This revised application is in response to the Program Announcement PA-09-119, Phased Innovation Award Program for AIDS Vaccine Research (R21/R33). This application proposes to produce neutralizing human monoclonal antibody (mAb) from human volunteers who received a polyvalent DNA prime-protein boost HIV vaccine in a Phase I clinical trial and their sera have demonstrated positive neutralizing activities against primary HIV-1 from clades A to E. If successful, this will be the first time that neutralizing human mAbs can be produced from HIV vaccinees. Detailed analyses of these mAbs will provide valuable information on their specificity, strength, and breadth of neutralizing activities. Similar to several available mAbs generated from HIV-1 infected patients, successful production of human mAb from HIV-1 vaccinees will provide useful information for the further optimization of Env-based HIV-1 vaccines.
Specific Aims for R21 phase (Years 1~2):
Specific Aim 1 : To produce human hybridoma using PBMCs isolated from human vaccinees and to screen and identify Env-specific monoclonal antibodies including those with neutralizing activities.
Specific Aim 2 : To generate mAbs from single Env-specific B cells using a high throughput in vitro Ig expression strategy Specific Aims for R33 phase (Years 3-5):
Specific Aim 3 To examine the strength and breadth of neutralizing mAbs and identify other protective functions of non-neutralizing mAbs produced from our proposed studies.
Specific Aim 4 : To analyze the epitope specificity of neutralizing mAbs produced in this proposal.
Specific Aim 5 : To expand the production of Env-specific mAbs from additional vaccinees including PBMCs collected at different time points in the immunization schedule.
We propose to use existing blood cells collected from human volunteers who received a novel HIV vaccine in an already completed clinical trial to generate a unique type of cells that can continuously produce highly specific antibodies - research tools that can guide the design of next generation of HIV vaccines.
|Costa, Matthew R; Pollara, Justin; Edwards, Regina Whitney et al. (2016) Fc Receptor-Mediated Activities of Env-Specific Human Monoclonal Antibodies Generated from Volunteers Receiving the DNA Prime-Protein Boost HIV Vaccine DP6-001. J Virol 90:10362-10378|
|Zhang, Lu; Wang, Wei; Wang, Shixia (2015) Effect of vaccine administration modality on immunogenicity and efficacy. Expert Rev Vaccines 14:1509-23|
|Hollister, Kristin; Chen, Yuxin; Wang, Shixia et al. (2014) The role of follicular helper T cells and the germinal center in HIV-1 gp120 DNA prime and gp120 protein boost vaccination. Hum Vaccin Immunother 10:1985-92|
|Chen, Yuxin; Vaine, Michael; Wallace, Aaron et al. (2013) A novel rabbit monoclonal antibody platform to dissect the diverse repertoire of antibody epitopes for HIV-1 Env immunogen design. J Virol 87:10232-43|
|Wang, Shixia; Kishko, Michael; Wan, Shengqin et al. (2012) Pilot study on the immunogenicity of paired Env immunogens from mother-to-child transmitted HIV-1 isolates. Hum Vaccin Immunother 8:1638-47|
|Vaine, Michael; Wang, Shixia; Liu, Qin et al. (2010) Profiles of human serum antibody responses elicited by three leading HIV vaccines focusing on the induction of Env-specific antibodies. PLoS One 5:e13916|
|Lu, Shan; Grimes Serrano, Jill M; Wang, Shixia (2010) Polyvalent AIDS vaccines. Curr HIV Res 8:622-9|