A method for identification of HIV gp140 binding memory B cells in human blood

doi:10.1016/j.jim.2008.11.012

Journal of Immunological Methods

Volume 343, Issue 2, 15 April 2009, Pages 65-67

https://doi.org/10.1016/j.jim.2008.11.012 Get rights and content

Antibodies to HIV are potentially important reagents for basic and clinical studies. Historically, these reagents have been produced by random cloning of heavy and light chains in phage display libraries [Burton, D.R., Barbas, C.F. III, Persson, M.A.A., Koenig, S., Chanock, R.M., and Lerner, R.A., (1991), A large array of human monoclonal antibodies to type 1 immnodeficiency virus from combinatorial libraries of asymptomatic seropositive individuals. Proc. Natl. Acad. Sci. U. S. A. 88, 10134–10137.] and electrofusion techniques [Buchacher, A., Predl, R., Tauer, C., Purtscher, M., Gruber, G., Heider, R., Steindl, F., Trkola, A., Jungbauer, A., and Katinger, H., (1992), Human monoclonal antibodies against gp41 and gp120 as potential agent for passive immunization. Vaccines 92, 191–195]. Here we describe a method to identify and potentially enrich human memory B cells from HIV infected patients that show serum titers of neutralizing antibodies. When biotinylated gp140 is used to stain peripheral blood mononuclear cells it identifies a distinct population of gp140 binding B cells by flow cytometry.

Introduction

The HIV surface protein is composed of a trimer of gp140, which is made up of two non-covalently associated polypeptides gp120 and gp41. Antibodies to either gp120 or gp41 can have neutralizing activity in vitro and in vivo. However, in order to neutralize, antibodies need to bind to their epitope in the context of the functional trimeric GP140 (Broder et al., 1994, Yang et al., 2000). In our search for a method to identify memory B cells in the blood that bind to the surface of HIV we made use of an artificially trimerized gp140 protein that has previously been shown to resemble the native envelope spike (Yang et al., 2000).

Section snippets

Participants

HIV-1 infected patient is part of the Elite Controller Study of the Partners Aids Research Center. The patient was identified as elite controller based on clinical data (Table 1), CD4+ T cell counts and plasma viral loads below 50 RNA copies/ml in the absence of retroviral therapy (Walker, 2007). The un-infected volunteer was a 31 year old male recruited at the Rockefeller University. Human samples were collected after signed informed consent in accordance with Institutional Review Board

Results

Antigen specific IgG⁺ B cells make up a small percentage of the circulating B cell pool. These cells can be identified by their expression of CD19 and IgG as well as a high affinity to their antigen and represent somatically hypermutated post germinal center IgG memory B cells (Klein et al., 1998). The presence of such cells in blood correlates in part with serum antibody titers (Crotty et al., 2003, Leyendeckers et al., 1999, Nanan et al., 2001, Amanna et al., 2007).

To determine whether we

Discussion

Despite the importance of antibody responses to HIV there are no methods to identify, study and potentially purify circulating memory B cells that express such antibodies. We find only a low frequency of gp140 binding B cells in the IgG⁺ B cell compartment in the patient. However, the frequency of such cells was similar to the frequency of vaccinia specific circulating IgG memory B cells in vaccinia vaccinated individuals (Crotty et al., 2003). Furthermore these cells were not found in a

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Research paperA method for identification of HIV gp140 binding memory B cells in human blood

Introduction

Section snippets

Participants

Results

Discussion

Vaccine

J. Immunol. Methods

Duration of humoral immunity to common viral and vaccine antigens

N. Engl. J. Med.

Proc. Natl. Acad. Sci. U. S. A.

Cutting edge: long-term B cell memory in humans after smallpox vaccination

J. Immunol.

Research paper
A method for identification of HIV gp140 binding memory B cells in human blood