GSBS Dissertations and Theses

Approval Date

10-12-2016

Document Type

Doctoral Dissertation

Academic Program

Immunology and Microbiology

Department

Medicine

First Thesis Advisor

Shan Lu, M.D., Ph.D.

Keywords

HIV-1, HIV Infections, env Gene Products, Human Immunodeficiency Virus, Monoclonal Antibodies, HIV Envelope Protein gp120, Vaccines, Vaccination, AIDS Vaccines

Subjects

Dissertations, UMMS; HIV-1; HIV Infections; env Gene Products, Human Immunodeficiency Virus; Antibodies, Monoclonal; HIV Envelope Protein gp120; Vaccines; Vaccination; AIDS Vaccines

Abstract

Human immunodeficiency type 1 (HIV-1) is able to elicit broadly potent neutralizing antibodies in a very small subset of individuals only after several years’ infection and as a result, vaccines that elicit these types of antibodies have been difficult to design. The RV144 trial showed that a moderate protection is possible, which may correlate with antibody dependent cellular cytotoxicity (ADCC) activity. Previous studies in the Lu lab demonstrated that in an HIV-1 vaccine phase I trial, DP6-001, a polyvalent Env DNA prime-protein boost formulation, could elicit potent and broadly reactive, gp120-specific antibodies with positive neutralization activities along with multiple Fc mediated effector functions. I developed a protocol for the production and analysis of HIV-1 Env-specific human monoclonal antibodies (mAbs) isolated from these DP6-001 vaccinees. By utilizing a labeled gp120 bait to isolate Env specific B cells, paired heavy and light chain immunoglobulin (Ig) genes were cloned and allowed for the production of monoclonal antibodies with specificity for gp120. By using this protocol, 13 isolated mAbs from four DP6-001 vaccinees showed broad binding activities to gp120 proteins of diverse subtypes, both autologous and heterologous to vaccine immunogens, with mostly conformational epitopes and a few V3 and C5 specific mAbs. Equally cross-reactive Fc-mediated functional activities, including ADCC and antibody dependent cellular phagocytosis (ADCP), were present with both immune sera and isolated mAbs, confirming the induction of non-neutralizing functional antibodies by the DNA prime- protein boost vaccination. Elicitation of broadly reactive mAbs by vaccination in healthy human volunteers confirms the value of the polyvalent formulation in this HIV-1 vaccine design.

DOI

10.13028/M2W307

Rights and Permissions

Copyright is held by the author, with all rights reserved.

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