Multiple sclerosis (MS) is the most common demyelinating disease of humans. Although its cause is unknown, clues to the nature of disease may be found in the increased IgG and oligoclonal bands (OGBs) present in the CSF and brain of MS patients, an phenomenon almost exclusively seen with infectious and inflammatory diseases of the CNS. Although the specificity of OGBs in MS is unknown, in infectious diseases of the CNS, OGBs are specific for the agent that causes disease, thus providing a rationale for our hypothesis that OGBs in MS brain and CSF are directed against the antigen that causes disease. In the past funding period, analysis of the lgG heavy (VH) and light (VL) chain variable region sequences expressed in acute MS brains and CSFs revealed a restricted IgG repertoire indicative of an antigen-driven B cell response. To produce recombinant antibodies from prominent MS Ig sequences that accurately duplicate the B cell heavy- and light- antibody chain pairings found in vivo, we developed an RT-PCR protocol to amplify the expressed V-region sequences of single B cells and plasma cells isolated from MS CSF by fluorescence-activated cell sorting. The paired heavy and light chains from CSF clonal B cell populations will now be used to produce a panel of recombinant mAbs whose specificity will be determined. Once specificity is established, cDNAs whose protein products react with MS-specific Abs will be cloned and characterized. Also, to better understand the dynamics of the B cell response in MS and to further characterize the nature of the humoral response in MS, changes in the IgG repertoire of MS CSF with disease progression will be analyzed. We are well-prepared to conduct these studies because we have: (a) a well-characterized pool of CSF from MS patients and patients with other CNS inflammatory disease; (b) a demonstrated ability to amplify disease-relevant IgG sequences from human CSF and brain, and to use these sequences to generate recombinant IgG; and (c) the expertise in molecular biology and immunology to identify the antigenic target of the recombinant Abs. Identification of an MS-specific antigen(s) will have wide application, not only for early definitive diagnosis, but also for developing strategies to modulate and possibly prevent disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Program Projects (P01)
Project #
5P01NS032623-20
Application #
7561080
Study Section
National Institute of Neurological Disorders and Stroke Initial Review Group (NSD)
Project Start
Project End
Budget Start
2008-02-01
Budget End
2009-01-31
Support Year
20
Fiscal Year
2008
Total Cost
$615,757
Indirect Cost
Name
University of Colorado Denver
Department
Type
DUNS #
041096314
City
Aurora
State
CO
Country
United States
Zip Code
80045
Cohrs, Randall J; Lee, Katherine S; Beach, Addilynn et al. (2017) Targeted Genome Sequencing Reveals Varicella-Zoster Virus Open Reading Frame 12 Deletion. J Virol 91:
Ouwendijk, Werner J D; van Veen, Suzanne; Mahalingam, Ravi et al. (2017) Simian varicella virus inhibits the interferon gamma signalling pathway. J Gen Virol :
Ouwendijk, Werner J D; Getu, Sarah; Mahalingam, Ravi et al. (2016) Characterization of the immune response in ganglia after primary simian varicella virus infection. J Neurovirol 22:376-88
Birlea, Marius; Owens, Gregory P; Eshleman, Emily M et al. (2013) Human anti-varicella-zoster virus (VZV) recombinant monoclonal antibody produced after Zostavax immunization recognizes the gH/gL complex and neutralizes VZV infection. J Virol 87:415-21
Mueller, Niklaus H; Bos, Nathan L; Seitz, Scott et al. (2012) Recombinant monoclonal antibody recognizes a unique epitope on varicella-zoster virus immediate-early 63 protein. J Virol 86:6345-9
Nagel, Maria A; Bert, Robert J; Gilden, Don (2012) Raeder syndrome produced by extension of chronic inflammation to the internal carotid artery. Neurology 79:1296-7
Wolf, James; Nagel, Maria A; Mahalingam, Ravi et al. (2012) Chronic active varicella zoster virus infection. Neurology 79:828-9
Brennan, Kathryn M; Galban-Horcajo, Francesc; Rinaldi, Simon et al. (2011) Lipid arrays identify myelin-derived lipids and lipid complexes as prominent targets for oligoclonal band antibodies in multiple sclerosis. J Neuroimmunol 238:87-95
Azarkh, Yevgeniy; Gilden, Don; Cohrs, Randall J (2010) Molecular characterization of varicella zoster virus in latently infected human ganglia: physical state and abundance of VZV DNA, Quantitation of viral transcripts and detection of VZV-specific proteins. Curr Top Microbiol Immunol 342:229-41
Sargsyan, S A; Shearer, A J; Ritchie, A M et al. (2010) Absence of Epstein-Barr virus in the brain and CSF of patients with multiple sclerosis. Neurology 74:1127-35

Showing the most recent 10 out of 146 publications