The Influence of B-cell Tolerance on Humoral Immunity to HIV-1
| dc.contributor.advisor | Zhuang, Yuan | |
| dc.contributor.author | Holl, Thomas Matthew | |
| dc.date.accessioned | 2011-01-05T14:41:45Z | |
| dc.date.available | 2012-09-01T04:30:08Z | |
| dc.date.issued | 2010 | |
| dc.department | Immunology | |
| dc.description.abstract | 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. | |
| dc.identifier.uri | ||
| dc.subject | Health Sciences, Immunology | |
| dc.subject | B cell | |
| dc.subject | B-cell Development | |
| dc.subject | B-cell Tolerance | |
| dc.subject | HIV-1 | |
| dc.subject | Humoral Immunity | |
| dc.subject | MPER | |
| dc.title | The Influence of B-cell Tolerance on Humoral Immunity to HIV-1 | |
| dc.type | Dissertation | |
| duke.embargo.months | 24 |