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Flow cytometry based identification of simian immunodeficiency virus Env-specific B lymphocytes.

Authors :
Fofana IB
Colantonio AD
Reeves RK
Connole MA
Gillis JM
Hall LR
Sato S
Audin CR
Evans DT
Shida H
Johnson RP
Johnson WE
Source :
Journal of immunological methods [J Immunol Methods] 2011 Jul 29; Vol. 370 (1-2), pp. 75-85. Date of Electronic Publication: 2011 Jun 13.
Publication Year :
2011

Abstract

SIV infection of macaques is the most widely employed model for preclinical AIDS vaccine and pathogenesis research. In macaques, high-titer virus-specific antibodies are induced by infection, and antibody responses can drive evolution of viral escape variants. However, neutralizing antibodies (Nabs) induced in response to SIVmac239 and SIVmac251 infection or immunization are generally undetectable or of low titer, and the identification and cloning of potent Nabs from SIVmac-infected macaques remains elusive. Based on recent advances in labeling HIV-specific B lymphocytes [1-3], we have generated recombinant, secreted, soluble SIVmac envelope (Env) proteins (gp120 and gp140) for detection and quantification of SIVmac Env-specific B lymphocytes. In contrast to HIV-1, we found that soluble SIVmac239 gp140 retains the ability to form stable oligomers without the necessity for introducing additional, stabilizing modifications. Soluble oligomeric gp140 reacted with rhesus anti-SIV Env-specific monoclonal antibodies (MAbs), and was used to deplete Env-specific antibodies with SIV neutralization capability from plasma taken from a rhesus macaque immunized with live attenuated SIVmac239∆nef. Soluble gp120 and gp140 bound to SIV-specific immortalized B cells, and to SIV Env-specific B lymphocytes in peripheral blood of immunized animals. These reagents will be useful for analyzing development of Env-specific B cell responses in preclinical studies using SIV-infected or vaccinated rhesus macaques.<br /> (Copyright © 2011 Elsevier B.V. All rights reserved.)

Details

Language :
English
ISSN :
1872-7905
Volume :
370
Issue :
1-2
Database :
MEDLINE
Journal :
Journal of immunological methods
Publication Type :
Academic Journal
Accession number :
21689659
Full Text :
https://doi.org/10.1016/j.jim.2011.05.010