungseigenschaft der Rh123high Subpopulation und die enge Beziehung zwischen Blut- und Endothelzellen („Hämangioblast“) hinweist. Ein Nachweis für die Plastizität der Stammzellen innerhalb der von uns etablierten Zelllinien wurde dadurch erbracht, dass die zellsortierten Subpopulationen Rh123low und Rh123high nach dem Sortierexperiment getrennt rekultiviert wurden, wobei das Wachstum der Rh123low Subpopulation deutlich langsamer war als das der Rh123high Subpopulation. Nach zwei Wochen wurden die zellsortierten Subpopulationen erneut einer Rh123 Färbung unterzogen, wobei sich wiederum das ursprüngliche Verhältnis zwischen den Rh123low und Rh123high Subpopulationen einstellte. So kann man aus der Transdifferenzierung der Zelllinien von Rh123low in Rh123high und umgekehrt die Plastizität der hier untersuchten adulten Stammzelllinien ableiten. Die Ergebnisse sollen zum grundlegenden Verständnis der Biologie adulter (nicht embryonaler) Stammzellen beitragen und damit die Möglichkeit schaffen, adulte Stammzellen bzw. deren Subpopulationen gezielt für einen reparativen Gewebe- und Organersatz zu verwenden. Dabei liefern sie die Basis für weitergehende Untersuchungen zum besseren Verständnis der physiologischen und regenerativen Vorgänge, z.B. auch bei Alterung oder bei gesteigerter Funktion. Darüber hinaus kann aufgrund der vorliegenden Ergebnisse durch weitere Untersuchungen möglicherweise besser verstanden werden, ob es gelingen kann das Potential adulter Stammzellen zur therapeutischen Gewebereparation, z.B. zur Verhinderung oder Verringerung einer Narbenbildung, zu nutzen.

Schizophrenia has often been observed to be associated with alterations of the cellular immune system. In this study we monitored numerous immune parameters in the course of antipsychotic treatment. 40 patients diagnosed with an acute exacerbation of schizophrenia were tested before and during treatment with antipsychotics. The percentual distribution of lymphocyte subgroups (CD3, CD4, CD8, CD19, CD5, CD16/56, CD25, HLA-DR, gd, CD11a, CD11b and CD49d) was measured by FACS analysis. 20 healthy volunteers served as controls. In the acute state of psychosis a significant elevation of the B-lymphocyte fraction was observed, while the percentage of T-lymphocytes was decreased. These values levelled to those of the control group in the course of treatment. Natural killer cells, integrine expressing cells and CD5+ B-cells showed alterations in numbers after several months of treatment. The latter may be a side effect of antipsychotic treatment. The findings in the acute state of schizophrenia could be seen as a nonspecific stress reaction or an effect of the altered metabolism of neurotransmitters. On the other hand the observed changes in the immune system could be due to infectious diseases or autoimmune processes provoking the symptoms of schizophrenia.

Bei schizophrenen Patienten sind immunologische Veränderungen nachweisbar, die auf eine Aktivierung des Immunsystems hinweisen und die seit langem diskutierte Hypothese stützen, dass das Immunsystem in der Pathogenese der Schizophrenie involviert ist. Es gibt Anhaltspunkte, die die Annahme eines inflammatorischen - sei es infektiös oder autoimmunologisch getriggerten - Prozesses im Gehirn schizophrener Patienten stützen, der zu einer erhöhten Permeabilität der Blut-Hirnschranke führt und für die psychopathologische Symptomatik verantwortlich gemacht wird. In dieser Arbeit wurden mittels Dreifarben-Durchflusszytometrie (FACS-Analyse) Lymphozyten-Subpopulationen von 31 schizophrenen Patienten bestimmt und sowohl mit Befunden gesunder Kontrollpersonen, als auch desselben Patienten vor und nach Neuroleptikabehandlung verglichen. Gemessen wurde die Expression einer Auswahl von Oberflächenantigenen, die den Aktivierungszustand des Immunsystems anzeigen bzw. mit autoimmunologischen Prozessen in Verbindung gebracht werden. Dadurch war es möglich allgemeine immunologische Veränderungen des peripheren Immunsystems schizophrener Patienten zu erfassen und diese auch im Verlauf der Therapie mit Neuroleptika, sowie in Korrelation mit psychopathologischem Befund und Prognose zu betrachten. Die in dieser Arbeit gezeigten Ergebnisse stützen die Hypothese, dass bei schizophrenen Patienten eine Aktivierung des Immunsystems vorliegt. So fand sich bei Schizophrenen nach Behandlung im Vergleich zu Kontrollen eine Erhöhung der CD4-positiven T-Zellen – ein Befund, der auch bei Autoimmunerkrankungen gesehen wird und dort für die chronische Stimulation von B-Lymphozyten verantwortlich gemacht wird. Zusätzlich wurden Patientengruppen anhand des psychopathologischen Verlaufes und des Therapieerfolgs gebildet. Ein Zusammenhang bestand zwischen dem Ansprechen auf Therapie und CD8-positiven T-Lymphozyten: Die Patientengruppe mit niedrigerem Anteil von CD8+- Zellen, also weniger Suppressorzellen, zeigte ein schlechteres Ansprechen. Ein weiteres Aktivierungszeichen des zellulären Immunsystems ist die Expression des CD45RO-Moleküls anstelle von CD45RA auf CD4-positiven Lymphozyten. Auch hier konnte bei schizophrenen Patienten ein erhöhter Anteil von CD4+/CD45RO+- (Gedächtnis-) Zellen, insbesondere nach Behandlung, beobachtet werden, wobei wiederum Patienten mit schlechterem Ansprechen auf Therapie stärkere Veränderungen zeigen. Unterstrichen wird dieser Befund durch die parallele Zunahme der Expression des Adhäsionsmoleküls CD58 und CD60 bei Schizophrenen unter Behandlung, sowie zusätzlich bei schlechter Therapieantwort. Eine Expression dieser Moleküle weist auf die Fähigkeit zur transepithelialen Migration der Zellen, auf eine weitere Immunaktivierung und Stimulation von B-Zellen hin. Ebenfalls ähnlich wie bei Autoimmunerkrankungen wurde in der hier untersuchten Patientengruppe eine erhöhte Expression von HLA-DR gefunden. Dies, wie auch eine geringe Zahl von CD4+/45RA+-Lymphozyten, die als Suppressor-Inducer-Zellen bekannt sind, kann als fehlende Suppression von Autoimmunreaktionen interpretiert werden. Darüberhinaus scheint ein schlechtes Outcome der Patienten mit insgesamt erhöhter immunologischer Aktivität assoziiert zu sein. Die Ergebnisse im Bezug auf CD5+/19+- Zellen und NK-Zellen runden das Bild einer Modulation immunologischer Mechanismen im Verlauf einer schizophrenen Störung und durch neuroleptische Therapie ab. In der Literatur beschriebene Störungen der Blut-Hirnschranke und Reaktionen des hirneigenen Immunsystems weisen auf eine Interaktion von peripherem und zentralem Immunsystem im Rahmen der Erkrankung hin. In der akuten Phase der Schizophrenie scheint dabei die Aktivierung des Th2-Systems im Vordergrund zu stehen. Im Verlauf der neuroleptischen Behandlung findet dann ein Wechsel zum Th1-System statt. Veränderungen im Zytokinsystem sind von besonderem Interesse, da diesen Botenstoffen eine Vermittlerrolle zwischen Immunsystem und Nervensystem zukommt und so ein Einfluss auf neurometabolische Vorgänge erklärbar ist. Die in dieser Arbeit beobachteten Veränderungen unter Neuroleptikatherapie sprechen für einen immunmodulatorischen Wirkmechanismus dieser Substanzen. Für die Substanz Clozapin sind beispielsweise derartige Effekte bekannt. Bei der Schizophrenie handelt es sich um eine heterogene Krankheitsentität, für die es zahlreiche Erklärungsmodelle gibt. Die hier erhobenen Befunde stehen in Einklang mit Ergebnissen anderer Studien und können diese zum Teil bestätigen und ergänzen. Allerdings finden sich in der Literatur diskrepante und umstrittene Befunde bezüglich der zellulären und humoralen Veränderungen des Immunsystems, die die Komplexität des Krankheitsbildes deutlich machen und vielleicht als Ausdruck ätiologischer Unterschiede angesehen werden können.

Dendritic cells (DC) constitute a heterogeneous leukocyte population having in common a unique capacity to induce primary T cell responses and are therefore most attractive candidates for immunomodulatory strategies. Two populations of blood DC (CD11c+ CD123(dim) and CD11c- CD123(high)) have been defined so far. However, their direct isolation for experimental purposes is hampered by their low frequency and by the lack of selective markers allowing large scale purification from blood. Here we describe the monoclonal antibody (mAb) M-DC8, which was generated by immunizing mice with highly enriched blood DC. This mAb specifically reacts with 0.2-1% of blood leukocytes and enables their direct isolation by a one-step immunomagnetic procedure from fresh mononuclear cells. These cells can be differentiated from T cells, B cells, NK cells and monocytes using lineage-specific antibodies. M-DC8+ cells express HLA class It molecules, CD33 and low levers of the costimulatory molecules CD86 and CD40. Upon in vitro culture M-DC8+ cells spontaneously mature into cells with the phenotype of highly stimulatory cells as documented by the upregulation of HLA-DR, CD86 and CD40; in parallel CD80 expression is induced. M-DC8+ cells display an outstanding capacity to present antigen. In particular, they proved to be excellent stimulators of autologous mixed leukocyte reaction and to activate T cells against primary antigens such as keyhole limpet hemocyanin. Furthermore, they induce differentiation of purified allogeneic cytotoxic T cells into alloantigen-specific cytotoxic effector cells. While the phenotypical analysis reveals similarities with the two known blood DC populations, the characteristic expression of Fc gamma RIII (CD16) and the M-DC8 antigen clearly defines them as a novel population of blood DC. The mAb M-DC8 might thus be a valuable tool to determine circulating DC for diagnostic purposes and to isolate these cells for studies of antigen-specific T cell priming. Copyright (C) 2000 S. Karger AG, Basel.

A prospective study of a normal childhood population identified 44 islet cell antibody positive individuals. These subjects were typed for HLA DR and DQ alleles and investigated for the presence of antibodies to the Mr 64,000 (64K) islet cell antigen, complement-fixing islet cell antibodies and radiobinding insulin autoantibodies to determine their potency in detecting subjects with impaired Beta-cell function. At initial testing 64K antibodies were found in six of 44 islet cell antibody positive subjects (13.6%). The same sera were also positive for complement-fixing islet cell antibodies and five of them had insulin autoantibodies. During the follow-up at 18 months, islet cell antibodies remained detectable in 50% of the subjects studied. In all six cases who were originally positive, 64K antibodies were persistently detectable, whereas complement-fixing islet cell antibodies became negative in two of six and insulin autoantibodies in one of five individuals. HLA DR4 (p < 0.005) and absence of asparic acid (Asp) at position 57 of the HLA DQ chain (p < 0.05) were significantly increased in subjects with 64K antibodies compared with control subjects. Of 40 individuals tested in the intravenous glucose tolerance test, three had a first phase insulin response below the first percentile of normal control subjects. Two children developed Type 1 (insulin-dependent) diabetes mellitus after 18 and 26 months, respectively. Each of these subjects was non-Asp homozygous and had persistent islet cell and 64K antibodies. We conclude that 64K antibodies, complement-fixing islet cell antibodies and insulin autoantibodies represent sensitive serological markers in assessing high risk for a progression to Type 1 diabetes in islet cell antibody positive non-diabetic individuals.

Islet cell antibodies (ICAs) were determined in a large cohort of white nondiabetic schoolchildren (n = 4287) from a homogenous population in southern Germany. The prevalence of ICA levels greater than or equal to 5 Juvenile Diabetes Foundation (JDF) U was 1.05% (95% confidence interval 0.8-1.4%). Analysis of HLA-DR beta and -DQ beta alleles revealed that the specificities found to be increased in insulin-dependent (type I) diabetic subjects with the same ethnic background were also associated with ICA positivity in the nondiabetic schoolchildren. HLA-DR3 (P less than 0.01) and -DR4 (P less than 0.01) phenotypes and absence of Asp residue (P less than 0.01) at codon 57 of the HLA-DQ beta-chain were significantly increased in ICA+ compared with control subjects. High levels of ICAs, which were categorized as either greater than or equal to 17 or greater than or equal to 30 JDF U, were found to be associated with amino acids other than Asp at position 57 of the HLA-DQ beta-chain. No association of ICA level was found for HLA-DR phenotypes.

Mon, 1 Jan 1990 12:00:00 +0100 https://epub.ub.uni-muenchen.de/7965/1/7965.pdf Goebel, Frank-Detlef; Nerl, C.; Schreiber, M. A.; Heinrich, B.; Matuschke, A.; Bogner, Johannes R.

Induction of clonal anergy in T-helper (Th) cells may have a role in regulating immune responses. A model system for studying Th cell tolerization at the clonal level in vitro could be useful for investigating the mechanisms involved. Accordingly, alloreactive helper cells were maintained in culture with interleukin 2 (IL 2) by intermittent stimulation with specific antigen. Regardless of the frequency of antigen stimulation, clones of age less than ca. 35 population doublings (PD) were found to undergo antigen-specific autocrine clonal expansion in the absence of exogenous IL 2. Such young clones (designated as phase I) could therefore not be "tolerized" by frequent exposure to antigen. In contrast, most clones of age greater than ca. 35 PD could be tolerized by frequent exposure to antigen (designated as phase II clones). Their autocrine proliferation was then blocked, although they still recognized antigen specifically as shown by their retained ability to secrete interferon-gamma (IFN-gamma) and granulocyte-macrophage colony stimulating factor (GM-CSF). The mechanism of response failure involved both an inability to upregulate IL 2 receptors in the absence of exogenous IL 2, as well as an inability to secrete IL 2. These defects were not overcome by stimulation with mitogens or calcium ionophore and phorbol esther in place of alloantigen. T-cell receptor, alpha, beta, and gamma-chain gene rearrangements remained identical in phase I and phase II clones. Tolerization of phase II clones could be avoided by increasing the period between antigen exposures. Despite this, whether or not phase II cells were capable of autocrine proliferation, they were found to have acquired the novel function of inducing suppressive activity in fresh lymphocytes. Suppressor-induction was blocked by the broadly reactive MHC class II-specific monoclonal antibody (moAb) TU39, but not by moAb preferentially reacting only with HLA-DR, DQ, or DP. Sequential immunoprecipitation on T-cell clones showed the presence of a putative non-DR, DQ, DP, TU39+ molecule on phase II clones. However, this molecule was also found on phase I clones. The nature of the TU39-blockable suppressor-inducing determinant present on phase II but not on (most) phase I clones thus remains to be clarified. In addition to suppressor-induction activity, phase II clones also acquired lytic potential as measured in a lectin approximation system. Cytotoxic (CTX) potential was also not influenced by the frequency of antigenic stimulation and could be viewed as a constitutive modulation of clonal function

A set of T cell clones (TCC) isolated from HLA-DR-, Dw-, DQ-matched allogeneic MLCs was found to proliferate autonomously when stimulated with cells carrying a wide range of class I or II specificities. This apparently unrestricted proliferation was relatively weak, and only low levels of IL-2 were present in the supernatants of stimulated cells. Autologous as well as allogeneic PBMC and B lymphoblastoid cell lines (B-LCL) were capable of stimulating such clones, which were also restimulated by suppressive, but not by helper, TCC. Moreover, such clones displayed the unusual property of autostimulation. mAb inhibition experiments suggested that class II- or class II-restricted antigens were involved in stimulation. Thus, certain "broad" mAbs (TU39, SG520) reacting with multiple locus products inhibited activation of these reagents, but none of those reacting more specifically with DR (TU34, TU37, L243, Q2/70, SG157), DQ (TU22, SPV- L3, Leu 10), or DP (B7/21), or mixtures of these mAbs, were able to do so. Evidence from sequential immunoprecipitation experiments suggested that mAb TU39 bound class II-like molecules other than DR, DQ, and DP on TCC and B-LCL, and it is therefore proposed that such putative novel class II-like molecules may carry the stimulating determinants for these autoreactive clones. DY-reactive clones lacked helper activity for B cells but mediated potent suppressive activity on T cell proliferative responses that was not restricted by the HLA type of the responding cells. Suppressive activity was induced in normal PBMC by such clones, as well as by independent suppressive clones, which was also inhibited only by mAb TU39. These findings lead to the proposal that DY-reactive autostimulatory cells may constitute a self- maintaining suppressive circuit, the level of activity of which would be regulated primarily by the availability of IL-2 in the microenvironment