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

J Immunol Methods. 2009 Apr 15;343(2):65-7. doi: 10.1016/j.jim.2008.11.012. Epub 2008 Dec 25.

Abstract

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 immunodeficiency 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.

MeSH terms

  • Adult
  • B-Lymphocytes / immunology*
  • B-Lymphocytes / virology
  • Biotinylation
  • Flow Cytometry
  • HIV Antibodies / blood*
  • HIV Antibodies / immunology
  • HIV Infections / blood
  • HIV Infections / immunology
  • HIV-1 / immunology*
  • Humans
  • Immunologic Memory*
  • Male
  • Neutralization Tests
  • env Gene Products, Human Immunodeficiency Virus / metabolism*

Substances

  • HIV Antibodies
  • env Gene Products, Human Immunodeficiency Virus
  • gp140 envelope protein, Human immunodeficiency virus 1