Progress towards an HIV-1 vaccine has been stymied by the inability to induce a protective humoral response. Well-characterized neutralization epitopes are either poorly immunogenic (e.g., b12, 2F5, 2G12) or effectively masked on the majority of primary isolates (e.g., antibodies to V3, CD4-bd and CD4-i epitopes). New evidence suggests that even highly masked primary isolates possess sensitive neutralization targets that are recognized by autologous patient sera and occasionally by heterologous sera. This suggests that mapping the epitopes involved may identify novel targets that are capable of inducing broadly neutralizing activities. We have identified several patient sera that possess broadly neutralizing activities for primary isolates, and we have developed methods for localizing the target epitopes. We now propose to identify epitopes that mediate neutralization of both autologous and heterologous clade B and clade C Envs.
The Specific Aims are: 1- To screen a large number of patient sera obtained from both North-American and African cohorts for cross-neutralizing activities, and select samples with broad neutralizing properties for detailed characterization. 2- To map target epitopes initially by examining the neutralization sensitivities of chimeras formed between neutralization-sensitive and -resistant Envs, in which key domains were exchanged using available or engineered restriction sites. Finer mapping of the epitopes involved will be performed by exchanging smaller regions and by mutagenesis of key residues. In parallel, sensitive epitopes will be mapped immunochemically using gp120 and gp41 antigens and fusion proteins expressing various domains of sensitive Envs as blocking and immunoadsorption reagents in neutralization assays. 3- To isolate monoclonal antibodies (mAbs) with broad neutralizing activities from EBV-transformed B cell cultures obtained from these patients, and from mice immunized with Env proteins or fusion proteins expressing potential neutralization targets. Screening will be performed both with binding and direct neutralization assays. The resulting mAbs will be used to further define the structure and distribution of the epitopes involved. 4- Finally, to immunize rabbits with fusion proteins expressing novel targets. These studies will evaluate the immunogenicity of the expressed sequences and to test the neutralizing properties of antibodies induced against these immunogens. It is anticipated that these studies will define novel targets that are exposed on typical neutralization-resistant primary isolates and responsible for broad neutralization, and could lead to new vaccine approaches based on immunogens expressing combinations of such epitopes.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01AI078410-05
Application #
8260290
Study Section
Special Emphasis Panel (ZAI1-KS-I (J3))
Program Officer
Li, Yen
Project Start
2008-05-01
Project End
2013-06-30
Budget Start
2012-05-01
Budget End
2013-06-30
Support Year
5
Fiscal Year
2012
Total Cost
$483,963
Indirect Cost
$144,775
Name
University of Medicine & Dentistry of NJ
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
623946217
City
Newark
State
NJ
Country
United States
Zip Code
07107
Salomon, Aidy; Krachmarov, Chavdar; Lai, Zhong et al. (2014) Specific sequences commonly found in the V3 domain of HIV-1 subtype C isolates affect the overall conformation of native Env and induce a neutralization-resistant phenotype independent of V1/V2 masking. Virology 448:363-74
Shmelkov, Evgeny; Krachmarov, Chavdar; Grigoryan, Arsen V et al. (2014) Computational prediction of neutralization epitopes targeted by human anti-V3 HIV monoclonal antibodies. PLoS One 9:e89987
Krachmarov, Chavdar; Lai, Zhong; Honnen, William J et al. (2011) Characterization of structural features and diversity of variable-region determinants of related quaternary epitopes recognized by human and rhesus macaque monoclonal antibodies possessing unusually potent neutralizing activities. J Virol 85:10730-40
Moore, Penny L; Gray, Elin S; Sheward, Daniel et al. (2011) Potent and broad neutralization of HIV-1 subtype C by plasma antibodies targeting a quaternary epitope including residues in the V2 loop. J Virol 85:3128-41
Lynch, Rebecca M; Rong, Rong; Li, Bing et al. (2010) Subtype-specific conservation of isoleucine 309 in the envelope V3 domain is linked to immune evasion in subtype C HIV-1 infection. Virology 404:59-70
Granados-Gonzalez, Viviana; Piedrahita, Leidy Diana; Martinez-Gutierrez, Marlen et al. (2009) Neutralizing inter-clade cross-reactivity of HIV-1 V1/V2-specific secretory immunoglobulin A in Colombian and French cohorts. AIDS 23:2219-22
Rong, Rong; Li, Bing; Lynch, Rebecca M et al. (2009) Escape from autologous neutralizing antibodies in acute/early subtype C HIV-1 infection requires multiple pathways. PLoS Pathog 5:e1000594