Podcasts about cd11b

BUFFALO, NY - March 31, 2025 – A new #research paper was #published in Oncotarget, Volume 16, on March 21, 2025, titled “FGR Src family kinase causes signaling and phenotypic shift mimicking retinoic acid-induced differentiation of leukemic cells." A research team led by first author Noor Kazim and corresponding author Andrew Yen from Cornell University discovered that the FGR protein—traditionally considered a cancer-promoting molecule—can instead trigger leukemia cells to mature. This effect mirrors the response usually induced by retinoic acid (RA); a compound derived from vitamin A that is widely used in cancer therapy. Their finding presents a potential new path for therapies targeting acute myeloid leukemia (AML) and related cancers. Acute myeloid leukemia is often treated using RA-based therapies that force immature white blood cells to mature, slowing their rapid growth. Retinoic acid works through complex signaling and gene regulation involving a group of proteins that orchestrate this transformation. In this study, the team used HL-60 cells, a model for human leukemia, and engineered them to express FGR. Surprisingly, the presence of FGR alone was enough to make these cells mature in a way almost identical to what happens with RA treatment. They began producing well-known markers of maturation such as CD38 and CD11b, generated reactive oxygen species (ROS), and expressed the inhibitor of the cell cycle, p27, all signs that the cells had shifted from a cancer-like, fast-dividing state to a more specialized, mature form. Further analysis revealed that FGR activated a group of proteins known as the "signalsome," which helps trigger the changes needed for cells to differentiate. This same group is typically activated by RA. “Notably, FGR induces the expression of genes targeted by RAR/RXR, such as cd38 and blr1, even without RA." To test its potential use in treatment-resistant leukemias, the researchers introduced FGR into RA-resistant HL-60 cells. In these, FGR did not cause the same maturation process, which suggests that there are other problems with cell signaling that stop both the RA and FGR pathways. This result highlights the complexity of resistance mechanisms and the need for additional research. These findings challenge the traditional view of FGR as strictly a cancer-driving protein. Instead, in this specific context, it appears to initiate anti-cancer behavior. That a single protein can reproduce the effects of a complex therapeutic compound like RA is both surprising and promising. If future research confirms this study's results in more advanced models, FGR could become a new tool for developing therapies for AML and potentially other blood cancers. DOI - https://doi.org/10.18632/oncotarget.28705 Correspondence to - Andrew Yen - ay13@cornell.edu Video short - https://www.youtube.com/watch?v=v2fjeFFoUPQ Subscribe for free publication alerts from Oncotarget - https://www.oncotarget.com/subscribe/ 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. 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@IMPACTJOURNALS.COM

Superimposed on an impressive body of work on the blood-brain-barrier and immune system, Prof Akassoglou and her collaborators just published an elegant study in Nature that centered on the direct binding os the SARS-CoV-2 spike protein to fibrin with marked downstream pro-inflammatory effects. The findings and potential treatments have implications beyond Covid, Long Covid to other neurologic diseases.Full videos of all Ground Truths podcasts can be seen on YouTube here. The audios are also available on Apple and Spotify.Transcript with links to audio and to relevant papers, graphicsEric Topol (00:07):Well, hello this is Eric Topol with Ground Truths, and with me today is Katerina Akassoglou. She is at the Gladstone Institute and she is a remarkable neuroimmunologist who has been doing extraordinary work for three decades to unravel the interactions between the brain, blood vessels and the role of inflammation. So Katerina, there's a lot to discuss, so welcome.Katerina Akassoglou (00:40):Thank you. Thank you so much. It's a great pleasure to join.By Way of BackgroundEric Topol (00:43):It's really interesting going back in your career. First of all, we're thankful that you immigrated here from Greece, and you have become one of the leading scientists in this discipline of important discipline of neuroimmunology, which is not just about Covid that we're going to talk about, but Alzheimer's and neurodegenerative diseases. This is a really big hot area and you're definitely one of the leaders. And what I was impressed is that all these years that you've been working on the integrity of the blood-brain barrier, the importance of fibrinogen and fibrin, and then comes along the Covid story. So maybe what we can do is start with that, which is you've made your mark in understanding this whole interaction between what can get into the brain, through the blood-brain barrier and incite inflammation. So this has been something that you've really taken to the extreme knowledge base. So maybe we can start with your work there before we get into the important seminal Nature paper that you recently published.Katerina Akassoglou (01:57):Yes, of course. So since very early on, I was still a graduate student when we made the first discovery and at the time was like mid-90s, so it was really ahead of its time. That dysregulation of cytokine expression in the brain of mice was sufficient to induce the whole cascade of events, triggering neurodegeneration, demyelination in pathological alterations, very reminiscent of multiple sclerosis pathology. And it was really hard to publish that study at the time because it was not yet accepted that this regulation of the immune system modeling the brain can be linked to neurodegeneration. So that was 1995 when we made that discovery, and I became really interested, what are the pathogenic triggers that actually polarized the immune cells in the brain? So with this, of course, this transgenic animal was expressing TNF, it was an artificially made animal that we made, but naturally what were the triggers that would polarize the innate immune cells? So I looked really early on in this mice and what I found was that the very first event was leaks of blood-brain barrier. It was opening of the blood-brain barrier in this mouse before inflammation, before demyelination, before neuronal loss. And this is really what shaped the question that, is it possible that these blood leaks that happened very early in the pathology, could this be the instigators of pathogenic inflammation in the brain?Eric Topol (03:34):Yeah. So in a way, you got at this question because of the chicken-and-egg and what happens first, and you got to the temporal saying, which happened first as you said, the leak before you could see evidence of inflammation and being able to study this of course in the experimental model, which you couldn't really do in people. And what I love about the description of your career, which has been quite extraordinary contributions is connecting the dots between the blood, the inflammatory response and the brain. Perhaps no one has done that like you have. And before we get into the recent paper, a lot of people are not aware that a year ago, a group in the UK known as PHOSP-COVID, they published a really important paper in Nature Medicine of over 1,800 people who were hospitalized with Covid and they found that fibrinogen was the best marker for cognitive deficits at 6 and 12 months (Figure below)(04:40):So that's just one of many papers, but it's a particularly well done study that already before you got into this work that recently published had emphasized fibrinogen. And by the way, again, having spent a lot of years in clots in the arteries, for me, we have to just get it down to fibrinogen plus thrombin gets you to fibrin. Okay, so fibrin is a major player here when fibrinogen is cleaved. So here we have the basis that you established, which is the fibrinogen leakage into the brain, activating inflammation, activating microglia, which like the macrophages of the brain and inciting the whole process. And before we close, I want to not just talk about Covid, but Alzheimer's too. But now let's get into the study that you did, [Fibrin drives thromboinflammation and neuropathology in COVID-19] which is striking, I mean really striking. And can you kind of take us through, because you not only demonstrated the importance of fibrin in inciting neuroinflammation in this model, but also how you could reverse it or prevent it. So this, and you looked at it in many different ways, this was a systematic approach. Maybe you can take us through how you were able to make such compelling evidence.The Multimodal EvidenceKaterina Akassoglou (06:09):Yes, thank you. First of all, thank you for bringing up the human relevance because this was also our inspiration for the work that we did in the Covid study. So as you mentioned in Covid patients, fibrinogen unbiased mass spec analysis was identified as the predictive biomarker for cognitive impairment in Long Covid patients. And this was in addition to also neuropathology data about the abundance of fibrin deposition in the brain. And these were studies that were done by NIH that have found deposition of fibrin in the brain and the reports for the abnormal and puzzling coagulation in Covid that is not setting other infections and also in many cases not always relating with the severity of symptoms. So even mild cases of Covid also had increased coagulation. I was really intrigued by this human, all this evidence in human data, and I thought that maybe the way that we're thinking about this, that it's systemic inflammation that drives the clotting.(07:24):Maybe there's another aspect to this. Maybe there is a direct effect of the virus with the coagulation cascade, and in this way maybe this can be an instigator of inflammation. So this was the original idea to be able to reconcile this data from the clinic about why do we have this prevalence of coagulopathy in Covid. And of course, the second question is, could this also be a driver of the disease? And of course, we're in a unique position because we have been studying this pathway now for over 20 years to have all the toolbox, the genetic toolbox, the pharmacologic toolbox to be able to actually really address these questions with genetic loss of function studies, with a blood innate immunity multiomics pipeline that we have set up in the lab. And of course, with preclinical pharmacology in our ABSL3 facility. So we had the infrastructure in place and the source in place to actually really dissect this question with both genetic tools as well as also technology platforms.Eric Topol (08:29):And you had in vivo imaging, you're the director of in vivo imaging for Gladstone and UCSF. So you do have the tools to do this.Katerina Akassoglou (08:38):Yes. The imaging that you mentioned is really important because this is, we employed that very early in our studies over now 15 years ago. And the reason was sometimes from snapshots of histopathology, you cannot really understand the sequence of events. So by being able to image these processes, both neuronal activity, microglia activation, infiltration of peripheral cells in the brain, this is how we could see the steps that what happens early on and to be able to answer these chicken-and-egg questions that you mentioned. So these were very, they're very important experiments, especially at the beginning because they were hypothesis driving and we were able to ask the right questions to drive our research program.Eric Topol (09:26):Now was the binding of the spike protein to one key site in fibrinogen, was that known before? [See outstanding Figure below from Trends in Immunology]Katerina Akassoglou (09:36):No, this was not known. So there was evidence that there are abnormal clots in Covid, but it was not known whether the spike protein would directly bind to protein to the coagulation cascade. So one of the key discoveries in our study was to use peptide array mapping and be able to identify not only the binding, but exactly the domains on fibrin that spike binds too. And what we found was two key domains, one the inflammatory domain and the other the plasmin binding site, which is important for fibrin degradation. So this suggested a potential dual deleterious role for this interaction, both by maybe affecting inflammation, but also delaying fibrinolysis, which is the degradation of this toxic protein from the brain. And indeed, we found that this interaction was responsible for all these two aspects, including decreased degradation, more inflammation, but also at the same time increased, increased coagulation. So it was a really pathogenic interaction.Eric Topol (10:47):Yeah, actually it's pretty striking. You have these two sites, the plasmin cleavage site of fibrinogen, which as you say, we knew there was a problem with clots. We knew that, but we didn't know exactly the spike protein how exactly it was implicated, particularly with fibrinogen. And then this other site, the CD11b-C18, now that's fancy for surface receptors of macrophages. And basically, this is critical because it's this microglia activation in the brain, and I know you saw it in the lungs as well through this other site that spike protein activated. So you had a twofer here of things that you discovered that the SARS-CoV-2 spike protein was capable of doing. This was a really big revelation. And then you also looked at mice that were genetically manipulated. So maybe you can, because before we get to your antibody monoclonal, the ways that you proved this were, I mean, one thing after another is really systematic. So maybe you can teach us about that.Thanks for reading Ground Truths! This post is public so feel free to share it.Establishing CausalityKaterina Akassoglou (12:08):Yeah, sure. So the first was about chemistry experiment. So this of course, we had to get to the next step to see is there any causality for this pathway. So we employed genetic loss of function studies and we had knockout mice, either fibrinogen knockout mice, this mice have all blood proteins except fibrinogen, and they have a delay in coagulation so they don't clot properly. But we also had a mutant mouse, which is a fibrinogen NK mouse. And this was a mutation only within this inflammatory domain that you mentioned, inflammatory domain that binds to C11b-C18. Other names for this is of course complement receptor 3, Mac-1 (αMβ2). It's the same, many names for this receptor, that as you mentioned, is expressed not only in microglial in the brain, but also peripheral immune cells including macrophages as well as also neutrophils which are CD11b expressing.(13:12):So we now have genetic models to be able to look at both complete depletion of fibrinogen, but also a very specific mutation and very selective mutation that only blocks the inflammatory properties without affecting the properties of fibrin in hemostasis. And these mice were made many years ago by a very close collaborator, Jay Degen at the University of Cincinnati. So what we found is that when we block either the inflammatory domain or we completely deplete fibrinogen, there was this profound protection after infection in internasal infection with the virus in lung inflammation. And this was both suppression of oxidative stress and this pathogenic inflammation in the lung, but also decreasing fibrosis, which has been associated with also Long Covid. And the surprise came from the transcriptomic data. So when we did transcriptomic analysis in this mice in the lungs, we found perhaps the expected decrease in the immune signatures in macrophages. This was in line with our previous work in, as you mentioned, Alzheimer's models, multiple sclerosis models. But what also was really surprising is there was that genes that are associated with activation of NK cells were upregulated. And of course this was the first time we had infected these mice, previously we had not done an infection before. So I think that maybe because of this region we had not seen before in our data this immunomodulatory role of fibrin that not only surprises the macrophage response, but also increases these NK cells that are important for viral clearance.Eric Topol (15:00):So again, the finding another important unique finding is the natural killer (NK) cells and effect there from the activation of this, as you said, the inflammation site or the CD11b-C18 that we've been talking about. So now another layer of this, a dimension of your Nature paper was that you tested an antibody that you already had developed so-called 5B8. A monoclonal that specifically binds to the domain of the one we're talking about this inflammation domain of fibrinogen. So can you tell us about what that showed?Katerina Akassoglou (15:45):Yes, so we tested this antibody in different models of Covid, which were both models with neuroinvasion and models without neuroinvasion. So we used both transgenic mice for hACE2, the human ACE2 infected with Delta, but we also use mouse adapted viruses like Beta that is just in the wild type mice with no transgenic being involved that these are without neuroinvasion. And we wanted to see if the antibody had any potential protective effects. And what we found is that the antibody protected from inflammation in the lung. So the data looked so similar with a genetic mutation of this pathway, protection from inflammation, decreased fibrosis, increased viral clearance, so decreased spike and viral proteins in the lungs. But we also found a protection in the brain. So the brains of this mice, including both the models we used with neuroinvasion and without, they both have had microglia activation in the brain. And we also found neuronal loss in the Delta infected mice and the antibody protected from both neuroinflammation but also improved neuronal survival in the mice. Showing that there can be this despite regardless of which model we used, there was this protective effect suggesting that by blocking fibrin, either the periphery or in the brain, this could be protected for these models.Eric Topol (17:28):Yeah, so I mean this is fascinating because until now, until this report of yours and your colleagues at Gladstone, there was knowledge that there would be neuroinflammation from Covid, both in patients from various biomarkers and imaging as well as in experimental model. But what this did was take it to the fibrin story, and I guess that's one of the questions you nailed that how important fibrin is, but that doesn't necessarily rule out other triggers of neuroinflammation, right?Katerina Akassoglou (18:04):Oh, absolutely not. So I think that this is one of the mechanisms that can be very important, especially in some patients. But we know that there are additional of course mechanisms of neuroinflammation including auto-antibody responses, as well as also endotheliopathy that are persistent endotheliopathy, this can be interacting also with each other. So I think that it's important for future research that we understand how do these mechanisms feed into each other? Are there a positive feedback loops between autoimmune mechanisms and coagulopathy and endothelial dysfunction with inflammation? But I think most importantly, I think that if we're thinking of this in the context of patients, can we identify patients with mechanism that might be more prevalent in specific cases of Long Covid and tailor our potential future clinical trials towards the needs of Long Covid patients?Towards TreatmentEric Topol (19:06):Absolutely. I did interview some months back on Grounds Truths, Michelle Monje at Stanford, who I'm sure and interact with, and she's also works not so much on the fibrin side, but on neuroinflammation and the likeness between this condition in people and chemo brain because of the inflammation that's seen there. So we've talked about the multiple triggers that could contribute to brain inflammation, which I think most people would say in Long Covid this is one of the most, besides obviously the lack of energy, the profound fatigue and disability, but the cognitive function hit, not just brain fog is often profound. And we've just seen some reports about that, and particularly in hospitalized patients, how bad that can be. So that gets us to a potential treatment. Now, one of the things that's out there dangling, there's many things that people have talked about in terms of why can't we have a treatment for Long Covid?(20:13):And now of course this fibrin pathway, if you will, lends itself to many possibilities, whether it's anticoagulants or fibrinolytics like a tPA or things like nattokinase, which is a Japanese food enzyme that you could get at the nutrition centers or whatever. What are your thoughts? Because we don't have any good studies. There are all these little, tiny studies and they don't provide much conclusion, and you have an antibody that could potentially be effective. As I understand it, you set up a company some years ago, Therini Bio and used to be called MedaRed. You're the first woman scientist at Gladstone to develop a spin out company, which is another point of congratulations on that. But could the antibody be tested in patients or what do you think about these other possibilities?Katerina Akassoglou (21:15):Yes, yes. These are great questions. So first of all, the different approaches that you mentioned have very different mechanism of action. So degrading fibrin, the degradation products of fibrin also can have deleterious effects. The dimer, for example, can be very pro-inflammatory. So at the same time, blocking coagulation can also have a diverse effects because this can lead to excessive hemorrhage. So the approach that we took was to selectively block the inflammatory properties of fibrin without affecting beneficial effects of the molecule in normal hemostasis. So the challenge when I made the antibody was to be able to dissect these two functions of fibrin. It's our most important clotting factor, but at the same time, a molecule with profound pro-inflammatory capacity. So the observation that these two domains, the clotting domain and inflammatory domain were not overlapping, was really the foundation of this invention was that we could maybe create this antibody to be able to target them in a selective way.Other Neurologic Conditions (22:31):So the antibody I developed is neutralizing blood toxicity by blocking the inflammatory domain of fibrin without adverse coagulation effects. And it's now completing phase one trials. So it has already completed the single ascending dose at 40 milligram per kilogram. It's interim data were announced already for this trial, with no safety signals. So if the antibody completes this year, the phase one trials, then it should be possible to be tested in different patient populations. You mentioned before chemo brain, and I think it's important that we think that blood-brain barrier disruption occurs among many neurological conditions, and it's an early event associated with early disease onset and worse prognosis in multiple sclerosis, Alzheimer's disease, traumatic injuries. So I think that it's by developing a strategy, therapeutic strategy to neutralize blood toxicity, this can have applications in a wide range of neurological conditions with vascular dysfunction.Eric Topol (23:54):Yeah, no. In your Nature Immunology 2020 piece [Figure below], you started with the 1883 identification of multiple sclerosis (MS) lesions were “engorged with blood”, the first link between blood leaks and brain inflammation. So this has enormous potential. And what I like about this Katerina is that you've dissected the clot component versus the inflammatory trigger of the fibrinogen and fibrin story. And this is so vital because if you keep throwing these things that just going to work on the clot and not deal with the pro-inflammatory consequences, then you're going to get the wrong impression that clots are not that important. And by the way, you did mention, and I want to come back to that too, endothelial inflammation, which is another feature of Long Covid is another kind of interactive part of this because when the lining of the blood vessel is inflamed, it will attract microthrombi and also be a participant in this whole affair. What do you think about Alzheimer's and the prospects of being able to interfere with Alzheimer's? We have 20 years in someone before this process takes hold and meets clinical manifestations. Would an antibody like this ever be useful along the way?Katerina Akassoglou (25:29):Yeah, so well, our antibody was tested first in Alzheimer's, this models when it was originally published, and we performed reversal trials in Alzheimer's models. So we dosed mice when they have established amyloid plaques, microglia activation, neuronal loss, and we could reverse this effect so it could increase cholinergic neurons in mice, reduce inflammation in a very selective way, only the neurotoxic part of inflammation and for genetic depletion of this pathway with akin mice in Alzheimer's disease. Also, improves from cognitive impairment, and we now have a new paper in Cell Press that is showing this effects also with really nice and unbiased machine learning models for behavioral segmentation [Figure below].So I think that there is the data both from genetic studies and the antibody show projection in Alzheimer's disease. And of course, as you might have read the recent Lancet report from the Lancet committee on dementia that identified the vascular risk factors as the key contributors, especially post sporadic cases of Alzheimer's disease that is over 90% of Alzheimer's disease that is not genetically linked.(26:58):So I think that there is a real need in Alzheimer's disease to be able to block this vascular induced pathology. And an antibody like the fibrin neutralizing therapy could be positioned to be protective from the vascular induced immune-mediated neurodegeneration in this disease as well. I mean, ultimately, I think that we need to be thinking the terms of efficacy. So we want to have a drug that is efficacious, but we also want it to be selective. And the selectivity is really important because the immune system has so many protective functions. So if we block phagocytosis, we end up with more debris, decrease of neurorepair, anti-myelination. So by blocking a ligand here and not blocking, not eliminating a cell type or blocking a global pathway in this cell, but biologic a single ligand, I think we have been able to achieve this balance between efficacy, but also safety because we only block this neurotoxic populations and not the entire innate immune response that also has been beneficial for metastatic functions in the brain.Blocking NeuroinflammationEric Topol (28:19):So you're bringing up another critical concept about targeting the inflammation, this kind of goldilocks story of how much you interfere with the immune response and how much you are able to reduce the adverse pro-inflammatory effects. So that gets me to what if we don't know in any given patient how much fibrin is having a role in their Long Covid. Although we know it has to be a prominent feature because we saw it in, not just a hospitalized patient series that I mentioned we reviewed, but other papers as well. But what about if you just try to take on inflammation like through a GLP-1 drug or cGAS–STING or any of these really strong anti-inflammatory pathways. Do you see a difference in a generalized approach versus a specific approach that is really fibrin centered?Katerina Akassoglou (29:22):Yeah, so we have a focus actually on both because we wanted to dissect the downstream intracellular pathways of fibrin, and it's interesting that we can find specific inflammatory mediators that potentially can also be targeted as well, to be able to preserve that specificity, which I think is really important because if we don't preserve the specificity, we'll end up with a lot of adverse effects by eliminating major immune responses. But the point that you raised I think is really important because it's not enough to have an efficacious and selective drug if you don't know the patient population that will benefit from this drug. So I think that in addition to the drug discovery studies, it's important to develop also biomarker programs with both fluid biomarkers, but also imaging biomarkers to be able to identify the patient populations that will benefit from such treatment.(30:25):So if for example, a patient population has a fibrin deposition, blocking only downstream might not be enough, and it might be really important to neutralize this fibrin toxicity in the brain of patients. And with our target engagement studies, we show that at least in animal models, the antibody can be there. So I'm very encouraged by also programs that are going on now in the scientific community to develop noninvasive ligands to be able to image fibrin in the brain that are already tested in different patient populations like multiple sclerosis. Because I think we're going to learn so much from the biology as we start interrogating and asking these questions now in different patient populations.Eric Topol (31:14):I think that's a vital point you're making because the success of a clinical trial here in a clinical syndrome that is mosaic with lots of different types of pathways. If you can nail down the patients that would have the most to stand to benefit from a particular intervention, that the chance of you not missing the benefit that is matching the marker, what image marker or other markers is so vital. Well, we've talked, I think, about some fascinating discoveries that you and your colleagues have made. I mean, it's really extraordinary, and obviously we need this in Long Covid. But you know what, Katerina, it's almost made me think that you were warming up to this for three decades, that somehow or other you were working on all this stuff and then came Covid. Is that how you see it, that somehow or other you didn't know that all the work you were doing was going to wind up in this space?Katerina Akassoglou (32:18):Oh, I never thought I would work in a virology project. This collaboration started over Zoom with Warner Greene. We were both sheltering in place. It was the beginning of the pandemic, and the first reports were coming out about this puzzling coagulopathy. And our labs were hardly operational at the time, as you know, we had to close down our labs for a while. And however, this was a very big problem, and we thought that this is our role as scientists. If we feel that we can contribute and we have the tools to contribute, we felt that it's important that we pivot some part of our research, and even we wouldn't be doing this before, but it was important to pivot a part of our research and collaborate. And I think studies like this, this study would have been impossible without a team of collaborators. As you know, there were over 50 scientists involved at Gladstone, UCSF, UCLA, UCSD, Stanford University. Without collaboration, this study wouldn't be possible. So I'm really grateful to everyone who came together to solve this problem because I think that's what scientists should be doing. We should be solving problems as they arise.Eric Topol (33:41):Well, and also, I think a lot of people don't realize that, for example, when the Covid vaccines came along, people think, oh, well, it all got done in 10 months since the sequence of the virus, when in fact it took 30 years at least between all the factors that went into having an mRNA and sequencing virus and nanoparticles. And in many ways, your arc of this work is like that because it took three decades to have all the tools and the basic understanding, the antibody that you had developed for different reasons and this fascinating unraveling of what's going on in the model and undoubtedly in some patients at least as well. So before we wrap up, have I missed anything about this just remarkable work you've done?Katerina Akassoglou (34:33):Oh, thank you. I just want to thank you for this discussion and thank you for emphasizing the different areas and the different decisions that this pathway can have implications both for our understanding, our basic understanding of the blood brain immune interface, as well as also potential translation. And I think that the curiosity sometimes of how things work, I never thought it would work on Covid, like you mentioned at the beginning, but I think that basic science and curiosity driven science can sometimes lead to discoveries with translational implications that hopefully might benefit patients one day.Eric Topol (35:21):Yeah, well, undoubtedly it will. We're indebted to you, Katerina and all the folks that you have teamed up with, connecting the dots at the neurovascular interface. Phenomenal work and will follow the subsequent with great interest and it will likely not just a story about Long Covid, but other areas as well, so thank you.*********************************Thanks for listening, reading or watching!The Ground Truths newsletters and podcasts are all free, open-access, without ads.Please share this post/podcast with your friends and network if you found it informative!Voluntary paid subscriptions all go to support Scripps Research. Many thanks for that—they greatly help fund our summer internship programs.Thanks to my producer Jessica Nguyen and Sinjun Balabanoff for audio and video support at Scripps Research.Note: you can select preferences to receive emails about newsletters, podcasts, or all I don't want to bother you with an email for content that you're not interested in. Get full access to Ground Truths at erictopol.substack.com/subscribe

Meet Dr. Shivani Gupta, entrepreneur, and PhD in Turmeric. Today we discuss the benefits of Turmeric, Ayurvedic medicine, and how this amazing plant and curcumin extra can help with pain management. Here are some scientific studies citing the benefits of turmeric in pain management like sciatica:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3946321/These results indicated that curcumin exerted a therapeutic role in neuropathic pain by down-regulating p300/CBP HAT activity-mediated gene expression of BDNF and Cox-2https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7600446/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10610406/CUR decreased CD11b and GFAP gene expression in the spinal cord. BDMC decreased IBA-1 in the spinal cord and amygdala as well as CD11b and GFAP in the spinal cord. Both CUR and BDMC reduced PGC1α gene expression in the amygdala, PINK1 gene expression in the spinal cord, and TLR4 in the spinal cord and amygdala, while they increased Complex I and SOD1 gene expression in the spinal cord. CUR and BDMC administration decreased mechanical hypersensitivity in NP by mitigating glial activation, oxidative stress, and mitochondrial dysfunction.You can hear more from Dr. Shivani at shivanigupta.com and check out her other site: https://fusionaryformulas.com/ (affiliate link)You can use “sciatica” to get 15% off your order!Are you looking for a more affordable way to manage your pain? Check out the patient advocate program here: ptpatientadvocate.comHere's the self cheat sheet for symptom management: https://ifixyoursciatica.gymleadmachine.co/self-treatment-cheat-sheet-8707Book a free strategy call: https://msgsndr.com/widget/appointment/ifixyoursciatica/strategy-callSupport this podcast at — https://redcircle.com/fix-your-sciatica-podcast/donationsAdvertising Inquiries: https://redcircle.com/brandsPrivacy & Opt-Out: https://redcircle.com/privacy

Too busy to read The Lens? Listen to our weekly summary here! In this week's episode: A retrospective study reveals demographic risk factors for open globe injuries requiring repairs and poorer visual outcomes. Mapping of access to laser eye surgery in 5 Midwest states did not reveal shorter travel times despite expansion of surgical privileges to optometrists. Twice-daily pilocarpine for presbyopia treatment showed greater improvements in distance-corrected near visual acuity compared to once-daily dosing Inhibition of CD11b, a component of the complement system, may provide a neuroprotective effect in optic nerve injury.

Too busy to read the Lens? Listen to our weekly summary here! In these week issue: A port delivery system with ranibizumab shows non-inferiority compared to monthly ranibizumab injections Treatment with low dose atropine nightly for two years shows no difference in spherical equivalent or axial length compared to placebo A new mobility outcome (MOST, MObility Standardized Test) is validated in measuring disease progression in patients with retinitis pigmentosa In a mouse model of traumatic optic neuropathy, CD11b is significantly elevated in microglia near the site of injury

Here’s why eating garlic and onions can prevent hypertension and diabetes Federal University of Technology (Nigeria), April 16, 2021 n a recent study, researchers at the Federal University of Technology in Nigeria investigated the benefits of eating garlic, white onion and purple onion against serious conditions like diabetes and hypertension. They confirmed these by looking at how extracts from the three alliums affect the activity of diabetes-related enzymes, such as a-amylase and a-glucosidase, and the hypertension-related enzyme, angiotensin-converting enzyme (ACE). The researchers reported their findings in an article published in the Journal of Dietary Supplements. Garlic, white onion and purple onion show antioxidant, antidiabetic and antihypertensive properties Garlic and onions are spices commonly used in cooking. They also serve as ingredients in several traditional delicacies in Nigeria that are known to contain plenty of polyphenols. To assess the beneficial properties of garlic, white onion and purple onion, the researchers first obtained extracts from each and assessed their inhibitory effects on certain enzymes. They also conducted assays to determine the antioxidant capacities of the extracts. ACE is the enzyme responsible for converting angiotensin I into angiotensin II, the hormone that increases blood pressure, as well as body water and sodium content. Angiotensin II elevates blood pressure by constricting the blood vessels; hence, chemicals that can inhibit the activity of ACE, which is responsible for the production of angiotensin II, are used for the treatment of hypertension. (Related: Meet the “two-day cure” plant: An African medicinal plant that can naturally lower blood pressure.) a-Amylase is the enzyme that breaks down starch and glycogen into glucose and maltose (two glucose molecules bound together). In humans, this enzyme is produced by the salivary glands and the pancreas. a-Glucosidase, on the other hand, is responsible for breaking down carbohydrates in the small intestine and facilitating the absorption of glucose. Inhibiting the activity of this enzyme is one of the strategies currently used to prevent the rise of blood sugar levels following a carbohydrate-filled meal. The researchers reported that the garlic, purple onion and white onion extracts inhibited the activities of ACE, a-amylase and a-glucosidase in vitro in a concentration-dependent manner. At a half maximal inhibitory concentration (IC50) of 0.59 mg/mL, the purple onion extract exhibited a higher inhibitory effect on ACE than the white onion extract (IC50 = 0.66 mg/mL) and the garlic (IC50 = 0.96 mg/mL) extract. Meanwhile, the white onion extract showed a significantly stronger inhibitory effect on a-amylase at an IC50 of 3.93 mg/mL than the garlic extract (IC50 = 8.19 mg/mL) and the purple onion (IC50 = 8.27 mg/mL) extract. The garlic extract, on the other hand, showed a similar inhibitory effect (IC50 = 4.50 mg/mL) on a-glucosidase as the white and purple onion extracts. All three extracts also showed dose-dependent free radical scavenging activity and reducing power in the antioxidant assays. Based on these findings, the researchers concluded that garlic, white onion and purple onion can be used to treat or prevent diabetes and hypertension, thanks to their ability to inhibit ACE, a-amylase and a-glucosidase activity, as well as lipid peroxidation in the pancreas and the heart.   Adolescents with lack of empathy show early signs of psychopathy   University of Coimbra (Portugal), April 14, 2021 A pioneering study with the Portuguese population shows that adolescents with high levels of callous-unemotional traits demonstrate lower levels of anticipated guilt towards the possibility of committing an immoral act and struggle to judge an immoral act as a wrong one. Researchers have evaluated the callous traits, that is, the lack of empathy and disregard for the wellbeing and feelings of others, of 47 adolescents between 15 and 18 years old. The teenagers watched video animations portraying examples of moral transgressions, such as incriminating someone or keeping money that fell from someone else's pocket. "This approach allowed us to create more realistic scenarios that happen in daily life," explains Oscar Gonçalves, a neuroscientist at Proaction Lab and co-author of the study. The adolescents were asked how guilty they would feel if they were the ones to commit the moral transgressions and how wrong they think the actions were.  Although the callous-unemotional traits in adolescents are known to be precursors of psychopathy in adulthood, the results of the study differ from what is known about psychopaths. "Adults with psychopathic traits show low levels of anticipated guilt but consider immoral actions as wrong. However, in our study, adolescents with high CU levels show levels of guilt and judge immoral actions as less wrong," explains Margarida Vasconcelos, first author.  However, researchers have found evidence of a dissociation between moral emotions and moral judgment, that is, between the feelings of guilt and the judgment of immoral actions. "Even in adolescents with sub-clinical levels of callous-unemotional traits, this dissociation typical in psychopathy in adulthood is already happening during development," explains the study coordinator Ana Seara Cardoso.  The results of the study will "contribute to the development of a severe anti-social behavior model" and allow the "development of intervention targets, rehabilitation and early prevention of anti-social behavior," says Ana Seara Cardoso.   Omega-3 supplements do double duty in protecting against stress   Ohio State University, April 20, 2021 A high daily dose of an omega-3 supplement may help slow the effects of aging by suppressing damage and boosting protection at the cellular level during and after a stressful event, new research suggests. Researchers at The Ohio State University found that daily supplements that contained 2.5 grams of omega-3 polyunsaturated fatty acids, the highest dose tested, were the best at helping the body resist the damaging effects of stress. Compared to the placebo group, participants taking omega-3 supplements produced less of the stress hormone cortisol and lower levels of a pro-inflammatory protein during a stressful event in the lab. And while levels of protective compounds sharply declined in the placebo group after the stressor, there were no such decreases detected in people taking omega-3s. The supplements contributed to what the researchers call stress resilience: reduction of harm during stress and, after acute stress, sustained anti-inflammatory activity and protection of cell components that shrink as a consequence of aging. The potential anti-aging effects were considered particularly striking because they occurred in people who were healthy but also sedentary, overweight and middle-aged—all characteristics that could lead to a higher risk for accelerated aging. "The findings suggest that omega-3 supplementation is one relatively simple change people could make that could have a positive effect at breaking the chain between stress and negative health effects," said Annelise Madison, lead author of the paper and a graduate student in clinical psychology at Ohio State. The research is published today (Monday, April 19, 2021) in the journal Molecular Psychiatry. Madison works in the lab of Janice Kiecolt-Glaser, professor of psychiatry and psychology and director of the Institute for Behavioral Medicine Research at Ohio State. This paper is a secondary analysis of one of Kiecolt-Glaser's earlier studies showing that omega-3 supplements altered a ratio of fatty acid consumption in a way that helped preserve tiny segments of DNA in white blood cells. Those short fragments of DNA are called telomeres, which function as protective caps at the end of chromosomes. Telomeres' tendency to shorten in many types of cells is associated with age-related diseases, especially heart disease, and early mortality. In the initial study, researchers were monitoring changes to telomere length in white blood cells known as lymphocytes. For this new study, the researchers looked at how sudden stress affected a group of biological markers that included telomerase, an enzyme that rebuilds telomeres, because levels of the enzyme would react more quickly to stress than the length of telomeres themselves. Specifically, they compared how moderate and high doses of omega-3s and a placebo influenced those markers during and after an experimental stressor. Study participants took either 2.5 grams or 1.25 grams of omega-3s each day, or a placebo containing a mix of oils representing a typical American's daily intake. After four months on the supplements, the 138 research participants, age 40-85, took a 20-minute test combining a speech and a math subtraction task that is known to reliably produce an inflammatory stress response. Only the highest dose of omega-3s helped suppress damage during the stressful event when compared to the placebo group, lowering cortisol and a pro-inflammatory protein by an average of 19% and 33%, respectively. Results from blood samples showed that both doses of omega-3s prevented any changes in telomerase levels or a protein that reduces inflammation in the two hours after participants experienced the acute stress, meaning any needed stress-related cell repair—including telomere restoration—could be performed as usual. In the placebo group, those repair mechanisms lost ground: Telomerase dropped by an average of 24% and the anti-inflammatory protein decreased by an average of at least 20%. "You could consider an increase in cortisol and inflammation potential factors that would erode telomere length," Madison said. "The assumption based on past work is that telomerase can help rebuild telomere length, and you want to have enough telomerase present to compensate for any stress-related damage. "The fact that our results were dose-dependent, and we're seeing more impact with the higher omega-3 dose, would suggest that this supports a causal relationship." The researchers also suggested that by lowering stress-related inflammation, omega-3s may help disrupt the connection between repeated stress and depressive symptoms. Previous research has suggested that people with a higher inflammatory reaction to a stressor in the lab may develop more depressive symptoms over time. "Not everyone who is depressed has heightened inflammation—about a third do. This helps explain why omega-3 supplementation doesn't always result in reduced depressive symptoms," Kiecolt-Glaser said. "If you don't have heightened inflammation, then omega-3s may not be particularly helpful. But for people with depression who do, our results suggest omega-3s would be more useful." The 2.5-gram dose of omega-3s is much higher than what most Americans consume on a daily basis, but study participants showed no signs of having problems with the supplements, Madison said.           Want to be robust at 40-plus? Meeting minimum exercise guidelines won't cut it 5 hours of moderate activity a week may be required to avoid midlife hypertension, UCSF-led study shows University of California at San Francisco, April 15, 2021 Young adults must step up their exercise routines to reduce their chances of developing high blood pressure or hypertension - a condition that may lead to heart attack and stroke, as well as dementia in later life. Current guidelines indicate that adults should have a minimum of two-and-a-half hours of moderate intensity exercise each week, but a new study led by UCSF Benioff Children's Hospitals reveals that boosting exercise to as much as five hours a week may protect against hypertension in midlife - particularly if it is sustained in one's thirties, forties and fifties.  In the study publishing in American Journal of Preventive Medicine on April 15, researchers followed approximately 5,000 adults ages 18 to 30 for 30 years. The participants were asked about their exercise habits, medical history, smoking status and alcohol use. Blood pressure and weight were monitored, together with cholesterol and triglycerides.  Hypertension was noted if blood pressure was 130 over 80 mmHg, the threshold established in 2017 by the American College of Cardiology/American Heart Association.  The 5,115 participants had been enrolled by the Coronary Artery Risk Development in Young Adults (CARDIA) study and came from urban sites in Birmingham, Ala., Chicago, Minneapolis and Oakland, Calif. Approximately half the participants were Black (51.6 percent) and the remainder were White. Just under half (45.5 percent) were men.  Fitness Levels Fall Fast for Black Men Leading to More Hypertension Among the four groups, who were categorized by race and gender, Black men were found to be the most active in early adulthood, exercising slightly more than White men and significantly more than Black women and White women. But by the time Black men reached age 60, exercise intake had slumped from a peak of approximately 560 exercise units to around 300 units, the equivalent to the minimum of two-and-a-half hours a week of moderate intensity exercise recommended by the U.S. Department of Health and Human Services. This was substantially less exercise than White men (approximately 430 units) and slightly more than White women (approximately 320 units). Of the four groups, Black women had the least exercise throughout the study period and saw declines over time to approximately 200 units. "Although Black male youth may have high engagement in sports, socio-economic factors, neighborhood environments, and work or family responsibilities may prevent continued engagement in physical activity through adulthood," said first author Jason Nagata, MD, of the UCSF Division of Adolescent and Young Adult Medicine. Additionally, Black men reported the highest rates of smoking, which may preclude physical activity over time, he noted. Physical activity for White men declined in their twenties and thirties and stabilized at around age 40. For White women, physical activity hovered around 380 exercise units, dipping in their thirties and remaining constant to age 60. Rates of hypertension mirrored this declining physical activity. Approximately 80-to-90 percent of Black men and women had hypertension by age 60, compared with just below 70 percent for White men and 50 percent for White women.  "Results from randomized controlled trials and observational studies have shown that exercise lowers blood pressure, suggesting that it may be important to focus on exercise as a way to lower blood pressure in all adults as they approach middle age," said senior author Kirsten Bibbins-Domingo, MD, PhD, of the UCSF Department of Epidemiology and Biostatistics.  "Teenagers and those in their early twenties may be physically active but these patterns change with age. Our study suggests that maintaining physical activity during young adulthood - at higher levels than previously recommended - may be particularly important." More Exercise from Youth to Midlife Offers Best Protection Against Hypertension When researchers looked at the 17.9 percent of participants who had moderate exercise for at least five hours a week during early adulthood - double the recommended minimum - they found that the likelihood of developing hypertension was 18 percent lower than for those who exercised less than five hours a week. The likelihood was even lower for the 11.7 percent of participants who maintained their exercise habits until age 60.  Patients should be asked about physical activity in the same way as they are routinely checked for blood pressure, glucose and lipid profiles, obesity and smoking, Nagata said, and intervention programs should be held at schools, colleges, churches, workplaces and community organizations. Black women have high rates of obesity and smoking, and low rates of physical activity, he said, and should be an important group for targeted intervention.  "Nearly half of our participants in young adulthood had suboptimal levels of physical activity, which was significantly associated with the onset of hypertension, indicating that we need to raise the minimum standard for physical activity," Nagata said. "This might be especially the case after high school when opportunities for physical activity diminish as young adults transition to college, the workforce and parenthood, and leisure time is eroded."     Study finds association between periodontal disease and low intake of minerals, vitamins and dietary fiber in young adult women Tokyo Medical and Dental University, April 12, 2021 According to news reporting out of Tokyo, Japan, research stated, “Dietary habits of middle-aged and elderly individuals affected by periodontal disease (PD) differ from those who are unaffected by it, according to previous reports. However, in young adults, there are only a few reports that show a correlation between nutrient/food intake and PD.” Our news journalists obtained a quote from the research from Tokyo Medical and Dental University (TMDU), “Moreover, no report till date has assessed the correlation between dietary habits and PD using a self-administered diet history questionnaire (DHQ). Therefore, we assessed this correlation using a DHQ in young adult women who are likely to develop PD. The participants were enrolled from 2 universities and included 120 female college students a mean age of 20.4 y. The participants were assessed for the presence of PD according to the community periodontal index and were divided into two groups, the PD group and the non-PD group. Their dietary habits were investigated using a DHQ and the level of difficulty in chewing food was assessed. The PD group had a significantly lower nutrient intake of minerals, fat-soluble vitamins, water-soluble vitamins, and dietary fiber than the non-PD group. In terms of food groups, the PD group consumed significantly lesser amounts of green and yellow vegetables (GYV) than the non-PD group. Multivariate analysis revealed that the PD group had significantly lower intakes of vitamin E and GYV than the non-PD group. The PD group consumed significantly lesser amounts of hard foods than the non-PD group.” According to the news editors, the research concluded: “Young adult women who were evaluated for PD by a screening test had a significantly lower nutrient/food intake than those without a PD.” This research has been peer-reviewed.       Just 2 days of increased sugar intake can harm your gut health, warn researchers University of Alberta, April 16, 2021 Researchers from the University of Alberta in Canada found that short-term increases in sugar intake can increase the risk of inflammatory bowel disease. Their finding, which was published in Scientific Reports, is a reminder that eating healthy must be sustained in order to keep your gut in good shape. “Surprisingly, our study shows that short-term sugar consumption can really have a detrimental impact, and so this idea that it’s OK to eat well all week and indulge in junk food on the weekend is flawed,” said Karen Madsen, one of the study researchers. Increased sugar intake is bad for the gut Previous studies have shown that diets can affect your susceptibility to disease. Western diets, for example, have been implicated in the development of inflammatory bowel disease. But it’s still unclear when a poor diet begins to take a toll on your health, much less how it does so. To investigate, the researchers placed adult mice on a chow diet or a high-sugar diet and treated them with dextran sodium sulfate to induce ulcerative colitis, one of the major forms of inflammatory bowel disease. Disease severity was assessed daily. After two days, the mice on the high-sugar diet were at great risk of developing colitis. Their immune response also weakened while their gut permeability increased, allowing more bacteria and toxins to enter their bloodstream. “We wanted to know how long it takes before a change in diet translates into an impact on health. In the case of sugar and colitis, it only took two days, which was really surprising to us. We didn’t think it would happen so quickly,” said Madsen. The researchers attributed these effects to sugar’s impact on the gut bacteria. Eating sugary foods decreases the amount of “good” gut bacteria that produce short-chain fatty acids, which are critical for a strong immune response. Meanwhile, sugar feeds “bad” bacteria that promote inflammation and weaken your immunity.  Fortunately, the researchers found that supplementing with short-chain fatty acids helped reduce the negative effects of a high-sugar diet. Having these supplements as an option will be great for people struggling to change their bad eating habits. “People want to eat what they want to eat, so short-chain fatty acids could possibly be used as supplements to help protect people against the detrimental effects of sugar on inflammatory bowel disease,” said Madsen.     Rose water is an antimicrobial and anti-inflammatory remedy for skin infections Teikyo University (Japan), April 15, 2021 Rosa damascena, commonly known as Damask rose, is one of the most important and medicinally useful members of the Rosaceae (rose) family. It is an ornamental plant widely used to make perfumes and is reported to have plenty of beneficial properties. According to multiple studies, Damask rose has anti-HIV, antibacterial, antioxidant, antitussive, hypnotic and antidiabetic properties. It has also shown relaxant effects on the tracheal chains of guinea pigs. In a recent study, researchers at Teikyo University in Japan investigated two biological properties of Damask rose, specifically it’s antimicrobial and anti-inflammatory properties. They tested rose water made from high-quality Damask rose petals on two microbial pathogens, namely, Candida albicans and methicillin-resistant Staphylococcus aureus (MRSA), which commonly cause skin infections. The researchers reported their findings in an article published in Biological and Pharmaceutical Bulletin. Damask rose water is a natural antibiotic and anti-inflammatory agent Damask rose is a multipurpose plant widely known for its culinary and medicinal applications, among other things. Edible parts of Damask rose are used in various cuisines, including its young shoots, petals, fruits, leaves and seeds. Damask rose petals are used to make jams and add flavor to beverages, baked goods and desserts. They are also used for cooking dishes. Rosewater, which can be sweetened to produce rose syrup, is a byproduct of rose oil production. It is usually obtained by steam distilling Damask rose petals and taking the hydrosol portion of the rose petal distillate.  In different parts of the world, rose water, rose oil and a decoction made of Damask rose roots are used in traditional medicine for the treatment of various ailments, such as abdominal and chest pain, digestive problems and inflammation, especially of the neck. In North America, Indian tribes use the decoction as a cough remedy for children. Rose oil is used to treat depression and reduce stress and tension. Inhaling the vapor produced by heating rose oil is also believed to be an effective remedy for allergies, headaches and migraine. Damask rose water, on the other hand, is traditionally used to treat skin conditions, such as erythema (skin redness), itchiness and swelling. To evaluate its antimicrobial and anti-inflammatory properties, the researchers tested Damask rose water against C. albicans and MRSA and assessed its effects on the function of neutrophils, which are white blood cells that serve as key regulators of inflammatory reactions. The researchers reported that Damask rose water (2.2. percent solution) inhibited the mycelial growth of C. albicans and reduced the viability of MRSA within an hour of treatment. Damask rose water (five to 15 percent) also suppressed the activation of neutrophils induced by treatment with lipopolysaccharide (LPS), a bacterial toxin; tumor necrosis factor-alpha (TNF-a), a cell-signaling protein produced by immune cells; and N-formyl-Met-Leu-Phe (fMLP), a macrophage activator. Additionally, Damask rose water reduced LPS- and TNF-a-induced cell surface expression of the adhesion-related molecule, cluster of differentiation 11b (CD11b), which is rapidly elevated by the activation of neutrophils. The amount of CD11b in neutrophils is said to correlate with their activation and inflammation. However, Damask rose water did not affect the migratory capacity of neutrophils (with or without a chemoattractant). Based on these findings, the researchers concluded that Damask rose water can reduce the pathogenicity of microbes and attenuate neutrophil stimulation, thus inhibiting skin inflammation caused by microbial infections.     Study shows how chronic stress may inhibit the body's cancer-fighting ability University of Western Ontario, April 15, 2021 New research from Western University has shown how psychological stress hinders the immune system's defenses against cancer. By investigating the effects of chronic stresson the immune system's "emergency responders," researchers at the Schulich School of Medicine & Dentistry found that a stress-induced hormone impairs the ability of these immune cells to carry out their cancer-fighting function. Led by Mansour Haeryfar, Ph.D., the research looked specifically at innate-like T cells, which when functioning properly enable the immune system to look for potentially cancerous cells in the body and destroy them. The study was published today in Cell Reports. Innate-like T cells include invariant natural killer T (iNKT) and mucosa-associated invariant T (MAIT) cells, which were the subjects of this investigation. iNKT cells are present in small numbers in many tissues but are especially enriched in the human omentum, an apron-like layer of fatty tissue. MAIT cells are present in relatively high numbers in the human peripheral blood, gut, lungs and liver among other organs. "These innate-like T cells are our immune system's emergency responders," said Haeryfar. "They react quickly to pathogens and cancer cells and are in a pre-activated mode, so they are like loaded guns, ready to respond." Previous studies have shown that when a person experiences chronic psychological and emotional stress, the body's immune system is suppressed, dampening its ability to fight cancer and opportunistic infections. This happens in large part because stress hormones kill off some of the body's immune cells. However, Haeryfar and his team showed that innate-like T cells actually don't die as a result of chronic stress but their cancer-fighting abilities are drastically impaired by stress-induced hormones called glucocorticoids. This impairment led to a striking increase in cancer metastasis in a mouse model. "We found that innate-like T cells survive when the host is under stress, but their functions are compromised," Haeryfar said. "The cells cannot make enough of their beneficial mediators to help fight cancer, so the metastatic burden is increased because of the stress." The team also looked at the effects of natural and synthetic glucocorticoids on innate-like T cells in human blood and liver tissue, where they are abundant. This was important to providing initial evidence that some of the discoveries made in the mouse models were valid for human cells as well, said Patrick Rudak, Ph.D. Candidate in Haeryfar's lab. One of the important implications of this work is that innate-like T cells are currently being investigated for cancer immunotherapy treatment. This study demonstrates that their therapeutic potential can be dampened by psychological stress, said Haeryfar, and this finding needs to be considered when designing or administering those therapies. Rudak added: "Our study demonstrates that, despite being capable of instigating robust anti-tumor immune responses under normal conditions, innate-like T cells completely fail to protect against tumors during psychological stress." Because the study also uncovered the mechanisms by which stress diminishes T cell function, the researchers hope they can use the information to help design immunotherapies involving these cells that will still be effective in psychologically stressed patients.

The TLR7 agonist imiquimod has been used successfully as adjuvant for skin treatment of virus-associated warts and basal cell carcinoma. The effects of skin TLR7 triggering on respiratory leukocyte populations are unknown. In a placebo-controlled experimental animal study we have used multicolour flow cytometry to systematically analyze the modulation of respiratory leukocyte subsets after skin administration of imiquimod. Compared to placebo, skin administration of imiquimod significantly increased respiratory dendritic cells (DC) and natural killer cells, whereas total respiratory leukocyte, alveolar macrophages, classical CD4+ T helper and CD8+ T killer cell numbers were not or only moderately affected. DC subpopulation analyses revealed that elevation of respiratory DC was caused by an increase of respiratory monocytic DC and CD11b(hi) DC subsets. Lymphocyte subpopulation analyses indicated a marked elevation of respiratory natural killer cells and a significant reduction of B lymphocytes. Analysis of cytokine responses of respiratory leukocytes after stimulation with Klebsiella pneumonia indicated reduced IFN-γ and TNF-α expression and increased IL-10 and IL-12p70 production after 7 day low dose skin TLR7 triggering. Additionally, respiratory NK cytotoxic activity was increased after 7d skin TLR7 triggering. In contrast, lung histology and bronchoalveolar cell counts were not affected suggesting that skin TLR7 stimulation modulated respiratory leukocyte composition without inducing overt pulmonary inflammation. These data suggest the possibility to modulate respiratory leukocyte composition and respiratory cytokine responses against pathogens like Klebsiella pneumonia through skin administration of a clinically approved TLR7 ligand. Skin administration of synthetic TLR7 ligands may represent a novel, noninvasive means to modulate respiratory immunity.

The NF-kappa B/REL-family of transcription factors plays a central role in coordinating the expression of a wide variety of genes controlling immune responses including autoimmunity of the central nervous system (CNS). The inactive form of NF-kappa B consists of a heterodimer which is complexed with its inhibitor, I kappa B. Conditional knockout-mice for I kappa B alpha in myeloid cells (IysMCreI kappa B alpha(fl/fl)) have been generated and are characterized by a constitutive activation of NF-kappa B proteins allowing the study of this transcription factor in myelin-oligodendrocyte-glycoprotein induced experimental autoimmune encephalomyelitis (MOG-EAE), a well established experimental model for autoimmune demyelination of the CNS. In comparison to controls, IysMCreI kappa B alpha(fl/fl) mice developed a more severe clinical course of EAE. Upon histological analysis on day 15 p.i., there was an over two fold increased infiltration of T-cells and macrophages/microglia. In addition, IysMCreI kappa B alpha(fl/fl) mice displayed an increased expression of the NF-kappa B dependent factor inducible nitric oxide synthase in inflamed lesions. These changes in the CNS are associated with increased numbers of CD11b positive splenocytes and a higher expression of Ly6c on monocytes in the periphery. Well in accordance with these changes in the myeloid cell compartment, there was an increased production of the monocyte cytokines interleukin(IL)-12 p70, IL-6 and IL-1beta in splenocytes. In contrast, production of the T-cell associated cytokines interferon gamma (IFN-gamma) and IL-17 was not influenced. In summary, myeloid cell derived NF-kappa B plays a crucial role in autoimmune inflammation of the CNS and drives a pathogenic role of monocytes and macrophages independently from T-cells.

Background: Clinical trials evaluating anti-CD20-mediated B-cell depletion in multiple sclerosis (MS) and neuromyelitis optica (NMO) generated encouraging results. Our recent studies in the MS model experimental autoimmune encephalomyelitis (EAE) attributed clinical benefit to extinction of activated B-cells, but cautioned that depletion of naive B-cells may be undesirable. We elucidated the regulatory role of un-activated B-cells in EAE and investigated whether anti-CD20 may collaterally diminish regulatory B-cell properties in treatment of neuroimmunological disorders. Methods: Myelin oligodendrocyte glycoprotein (MOG) peptide-immunized C57Bl/6 mice were depleted of B-cells. Functional consequences for regulatory T-cells (Treg) and cytokine production of CD11b(+) antigen presenting cells (APC) were assessed. Peripheral blood mononuclear cells from 22 patients receiving anti-CD20 and 23 untreated neuroimmunological patients were evaluated for frequencies of B-cells, T-cells and monocytes; monocytic reactivity was determined by TNF-production and expression of signalling lymphocytic activation molecule (SLAM). Results: We observed that EAE-exacerbation upon depletion of un-activated B-cells closely correlated with an enhanced production of pro-inflammatory TNF by CD11b(+) APC. Paralleling this pre-clinical finding, anti-CD20 treatment of human neuroimmunological disorders increased the relative frequency of monocytes and accentuated pro-inflammatory monocyte function; when reactivated ex vivo, a higher frequency of monocytes from B-cell depleted patients produced TNF and expressed the activation marker SLAM. Conclusions: These data suggest that in neuroimmunological disorders, pro-inflammatory APC activity is controlled by a subset of B-cells which is eliminated concomitantly upon anti-CD20 treatment. While this observation does not conflict with the general concept of B-cell depletion in human autoimmunity, it implies that its safety and effectiveness may further advance by selectively targeting pathogenic B-cell function.

Background: Clinical presentation, diagnosis, management and outcome of molecularly defined congenital pulmonary alveolar proteinosis (PAP) due to mutations in the GM-CSF receptor are not well known. Case presentation: A 2 1/2 years old girl was diagnosed as having alveolar proteinosis. Whole lung lavages were performed with a new catheter balloon technique, feasible in small sized airways. Because of some interstitial inflammation in the lung biopsy and to further improve the condition, empirical therapy with systemic steroids and azathioprin, and inhaled and subcutaneous GMCSF, were used. Based on clinical measures, total protein and lipid recovered by whole lung lavages, all these treatments were without benefit. Conversely, severe respiratory viral infections and an invasive aspergillosis with aspergilloma formation occurred. Recently the novel homozygous stop mutation p.Ser25X of the GMCSF receptor alpha chain was identified in the patient. This mutation leads to a lack of functional GMCSF receptor and a reduced response to GMCSF stimulation of CD11b expression of mononuclear cells of the patient. Subsequently a very intense treatment with monthly lavages was initiated, resulting for the first time in complete resolution of partial respiratory insufficiency and a significant improvement of the overall somato-psychosocial condition of the child. Conclusions: The long term management from early childhood into young adolescence of severe alveolar proteinosis due to GMCSF receptor deficiency requires a dedicated specialized team to perform technically demanding whole lung lavages and cope with complications.

Die equine rezidivierende Uveitis (ERU) ist eine spontane, periodisch wiederkehrende Erkrankung und ist die häufigste Ursache von Erblindung bei adulten Pferden. Sie betrifft weltweit bis zu 10% der Pferdepopulation und ist das einzige spontane Modell für die autoimmun-mediierte Uveitis des Menschen. Ziel dieser Arbeit war es, mittels vergleichender Proteomanalyse Unterschiede in der Proteinexpression zwischen Leukozyten gesunder und an ERU erkrankter Pferde zu detektieren und durch massenspektrometrische Identifizierung dieser differenziell exprimierten Proteine die Pathogenese der ERU weiter zu charakterisieren. In diesem Zusammenhang wurden in dieser Studie Leukozyten aus dem peripheren Blut von insgesamt 37 augengesunden und 33 an ERU erkrankten Pferden untersucht. Mittels zweidimensionaler Difference-gel-electrophoresis (2D-DIGE) wurden in einem ersten, lymphozytären Experiment 50 differenziell exprimierte Proteine detektiert. Von diesen Proteinen konnten insgesamt neun massenspektrometrisch eindeutig identifiziert werden, unter anderem Septin 7. Die Expressionsstärke von Septin 7 in Lymphozyten von an ERU erkrankten Pferden ist im Vergleich zu gesunden Tieren um 30% verringert. Zudem ist die Septin 7-Expression in T-Zellen der bei ERU intraokulär gebildeten Lymphfollikel niedriger als in den Zellen des als Kontrollgewebe eingesetzten Mandibularlymphknotens. Nachdem Septin 7 eine Rolle bei der Funktion und Migration von T-Zellen zugesprochen wird, soll in weiteren Untersuchungen geklärt werden, wie die verringerte Septin 7-Expression mit der Pathologie der ERU zusammenhängen könnte. Im zweiten Experiment wurden auch die Granulozyten in die Analyse mit einbezogen. Aus den hier ebenfalls 50 unterschiedlich abundanten Proteinen konnten 20 eindeutig identifiziert werden, darunter Talin 1, das in den ERU-Proben zu 89% niedriger exprimiert war als in den Kontrollen. Talin 1 wird auf equinen PBL zum Teil mit mehreren Integrinen (CD11a, CD11b, CD29 und CD49a) ko-exprimiert und war auf den intraokulären Zellen zu nahezu 100% exprimiert im Vergleich zum Kontroll-Lymphknoten (ca. 40%). Da Talin 1 ebenfalls eine Rolle bei der Zellmigration zugeschrieben wird, sollen weitere funktionelle Analysen klären, welche Bedeutung die veränderte Talin 1-Expression in den PBL und den intraokulären Leukozyten der an ERU erkrankten Pferde hat. Weitere interessante Kandidaten aus beiden Experimenten, die unserer Meinung nach nähere Untersuchungen rechtfertigen, sind Scaffold attachment Faktor B, Superoxid Dismutase und Triosephosphat Isomerase (überexprimiert bei ERU) sowie Programmed cell death 6-interagierendes Protein, NCK Adapter Protein 1 und Ezrin (niedriger exprimiert bei ERU).

Background: Induced sputum is the most commonly used method to analyze airway inflammation in cystic fibrosis (CF) patients ex vivo. Due to the complex matrix of the sample material, precise and reliable analysis of sputum constituents depends critically on preanalytical issues. Objectives: Here we compared the commonly used method for sputum processing by dithiothreitol (DTT) with a novel mechanical method in regard to basal cellular parameters, neutrophil markers and glutathione (GSH) levels. Methods: Sputum samples from CF patients were processed in parallel with or without the use of DTT. The key improvement of the mechanical method was the processing in many very small aliquots. Cellular and humoral markers were assessed and compared according to Bland-Altman. Results: Total cell count, cell viability, differential cell count, neutrophil elastase levels and flow cytometrically analyzed neutrophil markers (CD63, CD11b, DHR) did not differ between the two methods. Intracellular and extracellular GSH levels were significantly higher in DTT-treated samples (p = 0.002). Conclusion: The mechanical sputum-processing method presented had a similar yield of cells and fluids as the conventional DTT method and the advantage of omitting the introduction of reducing agents. This method allows a more reliable analysis of redox-dependent airway inflammation in sputum cells and fluid from CF patients than methods utilizing DTT. Copyright (C) 2009 S. Karger AG, Basel

Neben den klassischen kardiovaskulären Risikofaktoren wie arterielle Hypertonie oder Hypercholesterinämie kommen den psychosozialen Faktoren wie Stress oder Depression eine entscheidene Rolle als Risikofaktor für die Entwicklung der Atherosklerose zu. Obwohl das chronische Stresshormon Corticotropin-Releasing-Hormon im Rahmen der adaptiven Stressantwort als Hauptvertreter der Effektorhormone angesehen wird, sind die pathophysiologischen Mechanismen, die zu einer CRH/Stress-bedingten endothelialen Dysfunktion führen, weitgehend unbekannt. Diese Arbeit hatte zum Ziel, den Effekt von peripherem CRH auf die Monozyten/Endothel-Interaktion, beispielhaft die Adhäsion, herauszuarbeiten. Die Untersuchungen der Monozyten-Endothel-Adhäsion wurde in einem in-vitro-Modell unter Verwendung der Zelllinien HMEC-1 und THP-1 mit einer neuen, modifizierten fluorometrischen Methode untersucht, monozytäres MAC-1/CD11b, endotheliales ICAM-1/CD54 und VCAM-1/CD106 mit Hilfe der Durchflusszytometrie bestimmt. Der Nachweis der vermittelnden monozytären CRH-Rezeptoren R1/-R2 erfolgte mittels RT-PCR- und Immunfluoreszenztechnik. THP-1 konnte als Zielzelle für CRH mit Nachweis der CRH-Rezeptoren auf mRNA- und Proteinebene identifiziert werden. CRH induzierte eine signifikante zeit- und konzentrationsabhängige Adhäsionszunahme der THP-1 Zellen am HMEC-1 Monolayer. Der Effekt scheint Monozyten-vermittelt, da CRH, konzentrationsabhängig, zu einer monozytären MAC-1/CD11b-Freisetzung führte. Eine CRH-Stimulation nur von HMEC-1 führte hingegen zu keiner Adhäsionszunahme, erklärbar z. B. durch die hier dokumentierte fehlende Veränderung von endothelialem ICAM-1/CD54 und VCAM-1/CD106 unter Einfluß von CRH. Die Ergebnisse unterstreichen somit die Relevanz von peripherem CRH auf die Monozytenfunktion und Monozyten/Endothel-Interaktion. Sie können einen Beitrag zur Erklärung eines möglichen Zusammenhangs von chronischem Stress (mit konsekutiver Erhöhung des Stresshormons CRH) und der Initiation / Progression der endothelialen Dysfunktion leisten (Wilbert-Lampen, Straube et al., 2006).

Epidemiologische Daten und Tiermodelle geben Hinweise auf eine verschlechterte Immunabwehr bei Karzinomen des Kopf- und Halsbereiches, wobei die genauen Wirkmechanismen bisher nicht aufgeschlüsselt werden konnten. In Untersuchungen konnte gezeigt werden, dass Karzinome im Kopf- und Halsbereich mit erhöhten Konzentrationen an Prostaglandinen im Serum einhergehen, ein anderer Autor wies eine verminderte Expression des Chemokinrezeptors CCR5 auf Monozyten nach Behandlung mit Prostaglandin nach. In der vorliegenden Arbeit konnte erstmalig aufgezeigt werden, dass es im Rahmen einer Karzinomerkrankung im Hals- und Kopfbereich zu strukturellen und funktionellen Defiziten der Monozytenfunktion kommt. Hierbei finden sich erniedrigte Expressionsraten an CCR5, CCR2 und des Adhäsionsmoleküls CD11b des β-Integrins Mac-1 sowie eine verminderte Adhäsionsfähigkeit der Monozyten an das interzelluläre Adhäsionsmolekül ICAM-1. Weiterhin konnte nachgewiesen werden, dass nach Inkubation der Monozyten in Serum gesunder Spender die verminderte Immunfunktion wieder verbessert wird. Parallel dazu zeigte sich, dass die oben beschriebenen Immundefekte auf Monozyten gesunder Spender durch Inkubation in Serum von Tumorpatienten künstlich herbeigeführt werden können. Dies legt den Schluss nahe, dass im Rahmen einer Karzinomerkrankung Metaboliten im Serum gelöst sind, die eine immunologische Tumorabwehr erschweren. Diese Erkenntnisse sind ein weiterer Schritt zum Verständnis der Tumorimmunologie und könnten dazu hilfreich sein, immunologische Therapieverfahren voranzubringen.

Ziel der vorliegenden Arbeit war es zu zeigen, ob Fibrinogen im reperfundierten Herzen die Interaktion von Thrombozyten und polymorphkernigen Granulozyten (PMN), sowie die Ausbildung von Thrombozyten-PMN Koaggregaten vestärkt. Zudem war von Interesse, ob diese Koaggregate zum Reperfusionsschaden beitragen und inwieweit der GPIIb/IIIa Rezeptor Antagonist Abciximab (c7E3Fab) die PMN-Thrombozyten Interaktion inhibiert und dadurch den myokardialen Reperfusionsschaden vermindert. Die Expression von MAC-1 auf PMN und GPIIb/IIIa auf Thrombozyten wurde mit Hilfe monoklonaler Antikörper gegen CD11b und CD41 im FACS gemessen. Die Versuche erfolgten vor und nach der Koronarpassage durch ein postischämisches isoliertes Meerschweinchenherz, sowie mit und ohne c7E3Fab/LPM19c Inkubation. PMN/Thrombozytenkoaggregate wurden im koronaren Effluat mittels Flusszytometrie und im koronaren Gefäßsystem durch Videofluoreszenzmikroskopie quantitativ und qualitativ untersucht. Die Erholung der externen Herzarbeit wurde an Herzen ohne Zellinfusion, mit Infusion von Thrombozyten und PMN, sowie mit zusätzlicher c7E3Fab beziehungsweise LPM19c Inkubation bestimmt. In der vorliegenden Arbeit konnte gezeigt werden, dass die Reperfusion von ischämischem Myokard die fibrinogenabhängige Interaktion von PMN und Thrombozyten verstärkt. Über die Fibrinogenrezeptoren GPIIb/IIIa auf Thrombozyten und MAC-1 auf PMN kommt es zur Ausbildung von Koaggregaten. Die Analysen der epikardialen Mikrozirkulation mit Hilfe der Doppelfluoreszenzmikroskopie zeigten, dass sowohl homogene Thrombozytenaggregate, als auch heterogene PMN–Thrombozyten Koaggregate im Kapillarsystem reteniert werden. Der durch die PMN–Thrombozyten Interaktion verursachte funktionelle Schaden, konnte in Anwesenheit der Fibrinogenrezeptor Antikörper c7E3Fab (GPIIb/IIIa, Thrombozyten) und LPM19c (MAC-1, PMN) verhindert werden. Die dargestellten Untersuchungen bestätigten die Beteiligung von Thrombozyten am Reperfusionsschaden. Im Mittelpunkt stand dabei die Bindung von GPIIb/IIIa auf Thrombozyten über Fibrinogen als Brückenmolekül an MAC-1 auf PMN. Allerdings wurde die Bildung von Koaggregaten auch in Abwesenheit von Fibrinogen beobachtet, was auf mögliche alternative Interaktionsmechanismen schließen lässt. Weiterhin wurde beobachtet, dass c7E3Fab zwar nicht direkt mit dem für die MAC-1 Detektion verwendeten Antikörper konkurriert, aber die Hochregulation von MAC-1 nach Koronarpassage und die Bindung von Fibrinogen an PMN abschwächt. Eine Erklärung für den Abfall der MAC-1 Detektion nach c7E3Fab Inkubation könnte sein, dass es durch die Blockade der fibrinogenabhängigen Interaktion der beiden Zellkompartimente zu einer verringerten Aktivierung der Leukozyten durch Thrombozyten kommt und damit weniger MAC-1 exprimiert wird. c7E3Fab verhindert die Thrombozytenaggregation, inhibiert die fibrinogenabhängige Interaktion von Thrombozyten und Leukozyten und trägt so möglicherweise zu einer verringerten Aktivierung von MAC-1 auf PMN bei.

Die Differentialdiagnose entzündlicher Hauterkrankungen setzt seitens des Dermatologen ein hohes Maß an klinischer Erfahrung voraus. Labortechnische und histologische Untersuchungsverfahren sind dabei nur eingeschränkt hilfreich. Ausgehend von der Annahme, dass erstens das atopische Ekzem ein krankheitsspezifisches Mikromilieu in der Epidermis und Dermis aufweist und dass sich zweitens dieses spezifische Mikromilieu im Phänotyp der immunkompetenten CD1a positiven epidermalen dendritischen Zellpopulation widerspiegelt, wurde seit 1995 ein Verfahren zur Immunphänotypisierung von Langerhans Zellen und IDEC entwickelt. Es konnte gezeigt werden, dass das atopische Ekzem durch eine hohe Expression des hochaffinen IgE-Rezeptors (FceRI) bei niedriger Expression des IgG-Rezeptors FcgRII auf den dentritischen Zellen der Epidermis hoch sensitiv und spezifisch von anderen Dermatosen abgegrenzt werden kann. Ziel der vorgelegten Arbeit war es, die standardisierte Phänotypisierung CD1a positiver epidermaler Zellen so zu erweitern, dass neben dem atopischen Ekzem auch die Diagnose anderer entzündlicher Hauterkrankungen möglich wird. Es wurden 296 Patienten mittels standardisierter Phänotypisierung epidermaler dendritischer Zellen untersucht. Bei 192 Patienten konnte nach Abschluss aller klinisch diagnostischen Untersuchungen eine eindeutige Diagnose gestellt werden, so dass die an diesen Patienten erhobenen Phänotypisierungsdaten für die Analyse krankheitsspezifischer Veränderungen im Oberflächenmarkerprofil von Langerhans Zellen und IDEC herangezogen wurden. Die Patienten verteilten sich auf 8 Gruppen: Normale Haut (n=31); Atopisches Ekzem (n=59); Psoriasis (n=41); Allergisches Kontaktekzem (n=23); Kontaktekzem bei atopischer Diathese (n=14); Lichen ruber (n=6); Mycosis fungoides (n=10) und Nummuläres Ekzem (n=8). Es konnte gezeigt werden, dass der prozentuale Anteil epidermaler dendritischer Zellen in Abhängigkeit von der Diagnose variiert und dass die bei allen entzündlichen Dermatosen nachweisbare Vermehrung epidermaler dendritischer Zellen vorrangig auf eine de novo Einwanderung von IDEC in die Epidermis zurückzuführen ist. Das wesentliche Ergebnis der vorgelegten Arbeit ist die Erweiterung der diagnostischen Phänotypisierung auf die Psoriasis und das allergische Kontaktekzem. So konnte die Psoriasis durch eine Ratio von CD64/CD11b auf IDEC sinnvoll abgegrenzt werden. Das Kontaktekzem war mittels einer Ratio aus E-Cadherin/CD86 auf Langerhans Zellen zu identifizieren. Die Gruppe der Patienten mit Kontaktekzem bei atopischer Diathese wurde durch die Ratio E-Cadherin/CD80 auf IDEC erkannt. Die bereits für das atopische Ekzem publizierte Ratio FcRI/CD32 auf allen CD1a positiven Zellen wurde mit der gleichen Ratio, ermittelt nur für IDEC, verglichen. Eine Verbesserung der Sensitivität und Spezifität für die Diagnosestellung des atopischen Ekzems konnte hierdurch nicht erreicht werden. Die starke Expression von FceRI auf LC und IDEC kennzeichnet das atopische Ekzem. Typisch für das atopische Ekzem ist weiterhin die Expression von CD36 auf LC und IDEC. Die Psoriasis ist durch eine besonders starke Expression der Fcg-Rezeptoren auf IDEC gekennzeichnet. Darüber hinaus findet sich eine erhöhte passive Anlagerung von sCD23 an IDEC bei Psoriasis. Gegenüber anderen Diagnosen ist die Expression von CD11b und CD11c auf IDEC vermindert. Für das Kontaktekzem ist die im Vergleich mit anderen entzündlichen Dermatosen niedrige CD49d Expression auf LC kennzeichnend. Die Einführung neuer diagnostischer Algorithmen in die Phänotypisierung epidermaler dendritischer Zellen konnte das diagnostische Spektrum dieser Methode deutlich erweitern. Neben atopischem Ekzem und normaler Haut wurden auch Algorithmen für die Psoriasis und das allergische Kontaktekzem beschrieben. Die systematische Analyse weiterer Dermatosen mit epidermaler Beteiligung dürfte auch für diese Erkrankungen diagnostische Algorithmen aufzeigen, die das Mikromilieu der entsprechenden Krankheiten widerspiegeln. Die Phänotypisierung epidermaler dendritischer Zellen dürfte insbesondere im Rahmen klinisch-therapeutischer Studien vermehrt zur Anwendung kommen, da mit dieser Methode eine objektive, quantitative Untersuchung einzelner behandelter oder unbehandelter Hautareale im zeitlichen Verlauf möglich ist.

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 der akuten myeloischen Leukämie (AML) stellen die unkontrollierte Proliferation und Reifungsblockade myeloider Vorläuferzellen, Expansion dieser Zellen in das periphere Blut, extramedulläre Manifestationen und verminderte Elimination der Leukämiezellen durch das Immunsystem grundlegende Pathomechanismen dar. Diese Vorgänge werden über ein komplexes Zusammenspiel von Zytokinen und Adhäsionsmolekülen reguliert. In dieser Arbeit wurde daher mittels Durchflußzytometrie die Expression von Zytokinrezeptoren, Adhäsions- und kostimulatorischen Molekülen in Knochenmarks(KM-) Proben von 103 AML-Patienten bei Diagnosestellung und acht gesunden Probanden untersucht. Zytokinrezeptoren weisen bei der normalen Hämopoese ein reifegradabhängiges und linienspezifisches Expressionsmuster auf. Es wurden daher zum einen Zytokinrezeptoren ausgewählt, die schon in der frühen Hämopoese exprimiert werden, wie der SCF-R (CD117), FL-R (CD135), IL-3-R (CD123) und zum andern Zytokinrezeptoren, die erst in späteren Differenzierungsstadien der monozytären Zelllinie (v.a. GM-CSF-R; CD116) und der granulozytären Zelllinie (v.a. G-CSF-R, CD114) exprimiert werden. Die gp130-Subunit (CD130) stellt die signaltransduzierende Untereinheit von IL-6, IL-11, LIF etc. dar und wirkt synergistisch auf allen Stufen der Hämopoese mit. Die untersuchten Adhäsionsmoleküle wurden in drei Gruppen unterteilt: a) Adhäsionsmoleküle, die den Kontakt zur KM-Matrix oder zu sich selbst beeinflussen: VLA-2 (CD49b), VLA-3 (CD49c) und die erst kürzlich auf hämopoetischen Zellen gefundenen Adhäsionsmoleküle PRR-1 und PRR-2. b) Adhäsionsmoleküle, die den Kontakt zum Endothel fördern: LFA-1 (CD11a), Mac-1 (CD11b), L-Selektin (CD62L) und UPA-R (CD87) c) kostimulatorische Moleküle, die eine Rolle bei der Interaktion der Leukämiezellen mit immunkompetenten Zellen spielen: ICAM-1 (CD54), LFA-3 (CD58), B7-1 (CD80), B7-2 (CD87) und NCAM (CD58). Eine KM-Probe wurde als positiv gewertet, wenn mehr als 20% der Blasten im Auswertefenster den entsprechenden Marker exprimierten. Ergebnisse: Der durchschnittliche Anteil Zytokinrezeptoren exprimierender Zellen war in KM-Proben von AML-Patienten deutlich höher als in KM-Proben von gesunden Probanden. Einzige Ausnahme bildete die gp130-Subunit, die nur auf durchschnittlich 4% der AML-Blasten exprimiert wurde, während durchschnittlich 23% der Zellen in gesunden KM-Proben die gp130-Subunit exprimierten. Bei den Adhäsionsmolekülen zeigte sich im Vergleich zu den gesunden KM-Proben bei der AML ein höherer Anteil von Zellen, die kostimulatorische und Endothel-Kontakt-fördernde Moleküle exprimierten, während der Anteil von Zellen, die das Stroma-Kontakt-fördernde ß1-Integrin VLA-2 exprimierten, vermindert war. VLA-3 konnte dagegen in keinem der untersuchten AML-Fälle und der gesunden KM-Proben als positiv gewertet werden. Innerhalb der AML-Subtypen konnte ein reifegrad– und linienabhängiges (monozytäres, granulozytäres) Verteilungsmuster der Zytokinrezeptoren festgestellt werden: Blasten unreifer Leukämien (M0; M1) exprimierten bevorzugt SCF-R und FL-R. Blasten von AML-Subtypen, die der granulozytären Differenzierungslinie zugeordnet werden (M2, M3), exprimierten v.a. G-CSF-R. Blasten monozytärer Leukämien (M4, M5) exprimierten v.a. GM-CSF-R und FL-R. Der IL-3-R wurde in fast allen AML-KM-Proben auf einem Großteil der Blasten exprimiert. Den größten Anteil positiver Zellen für Adhäsions- und kostimulatorische Moleküle (Integrine, B7-2, NCAM, UPA-R) wiesen die monozytären Leukämien auf. B7-1 wurde v.a. auf Blasten des FAB-Typs M3 exprimiert. L-Selektin, ICAM-1 und PRR-1/PRR-2 zeigten eine variable Expression innerhalb aller FAB-Typen. In der Gruppe der sekundären Leukämien waren signifikant mehr Fälle Mac-1-positiv als in der Gruppe der primären Leukämien (p = 0.074, Qui2-Test). Ansonsten zeigten sich zwischen primären und sekundären Leukämien keine signifikanten Unterschiede. Wichtig für die Entscheidung über Art und Intensität der Therapie bei der AML ist das Abschätzen der Prognose eines Patienten bei Diagnosestellung. Bislang werden Patienten v.a. anhand zytogenetischer Untersuchungen von Karyotypanomalien in Prognosegruppen eingeteilt. Da aber nur ca. 50-60% der AML-Patienten chromosomale Veränderungen aufweisen, besteht ein Bedarf an Karyotyp-unabhängigen Prognosekriterien. Zytogenetische Analysen wurden bei allen AML-KM-Proben durchgeführt und die Expression der Marker sowohl mit den zytogenetischen Risikogruppen als auch mit dem tatsächlichen klinischen Verlauf der Patienten korreliert. In die klinische Auswertung wurden nur Patienten (n = 55) eingeschlossen, die nach dem Therapieprotokoll der German AML-Cooperative-Group behandelt worden waren. In der zytogenetisch günstigen Prognosegruppe zeigten sich im Vergleich zur zytogenetisch ungünstigen Prognosegruppe signifikant mehr G-CSF-R-positive Zellen (p = 0.027, T-Test), signifikant weniger L-Selektin-positive Fälle (p = 0.037, Qui2-Test) und signifikant mehr Mac-1- und PRR-1-positive Fälle (p = 0.005; p = 0.009; Qui2-Test). Diese Marker zeigten aber keine signifikanten Unterschiede bezüglich Remissionrate und progressfreier Überlebenswahrscheinlichkeit der Patienten. Dies läßt sich auf die zum Teil geringe Fallzahl und die kurze Beobachtungsdauer von im Mittel 11 Monaten nach Remission erklären. Andere Marker zeigten dagegen keine Korrelation mit den zytogenetischen Risikogruppen, dagegen aber mit dem tatsächlichen klinischen Verlauf der Patienten: VLA-2-, NCAM-, UPA-R-positive Leukämien zeigten eine signifikant niedrigere Remissionsrate (p = 0.049, p = 0.03, p = 0.03, Qui2-Test). Patienten, in deren KM-Proben >85% der Blasten den FL-R oder >45,5% den SCF-R exprimierten, wiesen eine signifikant niedrigere Wahrscheinlichkeit für progressfreies Überleben auf, ebenso wie Patienten, in deren KM-Proben >60,5% der Blasten ICAM-1-, >15% B7-1-, >65% B7-2- und >8% NCAM-positiv waren. NCAM korrelierte als einziger Marker negativ sowohl mit der Remissionsrate, als auch mit der progressfreien Überlebenswahrscheinlichkeit, allerdings nicht mit der Einteilung in zytogenetische Risikogruppen. Auch für die übrigen Marker konnten Cut-off-Werte für den Anteil Marker-positiver Blasten ermittelt werden, bei denen aus dem Vergleich der entstandenen Gruppen ein deutlicher Unterschied in der Dauer der progressfreien Überlebenszeit hervorging. Diese Unterschiede waren allerdings aufgrund der geringen Fallzahl nicht signifikant, so dass sich eine eindeutige prognostische Aussagen nicht treffen ließ. Dabei wiesen Patienten mit einem höheren Anteil von G-CSF-R-, GM-CSF-R- und einem niedrigeren Anteil von IL-3-R-exprimierenden Blasten eine längere progressfreie Überlebenszeit auf. Patienten mit sehr hohem Anteil PRR-2- oder mit geringem Anteil PRR-1-positiver Blasten tendierten zu einer eher kürzeren progressfreien Überlebenszeit. Umgekehrt wies eine niedrige Expression von Endothel-Kontakt fördernden Oberflächenmolekülen, wie z.B. L-Selektin, Mac-1 und UPA-R auf eine schlechte Prognose hinsichtlich der Dauer des progressfreien Überlebens hin. Therapeutische Konsequenzen: Die in dieser Arbeit aufgezeigten Zusammenhänge zwischen der Expression bestimmter Oberflächenmarker und dem klinischen Verlauf der Patienten helfen, die Prognoseeinschätzung von Patienten - über die Zytogenetik hinaus - weiter zu spezifizieren: So stellt die NCAM-positive Leukämie eine eigene Entität mit prognostisch schlechtem Verlauf unabhängig vom Karyotyp dar. Bei UPA-R- und/oder VLA-2-positiven AML-Fällen sollten aufgrund der verminderten Remissionswahrscheinlichkeit intensivere therapeutische Induktionstherapien eingeleitet werden. Für die Remissionsdauer ist sowohl die hohe Expression kostimulatorischer Moleküle, als auch die hohe Expression von Zytokinrezeptoren, die v.a. auf Stammzellebene wirksam sind und die die Expression von diesen kostimulatorischen Molekülen fördern, prognostisch ungünstig. Diese Patienten sollten bei intensiver Konsolidierungstherapie engmaschig kontrolliert werden und die Indikation zur Knochenmarkstransplantation sollte frühzeitig gestellt weren. In der Zytokintherapie werden G-CSF und GM-CSF regelmäßig in der Klinik zur Verkürzung der Neutropeniephase nach Chemotherapie eingesetzt. Dagegen konnte mit dem Einsatz von G-CSF und GM-CSF als Priming-Medikamente bisher noch kein eindeutiger klinischer Benefit für die Patienten erzielt werden. Die in dieser Arbeit vorgestellten Ergebnisse einer linienspezifischen und reifegradabhängigen Expression der Zytokinrezeptoren legen nahe, dass G-CSF als Primingmedikament v.a. bei granulozytär-differenzierten AML-Subtypen und GM-CSF eher bei monozytär-differenzierten AML-Subtypen eingesetzt werden sollte. In der Supportivtherapie, bei der die Stimulation von AML-Blasten nicht mehr gewünscht ist, sollten G- und GM-CSF genau umgekehrt eingesetzt werden. Da eine hohe Expression von FL-R und SCF-R mit einer schlechten Prognose für die Dauer des progressfreien Überlebens korrelierte, kann sich eine Stimulation dieser Rezeptoren durch die Gabe von SCF und FL in der Supportivtherapie eher ungünstig auswirken, ebenso wie beim Priming, da auch gesunde Stammzellen stimuliert und damit sensibler gegen Zytostatika werden. Darüber hinaus geben diese Ergebnisse auch Hinweise auf mögliche pathobiologische Bedeutungen und damit verbundener neuer therapeutischer Strategien bei der AML: So kann die erhöhte FL-R-Expression - wie bei der Tandemduplikation des FL-R auch - zu einer erhöhten, prognostisch ungünstigen Phophorylierung von Tyrosinkinasen führen. Auch der SCF-R aktiviert intrazellulär Tyrosinkinasen. Neue Medikamente, wie z.B. Tyrosinkinase-Inhibitoren, oder Dexamethason, das die FL-R-Expression auf den AML-Blasten herunterreguliert, könnten bei diesen AML-Patienten neue benefit-bringende therapeutische Möglichkeiten darstellen. Ebenso scheint die Immunantwort bei AML-Patienten trotz, oder vielleicht sogar gerade bei Expression von kostimulatorischen Molekülen vermindert zu sein, was die Gabe von immunstimulierenden Medikamenten, wie rIL-2 oder CTLA-4-Inhibitoren im Bereich der Immuntherapie sinnvoll erscheinen lässt. So leistet diese Arbeit nicht nur einen Beitrag zur Diagnostik, Prognose und Biologie der AML, sondern entwickelt in Zusammenschau mit bereits publizierten Daten neue, therapeutische Möglichkeiten für die Behandlung der AML.

Die entzündlich veränderte Epidermis unterscheidet sich von der Epidermis der normalen Haut durch das Auftreten einer zweiten dendritischen, von Langerhans Zellen (LC) abgrenzbaren Zellpopulation. Während Herkunft und Funktion dieser Inflammatorischen Dendritischen Epidermalen Zellen (IDEC) noch unbekannt sind, deuten die bislang durchgeführten immunphänotypischen Untersuchungen auf eine Monozyten-abhängige Genese hin. In der vorgelegten Arbeit wurden Untersuchungen an verschiedenen epidermalen Zellsuspensionen, sowie in vitro aus Monozyten hergestellten unreifen dendritischen Zellen (MoDC) durchgeführt, um durch eine standardisierte Untersuchung weiterführende Erkenntnisse zur Immunbiologie dieser dendritischen Zellen zu gewinnen. Zunächst konnte gezeigt werden, daß die in vitro hergestellten MoDC immunphänotypisch relativ genau den IDEC entsprechen, während der Immunphänotyp der LC deutliche Unterschiede zu dem der MoDC aufweist: Während sowohl IDEC als auch MoDC stark CD36, CD11b und CLA exprimieren, sind auf LC CD36 und CD11b nur minimal und CLA deutlich weniger stark exprimiert. Außerdem konnte gezeigt werden, daß die CD1a- Expression der MoDC mit längerer Kulturdauer stärker wird, und daß eine Zugabe von autologem Serum in die Kultur diese Ausreifung von Monozyten zu MoDC hemmt. Der zweite Teil dieser Arbeit befasste sich mit vergleichenden Zellzyklusuntersuchungen von LC und Keratinozyten aus normaler und entzündlich veränderter Haut. Hierzu wurde zunächst eine Untersuchungsmethode zur gleichzeitigen Bestimmung von Zellzyklus, CD1a- Expression und Vitalität etabliert. Es wurden verschiedene Parameter wie Zellzahl, Farbstoffkonzentration, Fixierung und Vitalfärbung an einem auf den Zellinien MOLT4 und U937 basierenden Modell optimiert, und später auf epidermale Zellsuspensionen übertragen. Dieses aus einem Zellgemisch einer CD1a-exprimierenden und einer CD1a-negativen Zellinie in seinen Anteilen frei wählbare Modell erlaubte die Optimierung der Färbetechnik und wies den vermehrten DNA-Gehalt der MOLT4-Zellen nach, was auf eine chromosomale Aberration in der MOLT4-Zellinie hindeutet. Keratinozyten und Langerhans Zellen wiesen einen normalen Zellzyklus innerhalb der normalen Haut auf, was die Resultate anderer Arbeitsgruppen bestätigte. In entzündlicher Haut war die Zellteilungsrate bei Keratinozyten und epidermalen dendritischen Zellen erhöht. Die erhöhte Zellteilungsrate der epidermalen dendritischen Zellen war bislang noch nicht untersucht worden. Schließlich wurden funktionelle Untersuchungen der immunphänotypisch nachgewiesenen kostimulatorischen Moleküle durchgeführt. Hierzu wurde die antigenpräsentierende Funktion mit der gemischten Haut-Lymphozyten-Reaktion untersucht. Durch den CD86 Antikörper konnte eine T-Zell-Proliferation in vitro effektiv gehemmt werden. Zusammenfassend konnte in dieser Arbeit eine Methode zur Untersuchung epidermaler dendritischer Zellen etabliert werden. IDEC ähneln in zahlreichen Aspekten den MoDC, wogegen LC deutliche Unterschiede aufweisen. Weitere Untersuchungen am Zellzyklus entzündlicher Haut, die auf die in der vorgelegten Arbeit etablierten Methode aufbauen, werden Aufschluß über die Teilungsraten von LC und IDEC geben. Anhand der Erkenntnisse über die Rezeptorexpression auf IDEC und MoDC ist eine genauere immunbiologische Einordnung der IDEC in die Gruppe der myeloiden dendritischen Zellen wahrscheinlich.

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.04.22.055228v1?rss=1 Authors: Hollinger, K. R., Sharma, A., Tallon, C., Lovell, L., Thomas, A. G., Zhu, X., Kambhampati, S. P., Liaw, K., Sharma, R., Rojas, C., Rais, R., Kannan, S., Kannan, R. M., Slusher, B. Abstract: Roughly half of all individuals with multiple sclerosis (MS) experience cognitive impairment, but there are no approved treatments that target this aspect of the disease. Recent studies link reduced brain N-acetylaspartylglutamate (NAAG) levels to impaired cognition in various neurological diseases, including MS. NAAG levels are regulated by glutamate carboxypeptidase II (GCPII), which hydrolyzes the neuropeptide to N-acetyl-aspartate (NAA) and glutamate. Although several GCPII inhibitors, such as 2-(phosphonomethyl)-pentanedioic acid (2-PMPA), elevate brain NAAG levels and restore cognitive function in preclinical studies when given at high systemic doses or via direct brain injection, no GCPII inhibitors are clinically available due to poor bioavailability and limited brain penetration. Systemic hydroxyl dendrimers (~4 nm) have been successfully used to enhance brain delivery of drugs selectively to activated glia. We recently discovered that GCPII is highly upregulated in activated microglia after brain injury. To determine if dendrimer conjugation could enhance the brain delivery of GCPII inhibitors, specifically in the context of MS, we attached 2-PMPA to hydroxyl polyamidoamicne (PAMAM) dendrimers (D-2PMPA) using a highly efficient click chemistry approach. Targeted uptake of D-2PMPA into activated glia was subsequently confirmed in glial cultures where it showed robust anti-inflammatory activity, including an elevation in TGF and a reduction in TNF. Given these positive effects, D-2PMPA (20mg/kg) or vehicle dendrimer were dosed twice weekly to experimental autoimmune encephalomyelitis (EAE)-immunized mice starting at disease onset (therapeutic paradigm). D-2PMPA significantly improved cognition in EAE as assessed by Barnes maze performance, even though physical severity was not impacted. Glial target engagement was confirmed, as CD11b+ enriched cells isolated from hippocampi in D-2PMPA-treated mice exhibited almost complete loss of GCPII activity. These data demonstrate the utility of hydroxyl dendrimers to enhance brain penetration and support the development of D-2PMPA to treat cognitive impairment in MS. Copy rights belong to original authors. Visit the link for more info