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dc.contributor.advisor Zhuang, Yuan en_US
dc.contributor.author Holl, Thomas Matthew en_US
dc.date.accessioned 2011-01-05T14:41:45Z
dc.date.available 2012-09-01T04:30:08Z
dc.date.issued 2010 en_US
dc.identifier.uri http://hdl.handle.net/10161/3053
dc.description Dissertation en_US
dc.description.abstract <p>Several HIV-1 neutralizing antibodies (e.g. 2F5, 4E10) have been shown to react with self-antigens, suggesting that effective humoral responses to HIV-1 may be constrained by the tolerization of HIV-reactive B cells that also recognize self-antigens. I have tracked the development of 2F5-like HIV-1 gp41 membrane proximal external region (MPER)-reactive B cells throughout ontogeny using B-cell tetramer reagents. In BL/6 mice, MPER-binding populations are lost during normal B-cell development and immunization with HIV-1 MPER antigen does not elicit robust humoral responses. I have identified Kynureninase as a self-antigen that is recognized by 2F5 antibody and, therefore, is a molecule that could mediate the developmental loss of B cells reactive to an epitope shared by HIV gp41 and Kynureninase. To recover these MPER-reactive cells, I describe and characterize a stromal-cell independent culture system that efficiently supports pro-B cell to IgM+ B-cell development with near normal levels of IgH and Igkappa diversity. B-cell development in vitro closely follows the patterns of development in vivo with culture derived (CD) B cells demonstrating characteristic patterns of surface antigen expression and gene activation. Immature and transitional B-cell compartments are reduced, due to the induction of tolerance, in the bone marrow of 3H9 IgH knockin mice ; however, cultures of 3H9 IgH knockin pro-B cells yields high frequencies of "forbidden", autoreactive IgM+ B cells. Furthermore, serum IgG autoantibody exceeded that present in autoimmune, C4-/- animals following the reconstitution of RAG-1-/- mice with IgM+ CD cells derived from BL/6 mice. I show that HIV-1 MPER-reactive B cells are recovered from both BL/6 and 2F5 IgH knockin bone marrow using this in vitro culture system. RAG-1-/- mice reconstituted with these culture-derived B and T cells generate strong germinal center and antibody responses to HIV-1 MPER antigens. These data demonstrate that the humoral immune response to this HIV-1 gp41 MPER antigen can be restored in mice when the constraints of B-cell tolerance have been relaxed.</p> en_US
dc.subject Health Sciences, Immunology en_US
dc.subject B cell en_US
dc.subject B-cell Development en_US
dc.subject B-cell Tolerance en_US
dc.subject HIV-1 en_US
dc.subject Humoral Immunity en_US
dc.subject MPER en_US
dc.title The Influence of B-cell Tolerance on Humoral Immunity to HIV-1 en_US
dc.type Dissertation en_US
dc.department Immunology en_US
duke.embargo.months 24 en_US

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