title: Evaluation of antigen-specific responses using in vitro enriched T cells.
abstract: Antigen-specific lymphocytes are important in the immune response to viral infection. Peripheral blood mononuclear cells (PBMC) are traditionally used as a source of effector cells in most immunological studies. We described here the use of the bispecific monoclonal antibodies (BSMAB) anti CD3:CD8 (CD3,8) and anti CD3:CD4 (CD3,4B) to expand and selectively enrich CD4+ and CD8+ T cells populations, respectively. The expanded cells demonstrated >90% CD3+CD4+ or CD3+CD8+ by 14 days. We measured HIV- and CMV-specific responses of these subset-enriched T cell and found that sensitivity and specificity is similar or higher when compared to PBMC in various cellular immunology assays (CMI). Vbeta analysis of BSMAB-enriched cells demonstrated comparable repertoire to the parent PBMC. Although both CD45RA(hi) and CD45RO(hi) cell populations were expanded with the BSMAB, selective subset depletion demonstrated that the antigen-specific T cell responses were restricted to the initial CD45RO(hi) memory effector subgroup. In conclusion, BSMAB in vitro enrichment of T cells allows significant expansion of the cell population without loss of specificity. This technique of cell expansion permits studies of T cell subset function in situations where the initial cell source is scarce, and presents an alternative for viable and functional T cells in immunological assays.
authors.affiliation: California Department of Health Services, 850 Marina Bay Parkway, VRDL, Richmond, CA 94804, USA.
authors.forename: N
authors.initials: N
authors.lastname: Jones
authors.forename: D
authors.initials: D
authors.lastname: Agrawal
authors.forename: M
authors.initials: M
authors.lastname: Elrefaei
authors.forename: A
authors.initials: A
authors.lastname: Hanson
authors.forename: V
authors.initials: V
authors.lastname: Novitsky
authors.forename: J T
authors.initials: JT
authors.lastname: Wong
authors.forename: Huyen
authors.initials: H
authors.lastname: Cao
mesh_terms: D018033:Antibodies, Bispecific / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000956:Antigens, Viral / Q000276:immunology*; D017252:CD3 Complex / Q000276:immunology; D015704:CD4 Antigens / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D016827:CD8 Antigens / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002460:Cell Line; D003587:Cytomegalovirus / Q000276:immunology; D006678:HIV / Q000276:immunology; D015658:HIV Infections / Q000276:immunology; D006801:Humans; D016130:Immunophenotyping; D007700:Kinetics; D008213:Lymphocyte Activation; D016176:T-Lymphocyte Subsets / Q000145:classification / Q000276:immunology*
chemical_list: D018033:Antibodies, Bispecific; D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D017252:CD3 Complex; D015704:CD4 Antigens; D016827:CD8 Antigens