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Journal of Virology, January 2008, p. 428-434, Vol. 82, No. 1
0022-538X/08/$08.00+0     doi:10.1128/JVI.00638-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Quantitation of HLA Proteins Incorporated by Human Immunodeficiency Virus Type 1 and Assessment of Neutralizing Activity of Anti-HLA Antibodies{triangledown}

Samir K. Lakhashe,1 Madhuri R. Thakar,1 K. E. Bharucha,2 and Ramesh S. Paranjape1*

National AIDS Research Institute,1 B. J. Medical College, Pune, India2

Received 26 March 2007/ Accepted 8 October 2007

Human anti-human leukocyte antigen (HLA) antibodies were assessed for neutralizing activity against human immunodeficiency virus type 1 (HIV-1) carrying HLA alleles with matching specificity. Multiparous women carrying anti-HLA antibodies were identified. Plasma samples from those women were confirmed as having antibodies that specifically bound to HLA proteins expressed on the peripheral blood mononuclear cells (PBMCs) of their husbands. A primary HIV-1 isolate was cultured in the husband's PBMCs so that the virus carried matching HLA alleles. To determine the HIV-1-neutralizing activity of anti-HLA antibodies, the infectivity of the virus for GHOST cells (which express green fluorescent protein after HIV infection) was investigated in the presence of a plasma sample positive for the respective anti-HLA antibody. A neutralization assay was also performed using purified immunoglobulin G (IgG) from two plasma samples, and two plasma samples were investigated in the presence of complement. The prerequisite for anti-HLA antibody-mediated neutralization is incorporation of HLA proteins by HIV-1. Therefore, the extent of incorporation of HLA proteins by the primary HIV-1 isolate was estimated. The ratios of HLA class I protein to HIV-1 capsid (p24) protein cultured in the PBMCs of two healthy individuals were 0.017 and 0.054. These ratios suggested that the HIV-1 strain used in the assay incorporated more HLA proteins than gp160 trimers. Anti-HLA antibody-positive plasma was found to contain antibodies that specifically reacted to HIV-1 carrying cognate HLA alleles. However, incubation of HIV-1 with anti-HLA antibody- positive plasma or purified IgG did not show a reduction in viral infectivity. HIV-1-neutralizing activity was also not detected in the presence of complement. This study shows that HIV-1 primary isolates cultured in PBMCs contain significant amounts of HLA proteins. However, the binding of antibodies to those HLA proteins does not mediate a reduction in viral infectivity.

* Corresponding author. Mailing address: National AIDS Research Institute, G-73, MIDC, Bhosari, Pune, India 411 026. Phone: 91 20 27121210. Fax: 91 20 27121071. E-mail: rparanjape{at}nariindia.org

{triangledown} Published ahead of print on 17 October 2007.

Journal of Virology, January 2008, p. 428-434, Vol. 82, No. 1
0022-538X/08/$08.00+0     doi:10.1128/JVI.00638-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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