by Cort Johnson | Mar 21, 2017
A Different Look at the Immune System
The ultimate goal of this work is the development of a non-subjective clinical tool for diagnosing patients with ME. Lombardi et. al.
Last month Health Rising looked at a potential ion channel biomarker coming out of the NCNED in Australia. This month brings yet another potential biomarker hailing from the Nevada Center for Biomedical Research (formerly WPI) at the University of Reno. Time will tell if either turns out. In the meantime it’s good to see ME/CFS research centers using innovative techniques to look at this disease in entirely new ways.
Immunosignatures use antibody activity to get an idea of what the immune system is reacting to. The idea is that B-cells are involved in some way in most immune responses. Finding which peptides or antigens the antibodies in a person’s blood bind to could tell us what the immune system is reacting to. If ME/CFS is, at least in part, an immune disease, immunosignature research could tell us much. This approach has been used to provide diagnostic biomarkers for cancer, Valley fever, Alzheimer’s disease and others.
Talk about casting a large net. First antibodies are incubated with or exposed to thousands or in the case of this study over 100,000 randomly generated peptides (short-chains of amino acids). Because so many different peptides are used, the immunosignatures produced by this technique can be highly sensitive (i.e. highly accurate in identifying patients). They’re far more sensitive than say an ELISA blood test that might sum the contributions of several antibodies.
The technology used to produce immunosignatures is also very robust; a single drop of blood blotted on some paper and sent through the mail can suffice. It’s also potentially much more stable than the cytokine studies that have, at times, been so variable in ME/CFS. Because antibodies are much more stable than cytokines (they last longer in the body), they might yield better results. Read More
Physicians lack effective tools to aid in the diagnosis and treatment of those who are impacted by ME/CFS, a complex and often disabling disease. That could change now that scientists at Nevada Center for Biomedical Research (NVCBR) have taken the first step in developing a clinical assay for diagnosing ME/CFS patients through the creation of an antibody immune signature.
The work was conducted in collaboration with Drs. Stephen Johnston and Phillip Stafford, at Arizona State University’s Biodesign Institute, Innovations in Medicine, and Drs. Karen Schlauch and Richard Tillet, from the University of Nevada, Reno, as well as other researchers from around the world.
The group’s findings recently appeared in the prestigious journal Molecular Neurobiology https://link.springer.com/article/10.1007/s12035-016-0334-0
The immune system produces proteins called antibodies that bind to the surface of pathogens, such as viruses and bacteria, to neutralize the pathogens. Occasionally, this system becomes dysregulated and produces antibodies to our own tissue, resulting in autoimmunity. Utilizing a microchip comprised of thousands of small random protein sequences (referred to as random peptides), researchers at NVCBR, in collaboration with the scientists at ASU’s Biodesign Institute, screened blood sera from ME/CFS cases and healthy controls, from two geographic distinct cohorts, and identified a diagnostic pattern of antibody/peptide binding that identifies ME/CFS patients with high specificity and sensitivity.
“The purpose of screening sera with a random peptide array is to identify things that our body sees as foreign such as the molecules that make up pathogens or the proteins of our own tissue, in the context of autoimmunity,” explained Dr. Vincent Lombardi, NVCBR Research Director. “Now that we have identified a group of random peptides that differentially bind with the antibodies of ME/CFS patients, our next challenge is to figure out what antigens these random peptides represent in the real world. Much can be learned about the causes of this disease once the antigens are accurately identified.”
Dr. Lombardi originally conceived of the project with the aim of identifying potential pathogens associated with ME/CFS as well as autoantigens. In addition to identifying prospective pathogen-associated antigens and autoantigens, the data also produced an immunosignature that can accurately identify ME/CFS patients from healthy controls. Future in-depth analysis of these data will help researchers determine which antigens are most relevant to the disease process and whether or not there are existing treatments available to target the implicated biochemical pathways.
Going forward, NVCBR researchers intend to confirm the work using a larger ME/CFS cohort as well as other disease cohorts with overlapping symptomology such as multiple sclerosis. The immunosignature technology, developed by ASU, has been used successfully with other neuroimmune diseases including Alzheimer’s disease.
“The goal of our research program is to ascertain the underlying mechanisms of neuroimmune disease as well as identify disease-related biomarkers and more effective treatments for the patients,” stated Dr. Lombardi. “A robust immunosignature of ME/CFS would help achieve these goals.”
The National Institutes of Health (NIH) issued a request for Center grant proposals, with a promise to fund two to three regional ME/CFS Collaborative Research Centers as early as September 2017. The NVCBR research team and their collaborators are working hard to create a competitive proposal based on the knowledge that they have gained through their many revealing studies.
“We are excited to have the opportunity to apply for major federal research support on behalf of all of those who suffer the disabling effects of ME/CFS,” stated Annette Whittemore, President and CEO of NVCBR. “We have always hoped that the federal government would support comprehensive research centers across America. We applaud the National Institutes of Health, particularly Director Dr. Francis Collins and Program Director Dr. Vicky Whittemore, for their commitment to funding comprehensive ME/CFS research centers in addition to other promising ME/CFS research proposals.”
The plan appears to be similar to the ACE (Autism Centers of Excellence) program, which currently comprises three research centers and eight research networks around the United States. Networks such as these have been instrumental to research progress in many other diseases. They can help shorten the time it takes to biologically define a disease and encourage pharmaceutical research and development of effective treatments, something that patients with ME/CFS have been seeking for decades.
“The overarching goal of this initiative is to establish a network of Centers that will work collaboratively to define the cause(s) of and discover improved treatments for ME/CFS,” according to the funding opportunity announcement. “A more immediate goal for each Center is to rapidly advance synergistic, interdisciplinary research programs while serving as local resources and national leaders in ME/CFS research.”
The Centers will be funded by various NIH components, with funding limits for each Center set at 1.2 million dollars per year for up to 5 years. This is a major improvement over previous years in which federal funding was limited to specific research proposals. This increase in a federal commitment to ME/CFS research might also lead to pharmaceutical support of treatment trials within a few years, something that cannot happen soon enough for the millions who are disabled and without treatment.
You can read the entire funding opportunity notice at https://grants.nih.gov/grants/guide/rfa-files/RFA-NS-17-021.html
Jen Brea gives an impassioned TED talk about the realities of dealing with the disease known as ME or myalgic encephalomyelitis. Please listen as she tells about her struggles from the effects of the disease and the difficulties she encountered trying to find a doctor who could diagnose and treat her debilitating illness. Sharing this talk with your friends can help them understand the serious impact this illness has on the lives of so many and why Nevada Center for Biomedical Research needs your support to help us find answers to this complex disease.
The Skin–Brain Connection Hypothesis, Bringing Together CCL27-Mediated T-Cell Activation in the Skin and Neural Cell Damage in the Adult Brain
Authors: Nataliya L. Blatt, Timur I. Khaiboullin,Vincent C. Lombardi, Albert A. Rizvanov and Svetlana F. Khaiboullina1,
Publication: Front. Immunol., 16 January 2017
Recent discovery of an association of low serum melatonin levels with relapse in multiple sclerosis (MS) opens a new horizon in understanding the pathogenesis of this disease. Skin is the main organ for sensing seasonal changes in duration of sunlight exposure. Level of melatonin production is dependent on light exposure. The molecular mechanisms connecting peripheral (skin) sensing of the light exposure and developing brain inflammation (MS) have not been investigated. We hypothesize that there is a connection between the reaction of skin to seasonal changes in sunlight exposure and the risk of MS and that seasonal changes in light exposure cause peripheral (skin) inflammation, the production of cytokines, and the subsequent inflammation of the brain. In skin of genetically predisposed individuals, cytokines attract memory cutaneous lymphocyte-associated antigen (CLA+) T lymphocytes, which then maintain local inflammation. Once inflammation is resolved, CLA+ lymphocytes return to the circulation, some of which eventually migrate to the brain. Once in the brain these lymphocytes may initiate an inflammatory response. Our observation of increased CC chemokine ligand 27 (CCL27) in MS sera supports the involvement of skin in the pathogenesis of MS. Further, the importance of our data is that CCL27 is a chemokine released by activated keratinocytes, which is upregulated in inflamed skin. We propose that high serum levels of CCL27 in MS are the result of skin inflammation due to exposure to seasonal changes in the sunlight. Future studies will determine whether CCL27 serum level correlates with seasonal changes in sunlight exposure, MS exacerbation, and skin inflammation.
Humoral Immunity Profiling of Subjects with MyalgicEncephalomyelitis Using a Random Peptide MicroarrayDifferentiates Cases from Controls with High Specificityand Sensitivity
Authors: Sahajpreet Singh, Phillip Stafford, Karen A. Schlauch, Richard R. Tillett, Martin Gollery, Stephen Albert Johnston, Svetlana F. Khaiboullina, Kenny L. De Meirleir, Shanti Rawat, Tatjana Mijatovic, Krishnamurthy Subramanian, András Palotás, Vincent C. Lombardi
Publication: Mol Neurobiol (DOI 10.1007/s12035-016-0334-0)
Abstract Myalgic encephalomyelitis (ME) is a complex, heterogeneous illness of unknown etiology. The search for biomarkers that can delineate cases from controls is one of the most active areas of ME research; however, little progress has been made in achieving this goal. In contrast to identifying biomarkers that are directly involved in the pathological process, an immunosignature identifies antibodies raised to proteins expressed during, and potentially involved in, the pathological process. Although these proteins might be unknown, it is possible to detect antibodies that react to these proteins using random peptide arrays. In the present study, we probe a custom 125,000 random 12-mer peptide microarray with sera from 21 ME cases and 21 controls from the USA and Europe and used these data to develop a diagnostic signature. We further used these peptide sequences to potentially uncover the naturally occurring candidate antigens to which these antibodies may specifically react with in vivo. Our analysis revealed a subset of 25 peptides that distinguished cases and controls with high specificity and sensitivity. Additionally, Basic Local Alignment Search Tool (BLAST) searches suggest that these peptides primarily represent human selfantigens and endogenous retroviral sequences and, to a minor extent, viral and bacterial pathogens.