Evaluation of antigen-specific responses using in vitro enriched T cells

N. Jones, D. Agrawal, M. Elrefaei, A. Hanson, V. Novitsky, J. T. Wong, Huyen Cao

Research output: Contribution to journalArticlepeer-review

5 Scopus citations


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.

Original languageEnglish (US)
Pages (from-to)139-147
Number of pages9
JournalJournal of Immunological Methods
Issue number1-2
StatePublished - Mar 1 2003


  • Bispecific monoclonal antibody
  • CMI
  • T cell

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology


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