Biochem Paper - Christina Lavelle Generation and...

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Christina Lavelle Generation and Characterization of Neutralizing Human Monoclonal Antibodies against Human Immunodeficiency Virus Type 1 Tat Antigen The structural and regulatory proteins that are essential for the pathogenesis of AIDS are encoded for by the Human immunodeficiency virus type 1 (HIV-1). Essential for the transcription of HIV-1 and viral replication, Tat is a small nuclear protein made of approximately 86- to 101- amino acids. It is released in vitro by infected cells and immediately inhibits many immunosuppressive functions, such as production of interleukin-12 by human peripheral blood mononuclear cells (PBMCs). The purpose of the experiment described in this paper was to determine if immunoglobulin M antibodies might provide an initial defense against the effects of the Tat antigen after HIV infection by inhibiting Tat-induced transactivation and viral replication. The study took place using PBMCs purified from blood samples from two healthy HIV-negative volunteers (called J and G) and one HIV-1-seropositive patient (called B) who were all immunized with Tat toxoid. A DNA library of each sample was made and clones were extracted to be tested. All three samples showed a high antibody count to Tat and an inhibition to Tat-mediated transactivation, however, only samples from J and B were able to produce antibodies reactive in enzyme-linked immunosorbent assay (ELISA) with recombinant Tat (rTat). These were called IgG J3B2 and IgG B1E3.
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Biochem Paper - Christina Lavelle Generation and...

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