TY - JOUR
T1 - Evaluation of antigen-specific responses using in vitro enriched T cells
AU - Jones, N.
AU - Agrawal, D.
AU - Elrefaei, M.
AU - Hanson, A.
AU - Novitsky, V.
AU - Wong, J. T.
AU - Cao, Huyen
PY - 2003/3/1
Y1 - 2003/3/1
N2 - 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 CD45RAhi and CD45ROhi cell populations were expanded with the BSMAB, selective subset depletion demonstrated that the antigen-specific T cell responses were restricted to the initial CD45ROhi 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.
AB - 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 CD45RAhi and CD45ROhi cell populations were expanded with the BSMAB, selective subset depletion demonstrated that the antigen-specific T cell responses were restricted to the initial CD45ROhi 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.
KW - Bispecific monoclonal antibody
KW - CMI
KW - T cell
UR - http://www.scopus.com/inward/record.url?scp=0037374385&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0037374385&partnerID=8YFLogxK
U2 - 10.1016/S0022-1759(02)00510-0
DO - 10.1016/S0022-1759(02)00510-0
M3 - Article
C2 - 12609540
AN - SCOPUS:0037374385
SN - 0022-1759
VL - 274
SP - 139
EP - 147
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
IS - 1-2
ER -