Autoimmunity - Science topic

CAR T quashes autoimmune diseases

"Three people with severe autoimmune diseases are in remission after being treated with bioengineered and CRISPR-modified immune cells called chimeric antigen receptor (CAR) T cells. They are the first to be treated with engineered immune cells from donors. The advance could represent the first step towards mass production of CAR T therapies for conditions such as lupus and multiple sclerosis. The ongoing success and safety of the therapy need to be demonstrated in more people before it can be considered for wider use, but it “could prove paradigm shifting”, says immunologist Daniel Baker..."

Hi Zarmina,

The list of articles and answer by Malcolm Nobre are quite comprehensive. I would like to suggest these two reviews on OLP:

The human microbiome in immunobullous disorders and lichen planus.

The immunopathogenesis of oral lichen planus-Is there a role for mucosal associated invariant T cells?

Kind regards

Yakubu Lawal

Well; the deduction (in the former replies) that your concern is (by epidemiology) to be near Kano, Katsina or even Kaduna, is precisely the case. That’s quite a pinpoint deduction considering the vast size and variety of Nigeria (we hope you agree). Would you please consider mentionning if you are studying a generality (for Nigeria) or a pseudo-cluster (epidemic for Bauchi) or an individual case? It matters not if it’s something “rare” or only one patient. We’re very interested in helping if you may share it.

Europe is saturated in glycoalkaloid and Rheumatoid Arthritis while in Africa you may have the best opportunity to detect the very start of any (or all) of the NCDs that are “new” or never seen before.

My work is based on the facts of potato and eggplant glycoalkaloids having been underestimated (in Europe). South America gave very serious warnings (about the toxic tubers and their subtle idiosyncratic poisonings; effectively identifying them as a failed project of domestication from their point of view). Denmark heeded the warnings precisely because the originators lacked any ulterior motive to praise or promote the tubers nor even to pass them off as inoffensive, rather firm that they were/are dangerous. In the extreme, in Ukraine, some use the potato to suicide. The toxic tubers were banned for human consumption in Denmark until the pressure of war in 1914. There’s nothing traditional about it in Europe (although there is marketing inertia). There’s nothing superstitious about it either (although some pressure keeps leaking from Europe into the United Nations to shut down any objective examination of the tuber’s dubious history).

Nigeria. GEZ (Germany) are recommending another push since 2021, called the “Potato Revolution for Bauchi”. It’s a common feature (in Africa) with only one advice, to “just cut off the green bits”. This is simply not good enough. The German Food and Safety office did note (to the EFSA) that modern cross-breeding of potatoes suppresses any and all green tints. Advice from the 1920s was (according to ITT research) false in the first place and now irrelevant. The point is that where farmers attempt potato and then industrialists learn (from their efforts) that the tuber will grow at all, there’s always pressure to attempt to profit from it (for export in the case of East Africa).

Admittedly this response may seem terse yet in favour of some new medical point of view concerning human health. Much more likely to occur in Africa because the arrival of glycoalkaloids to market is “new” or within a few recent generations in certain zones.

If your examination concerns something seen since a long time with historical confirmation, then glycoalkaloids may merit a sharp look because they’re also in these particular plants too:

Solanum Nigrum (glycoalkaloid Solasodine; boiled to extract/remove)

Solanium Marcrocarpon, Gboma, Akwukwo Anara (Igbo), Efo Igbo (Yoruba), Ganyen Gauta (Hausa) or “Garden Egg Leaf”.

For a full list (for Azare), Albashir Tahir (Bauchi State University) knows the topic and the experts.

If your examination concerns something recent and unexpected then Aubergines/Eggplants or Potato promotion may be a much more likely “new” local contemporary cause.

Note concerning your original question: The medical colleagues who’ve noticed the indications of Latitude and Sunshine also measure a conflict in the data concerning all NCDs, particularly in Scandinavia. In the meantime we hope to advise that they’re actually noticing the zones where potatoes are planted, either because of abundant water, irrigation, suitable soil and potato growing or where nothing else will, then consumed with high Fatty Acid foods, so causing the highest chemical pathogen rates. Although usually in colder climes, that’s no longer the case when a short season well watered is “targetted” (to use the Finland Crop Research project terms about Tanzania).

Here, our future report (partly endocrinology) will cover the subject and all at proof level. However, those such as in the team of Professor Juraj Sepčić (also on Researchgate) preferred to study the epidemic or cluster (in Croatia) using the standard Odds Ratio method to catch the cause as soon as possible. Then it’s objective, in the field of epidemic and free of opinion or delay.

Please let me know your thoughts. By the way I’m a signal processing Engineer (studying metabolism and epidemiological data furnished by the University of Washington compared with basic nutritional supplies data from the United Nations FAO). It's support expertise, without a doctorate in medicine. Yours sincerely, Matthew.

Hi Esther Nutifafa Senanu

My suggestion would be to work along the lines of a paper from Kragstrup and colleagues published in 2019.

Even though mesenchymal synoviocytes display a lot of properties of epithelial cells in monolayer/ 2D cultures (while lacking desmosomes/ tight junctions). To that end I would concentrate on a cell culture model for psoriasis for which you have culture models available that are summarized here:

You can even set up models for ultra-rare diseases like epidermolysis bullosa, which while not autoimmune diseases in the classical sense, they have a strong inflammatory component, in more detail described here:

In general, broad targeted analysis of the proteome (like in the Kragstrup paper) should help with the analysis of these (depending on your experimental choice) increasingly complex models.

I hope that helps!

Good luck experimenting,

Michael

Hello Aradhana Rani

You may refer to the article attached below. This review describes the toxicities caused by CAR T-cells and reviews the published approaches used to manage toxicities.

Toxicities caused by CAR T-cells are diverse and not fully understood. Management requires vigilant monitoring, aggressive supportive treatments, and, in some cases, intensive care. Administering immunosuppressive agents to decrease toxicity is an evolving practice. Overall improvement in the field of CAR T-cell therapies could be achieved by improving the management of CAR T-cell toxicity.

Best.

Studies clearly suggest there is a difference in miRNA expression in plasma & serum. Begin from sample collection, RNA isolation, cDNA synthesis, qPCR, spikes, and endogenous control influences the process. Furthermore, the sample's condition like normal and disease all matters. So until you do it will be very unpredictable.

Please find some cardio papers that discussed the difference between plasma and serum samples for miRNA expression.

Regards

Saurabh

Thank you Muhammad Ghafoor Ali

Are you looking at long COVID-19 infection or post COVID-19 symptoms and infections ?

It has a definite relation

I can give you literature in abundance on this

Thanks a lot!!!!!

I think it is possible

COVID-19 is associated with hyperactive immune responses in both chronically ill and healthy individuals. While autoimmune diseases do experience altered immune system function, even though the ability to fight off pathogens is not necessarily weakened; rather the response is misguided.

Dear Vice-Rector of Research Dr. Normakhamatov!

You spotted a highly relevant point. Humans might slowly toward the future able to develop better immune - resistance with healthier lifestyles, but the more rampage COVID-19 becomes and the more dynamically it can spread out all over the globe the less likely this immune - resistance would develop due to new emerging mutations and climate change:

1) Wagner Gouvêa dos Santos (2021). Impact of virus genetic variability and host immunity for the success of COVID-19 vaccines, Biomedicine & Pharmacotherapy

Volume 136, April 2021, Citation: "Even if COVID-19 may be brought under control in the near future, unexpected outbreaks and development of SARS-CoV-2 resistant to treatments or even vaccines are highly possible due to new mutations."Open access:

2) Gorji, S., Gorji, A. COVID-19 pandemic: the possible influence of the long-term ignorance about climate change. Environ Sci Pollut Res (2021). https://doi.org/10.1007/s11356-020-12167-z Citation: "Multiple lines of evidence suggest that the large-scale and rapid environmental changes in the last few decades may be implicated in the emergence of COVID-19 pandemic by increasing the potential risk of the occurrence and the spread of zoonotic diseases, worsening food security, and weakening the human immune system. As we are facing progressive climatic change, a failure to act accordingly could inevitably lead to further, more frequent confrontations with newly emerging diseases.Open access:

Yours sincerely, Bulcsu Szekely

no, it is not possible. microglia does not circulate outside the brain, so you need the brain to extract it. But: you can derive microglia from murine iPSC lines, modified with your virus, and then inject it into mice in which you have ablated most microglia using a CSF1R inhibitor. Cool experiment this would be....

Covid_19mRNA vaccine is novel one,so great care should be cosidered with autoimmune phenomena because vaccine may trigger the disease

In India, RML Mehrotra Pathology Pvt. Ltd & Dr Lal Path Labs, I guess !!!

Rich countries have more resources to get a quick recovery in general....

That is interesting regarding the poor Treg function in RAG1ko‘s. Sorry that the paper was of no help to you, as I said it was a quick google search but is obviously 2013! Would you be able to let me know what you find out?

Have a look at this useful RG link.

Please also see the following RG links.

High consumption of foods containing tannins, thiols, phenols, misothiocyanates and phycocyanin should be avoided. A list of foods containing these substances includes garlic, leek, chives, onion, mustard (thiols), black pepper, red chilies, mango, pistachio, cashews, aspartame, food additives (phenols), mango, cassava, yucca, guarana, betel nuts, raspberry, cranberry, blackberry, avocado, peach, ginger, ginseng, tea, red wine, coffee, spices, eggplant (tannins), mustard, horseradish, cauliflower (isothiocyanates) and Spirulina platensis alga (phycocyanins).

Neopterin is not assessed commonly in POTCs. You can quantify it only in cental labs or research labs

Please see the below given RG links.

@ Olivier Neyrolles

Please go through the following RG link.

Thanks!

I think immediate cesation of steroids is possible in cases of less than 2 to 3 weeks of therapy.

Dear it may not be the best treatment because you have to look at the cause of the HDN is it ABO incompatibility or Rhesus blood group associated HDN which you will solve in that direction.

Subgroup analysis is feasible, provided that analysis according to the factors that you want to subgroup is provided in included or some of the included papers. Otherwise you should opt for individual patient data analysis. Rather than subrgrouping you can also consider meta-regression analysis taking in mind the factors that you want to adjust for. This might be more feasible in certain cases.

This is a fascinating question. Immune privilege can be thought of as antigens hidden away from the immune system by either physical (i.e. blood-brain barrier) or immunological barriers. Immunological barriers include low expression of MHC proteins, high balance between regulatory and effector cells or expression of protective molecules such as Fas or immune suppressive cytokines such as TGF-beta (as suggested as mechanisms of immune privilege in the anterior chamber of the eye). The reason that females are more susceptible to males for most classical autoimmune diseases, with exceptions being type 1 diabetes, myasthenia gravis, pemphigus vulgaris and Goodpasture's syndrome, is not clear. Some authors have suggested that women make stronger inflammatory responses to infectious agents than men because their survival is more important for continuance of the species. However, the detailed mechanism of this is not clear.

in addition to the very valuable papers from Thomas, take into account of you wanna use the recombinant HNAs or extracted HNAs or the whole cells.

Hello,

Have you tried with Google Scholar? You usually have different websites referenced for one paper, and on some you have access to the full article.

You can also contact the corresponding author (or the first) and ask for the full text.

Thank you for your insightful answers. What I want to understand is: why isn't PBC treated as an autoimmune disease despite its autoimmune etiology?

I do know that it's treated with UDCA and that's the most effective treatment we have so far, but I want to know why.

Still needs more trials though. But I think it's a good start to find molecular evidence behind this;)

agree with @ M. Jameel

regards

I would make SDS/reducing lysates of whatever tissue you think is the target of these antibodies, and analyze by Western blot with patient's serum. Comparing more than one patient's serum as well as normal controls will help you identify specific targets. Remember that human sera will bind a lot of proteins, so be sure to titer it down. After you have identified a protein of specific mass, you can cut it out and do mass spec for identification.

Thank you all for your answers. However, my curiosity is also in line with Geraldo's question. Cancer cells thrive in an immune microenvironment bypassing the body's natural self-defence. A number of pathways have been validated as well in lieu of cancer-immunity crosstalk. Are there reported interactions among transformed cells and immune cells which may alter the morphology of immune cells, keeping in view structural alterations of participating proteins presented on the surface of either category of cells?

Hi Dr. Al-Salhie, unfortunately your link shows an equipment that is called "Crocodile" ( the commercial name of the instrument is crocodile, a device to pipette, wash and incubate plates only). However, Berthold technologies does not sell ELISA assays directed to crocodilian proteins, just as nobody else does for reptiles. Believe me, I searched everywhere for antibodies against snake proteins.

Thank you though, for a split second my hopes were high...

A single amino acid in the milk protein beta-casein makes the difference between A1 and A2. The A2 casein at position 67 contains the amino acid proline, and the A1 casein histidine. Thus, in the digestion of A1, the so-called opiate beta casomorphin-7 (BCM-7) is split off, not in the A2 varinates. This metabolite is said to be the reason that some people can not tolerate milk.

Probable there is a reverse relationship between the climate and the onset of autoimmune diseases. They predominate in humid and cold climates generally speaking

Yes. both your answers were very helpful. Just what i was looking for.

Tank you very much.

Different strategies including the use of depleting/blocking antibodies (CD25, CTLA-4, CCL2), IL-2/immunotoxin conjugates, cyclophosphamide, or COX2 inhibitors have been tested in mouse models of cancer and pre-clinical trials, but these did not always result in decreased numbers of Tregs or good anti-tumour response (see: Beyer & Schultze. 2006. Blood 108(3)804-11. https://www.ncbi.nlm.nih.gov/pubmed/16861339).

Frederick Arce, in Sergio Quezada’s group (University College London Cancer Institute), has recently published a paper in which they report on the importance of the anti-CD25-depleting antibody isotype in mouse models of transplantable tumour cell lines. They found that using engineered anti-CD25 antibodies with increased binding to Fc receptors results in enhanced depletion of intra-tumoral Tregs. This antibody, used in combination with immune check-point therapy led to complete tumour rejection in mice. Their observation that CD25 expression is restricted to CD4+Foxp3+ in human peripheral and tumour-infiltrating lymphocytes led them to propose that this validates the use of CD25 as a target for therapeutic Treg depletion in humans.

Very interesting paper (https://www.ncbi.nlm.nih.gov/pubmed/28410988) to read and see where the field of combined immunotherapy for cancer is moving. Their results suggest that efficient depletion of Tregs would be possible in clinical trials in order to improve the response to other therapies.

You "applaud" to your paper?

Interesting.

Protein arrays might also be an option

Thanks a lot, Michael !

I am using Trizol reagent for RNA isolation and i have isolated RNA easily there was no such issue of erythrocytes or thrombocytes. The thing I am worried about is that most of the defensin's transcripts have been isolated from other tissues not blood now I just hope to get altleast some of the transcripts from blood as well.

There is some older literature supporting and pushing the idea that anti-DNA antibodies are actually anti-protein antibodies that cross-react with DNA, but there is no fundamental reason to adopt this notion.  As Jing Zhang pointed out, antibodies can potentially react with just about any chemical/biochemical molecular target, based purely on serendipitous affinity of non-covalent chemical bonds between the antigen and “complementarity-determining regions” of the antibody.  (In some cases monoclonal antibodies can even react with disparate antigens not because of true cross-reactivity but probably due to alternative or overlapping binding sites within the CDR.) Nevertheless, the process for production of anti-DNA antibodies in SLE is likely the same as that for induction of any antibodies – the presence in the human of a B cell that happens to have BCRs with an above-threshold affinity for native DNA and which receives “help” for its activation and affinity maturation by T cells. The existence of such B cells is explainable by the huge B cell repertoire and the inefficiency of B cell tolerance mechanisms.   The pathological event, in my opinion, requires the co-presence of autoreactive T cells that had not been tolerized to a peptide ligand derived from a protein that is physically associated with DNA.  Histone-reactive T cells would be obvious candidates, and such T cells could also provide help for autoreactive B cells with receptors for histones or for histone-DNA complexes (nucleosomes/chromatin), which are also common in SLE.  In addition naïve autoreactive T cells would probably also require pre-activation by “professional antigen-presenting cells” presenting the same ligand, but how this happens is more of a mystery. You ask why would this happen: the only (unsatisfying) answer is that the immune system is inherently flawed or imperfect.  Autoimmunity is a side-effect of the evolutionary drive to create an immune system that can anticipate and respond to just about any pathogen.

Nice

In breast cancer cells, calcitriol has an antiproliferative effect, but it does not induce apoptosis. Nevertheless calcitriol enhances the apoptotic effect of other antitumoral drugs. You can review the following article Journal of Steroid Biochemistry & Molecular Biology 148 (2015) 122–131

Type 1.5 diabetes is a non-official term that is sometimes used to refer to a form of type 1 diabetes known as Latent Autoimmune Diabetes in Adults (LADA). The term type 1.5 refers to the fact that the condition is a form of type 1 diabetes that can share some features that are more commonly associated with type 2 diabetes.

Type 1.5 diabetes is sometimes also called as type 3 diabetes or “double diabetes,” because the adult has aspects of both Type 1 and Type 2 diabetes. Type 1.5 diabetics are said to have “double” diabetes because they show both the autoimmune destruction of beta cells of Type 1 diabetes and the insulin resistance characteristic of Type 2 diabetes. People with Type 1.5 have autoantibodies and gradually lose their insulin-producing capability, requiring insulin within 5–10 years of diagnosis. As their insulin resistance suggests, many people with Type 1.5 diabetes are obese or overweight.

Misdiagnosis as having type 2 diabetes is common. Around 15-20% of people diagnosed with type 2 diabetes may actually have Type 1.5 diabetes.

Autoimmune reactions often result from infection with a virus. The HLA system cuts pieces (8-10 a.a. for Class I to CD8+, longer for Class II to CD4+ T cells), of viral peptides. This helps in recognition and destruction of infected cells, but if the HLA type captures the same fragment of a normal protein like MBP, this results in auto-immunity. So, the only requirement is for a systemic infection, but viruses which target long-lived structural cells will often trigger more auto-immune reactions than those infecting epithelial or blood cells. So, its more of an HLA and self-T cell repertoire. 

Collagenase D prepared from Clos. histolyticum ihas high collagenase activityand very low tryptic activity. Collagenase 2 has a highLipolytic activity from fat cells

I have tried transfection of cellular RNA into 293T cells by lipofectamine, the cellular RNA did not stimulate the cells to produce IFN. the transfection of RNA by lipofectamine may directly deliver the RNA into cytoplasm that avoid to be recognized by TLRs. 

Dear Monica,

I usually take 200 µl of mouse blood for standard genotyping. How much DNA do you need for your analysis?

if one of the causes or the causes of ME/CFS is chemical exposure, than how autoimmunity may accure leaves the possibility of molecular mimicry open if there is any truth to it, or weither it depends strictly on immune malfunction, loss of homeostasis, than what would fallow would be seceptability to infection, just because viruses are often found does not mean they are the cause.  I know for a fact that severe exposure in a high moisture water damaged building can lead to ME/CFS, I know that I'm highly sensitive to certain chemicals and voc's along with allergens. if molecular mimicry can happen this type of exposure would be a very likely contender, just keeping a open mind and asking questions, but ya, I would think you could see my interest in the subject, I understand your view.

Dear Amit~Chronic Disease is an intracellular infection: www.mpkb.org

Benicar(olmesartan) 40 mg every 4-6 hours, acting as a VDR agonist, will partner with the innate immune system to reverse the autoimmune condition.  The drug therapy can be introduced clinically to patients who are motivated to follow the Marshall Protocol and able to tolerate the immunopathology that results from pathogen die-off.

See www.np-privatepracticeassoc.com for some case study series results over 4 years.

Then register for www.MarshallProtocol.com to check out the worldwide cohort utilizing this method.

Our newest paper:  https://t.co/FY4xMjzFF9 

A prior paper: bit.ly/2af83oU

Please let me know if I can be of further assistance.

Sincerely,

Trudy J. Rumann Heil MS, ARNP, CLNC

Scottsdale, AZ USA

Hi

look PDF attached and these links PLZ

ERAP1 endoplasmic reticulum aminopeptidase 1 [Homo sapiens ...

www.ncbi.nlm.nih.gov/gene?Db...National Center for Biotechnology Information

Jul 3, 2016 - epistatic interaction between ERAP1 and the HLA class I alleles .... have orthologs with human gene ERAP1; Map Viewer (Mouse, Rat) ...

Erap1 MGI Mouse Gene Detail - MGI:1933403 - endoplasmic ...

Mouse Genome Informatics

View mouse Erap1 Chr13:74639872-74691875 with: phenotypes, sequences, polymorphisms, proteins, references, function, ... Erap1. Name. endoplasmic reticulum aminopeptidase 1. Synonyms. Arts1, ERAAP, PILSAP .... Primer pair 1.

QIAGEN - Products for Erap1 (Mouse)

https://www.qiagen.com/shop/genes-and-pathways/gene-details/?geneid...

Qiagen

GeneGlobe Products for Erap1 (Mouse) Full Name: endoplasmic reticulum ... EpiTect Methyl II PCR Primer Assay for Mouse Erap1 (CpG Island 103126).

The Role of Endoplasmic Reticulum-Associated Aminopeptidase 1 in ...

Journal of Immunology

by E Firat - ‎2007 - ‎Cited by 87 - ‎Related articles

We generated ERAP1-deficient mice and analyzed cytotoxic responses upon infection .... the Expand long template PCR system (Roche) with the forward primer.

You've visited this page 2 times. Last visit: 7/28/16

You could use the Coombe's technique (like the test used to diagnose autoimmune haemolytic anaemia)- wash the cells in PBS three times and incubate them with anti human immunoglobulin (antibody against human immunoglobulin which you can buy) and examine for cell clumping. If the cells have antibody on their surface, they will bind to the AHG and clump together. You can visualise the clumps under microscope. This is an easy test to do. Good luck 

Many toxic chemicals have been linked to development of autoimmune thyroid disease (and it is prevalent in women with endometriosis).  The list of chemicals is very long --I'm sure you'll find them in the work of Thomas Zoeller.

Grazie mille Angelo. Speriamo in questo Jak-treatment di cui mi parlavi proprio tu tempo fa.

Kind regards for the articles you forwarded. I will take a look at them this week.

Dear colleague,

I personally have seen changes in bone marrow morphology in lymphoma patients prior to any chemo. Mostly, I saw changes in eritroblasts such as binuclear cell morphology, or mitotic figures. Sometimes I saw micromegakaryocytes or pseudo-Gaucher cells. This is rare events. The origin of this changes is unknown to me. My own speculations may be that malignant cells and their microenvironment may produce some cytokines, or substances that disturb regular hematopoiesis. Of course, this need to be evaluated through serious investigations. 

ERVs have been around for a very long time and have altered immune responses in many species including in humans (see the link). Depending upon the type of ERVs, their numbers in the human genome, and the regulatory sequence they harbor, it is likely that these ERVs in humans contribute to the number of outcomes, including those you mentioned.

Link: http://www.ncbi.nlm.nih.gov/pubmed/?term=endogenous+retrovirus%2C+aim2

Biribidobiribbu'  

Several aspects have to be kept in mind when discussing the persistence of auto-antibodies in the CSF. In the case of paraneoplastic syndromes the source of autoantibodies are presumably plasma cells in secondary or tertiary lymphoid structures outside the CNS. Therefore, levels of antibodies in the CSF should be linked to serum levels. It has to be noted that paraneoplastic antibodies can persist for years, even when the tumor has been removed (Alexopoulos H  et al. Paraneoplastic anti-NMDAR encephalitis: long term follow-up reveals persistent serum antibodies. J Neurol. 2011 Aug;258(8):1568-70).

Dear Anthony,

With all due respect, while it is true that researchers and clinicians choose to measure the TTg antibody response as a convenience, and claim that this is a valid autoimmune response (rather than a result of a collateral reaction associated with a primary reaction against an exogenous antigen), this in no way proves or even infers that the primary inflammatory reaction is not directed at the gliadin and glutelin peptides resulting from the digestion of the gluten found in wheat and related species.

To support that observation I submit the fact that when the exogenous antigen is withdrawn from the diet, the TTg reaction ceases.  Obviously it is not an independent reaction.  So how could the TTg antibody response actually be a valid autoimmune response?  The answer is, "It cannot be a true AI response".  If the reaction were truly autoimmune, then it would continue, independent of the status (presence or absence) of any exogenous antigens in the diet.

Dear Jonathan,

I appreciate your analysis and your clarification.  I agree, it could be misleading to generalize in science. It is also important to accept that biomedical sciences are in search of concrete models that would allow to clarify the cause-and-effect of an observation.  Let’s take the hypothesis that we have to represent at one point of a circle autoimmunity. What would be the antipodal point?  Autoimmunity is cause-and-effect sequence and should exist a model to describe it by establishing: a) a concrete and safe cure for the case of diseases and b) a logic conception so to understand this cause-and-effect phenomenon.  

I think you agree that either in B or T cells, autoimmunity engages a selection system. You nicely said that B cell regulation is based on random diversification and selection rather than instruction.  I could say that instruction is also a selection system.

“Roitt and Doniach at University College London and others have shown that in terms of antibody-producing B lymphocytes, diseases such as rheumatoid arthritis and thyrotoxicosis are associated with loss of immunological tolerance which is the ability of an individual to ignore "self", while reacting to "non-self". “This breakage leads to the immune system's mounting an effective and specific immune response against self-determinants”.  This breakage might lead to autoimmunity.

In the clonal deletion theory, proposed by Burnet and Medawar, self-reactive lymphoid cells are destroyed during the development of the immune system in an individual (Nobel Price 1960 acquired immunological tolerance).

So, both Roit and Burnet and others introduced in their approach of studding autoimmunity the term of “loss or destroy”.  Even if autoimmunity is driven by an ‘instructive pathway”; at the bottom line a healthy immune system has the capacity to make a decision using its “proper internal control” and decides to keep the self-intact and attack the non-self.  . If, autoimmunity engages a false elimination or destruction or an “instructive process” of a target (cell - molecule) which is involved in the autoimmune response then we have to ask what the machinery for this process is.

I could say this includes proteolytic machinery, proteasomes, ubiquitination, apoptotic machinery.  This should be the antipodal point of autoimmunity in the circle and this is illustrated best by the term apoptosis.

Last but not least, talking about AIRE, it should not be a special case and I would not considered it as a monogenic disease, as it is in general claimed. AIRE defines a spatial nuclear domain with concrete properties. So any reference to AIRE makes reference to nuclear space. In the same category, we could add PML, SP100 in PBC and more. Vassilis

I hope that following information will be helpful.

Hi Omer, 

The IC marker HELIOS would be the most conventional way to identify nTreg and this has recently been shown to enhanced by the addition of Nrp1. This has been reviewed by Lin et al. 

Keep in mind autoimmune liver diseases may have high titers of anti-albumin IgG. A sensitve assay can detect them, Is your solid matrix coated with hu or Bovine albumin/

is your buffer contain albumin? Check all and revalidate.

One idea is to isolate sera from  your mice and see if they have antibodies to mitochondria.  I would recommend the Kallestad Hep-2 kit from Biorad.  This is designed to test for anti-nuclear antibodies, but the manual describes other autoantibodies you might observe including those to organelles.  Anti-nuclear antibodies might also arise as a secondary effect in your mice. Just substitute a fluorescent anti-mouse Ig secondary antibody for the one in the kit.  The only caution is that you may need a large cohort of both experimental and control mice (probably at least 20 of each) for statistical significance because autoantibodies do show up in control mice.

 Dear Ghada Abdallah,

There are 2 components to your question. Let me tackle the easy part first. 

1. Evidence for efficacy for Hydroxychloroquin in ANA positive ITP: It was always known that almost 5% of SLE might start with isolated thrombocytopenia alone. Mechanisms of immune thrombocytopenia in SLE differs from those observed in primary ITP and is probably multifactorial, including antiplatelet glycoprotein antibodies such as in ITP but also immune complexes, antiphospholipid antibodies, autoantibodies to the c-MpL receptor and megakaryocytes.

Since Hydroxychloroquin works so well in treatment of multiple domains in SLE, it was but natural to study it in SLE associated throbocytopenia too where it was found useful. Next, about 20% of all ITP patients are ANA positive without meeting the criteria of SLE and this was the natural target to be studied for the effect of hydroxychloroquin. Interestingly, ANA positivity in ITP has been shown to a risk factor of chronic ITP and resistance to classical second-line treatment for ITP.

In the 2014 study by Khellaf et al, they looked at both SLE with thrombocytopenia and ANA positive ITP who had failed multiple drugs and tried hydroxychloroquin on them. As expected the SLE group indeed did very well with 83% response but even the other group showed a robust 50% response. Considering the excellent safety profile of hydroxychloroquin, these numbers are very encouraging. If these findings are replicated, hydroxychloroquin will very likely make its way into ITP treatment guidelines soon.

2. Why does it not work in ANA negative ITP? 

Where is the evidence that hydroxychlororquin "does not" act in ANA negative ITP as stated by you. Evidence is just emerging that it might be useful to some extent in the ANA positive variety. It requires another study to prove its inefficacy" in ANA negative ITP.

It was a natural step to use a time tested drug like hydroxychloroquin with proven efficacy in SLE to the subgroup of ANA positive ITP (who do not fulfil criteria for SLE). The next logical step seems to be to compare role of hydroxychloroquin in ANA positive Vs ANA negative ITP. I am not aware of any study that has specifically looked at this aspect. Maybe you can plan one such study.

If we are able to show in future that hydroxychloroquin "DOES NOT" act in ANA negative ITP, then we need new studies to study molecular mechanisms to understand the "why". 

hope this helps

Mo,

A better and easier alternative for you will be to contact the authors of publication and request them to provide the cells you intend to use.  With a material transfer agreement, most scientists and institutions are willing to share such resources.

If you must create your own cell line, you will have to use identical conditions as described in the original work. And even then, the cells line you isolate and propagate, may or may not be nearly the same, much less identical - another reason to acquire the same strain.

Best...

PARP-1 is involved in to regulate the pathogenesis of inflammation. PARP-1 also plays important roles in other disease processes, like central nervous system dysfunctions (e.g. Stroke, Parkinson), carcinogenesis and autoimmune diseases such as type I diabetes. Tregs is important for in autoimmune and inflammatory diseases.PARP-1 can affect regulatory T cell differentiation. Tregs regulate immune responses by cell-cell contact via CTLA-4.  

I agree with the comments above. In the US, most of these patients are eventually referred to pulmonary hypertension centers of excellence, and following a right heart cath, their treatment revolves around the use of sildenafil, CCB, anticoagulants, prostacyclin, bosentan, etc, in various combinations, as above-aluded to. Usually, these patients are already on some type of immunosuppressive therapy, including Plaquenil, however their use is not necessarily directed at or dictated by the PH bur rather at the underlying inflammatory disease,