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Detection and Enrichment of Rare Antigen-specific B Cells for Analysis of Phenotype and Function
Published on: February 16, 2017
Evaluation of antigen-specific responses using in vitro enriched T cells
N Jones1, D Agrawal, M Elrefaei
1California Department of Health Services, 850 Marina Bay Parkway, VRDL, Richmond, CA 94804, USA.
Journal of Immunological Methods
|March 1, 2003
Summary
Bispecific monoclonal antibodies (BSMAB) effectively expand and enrich specific T cell populations for immunological studies. This method maintains specificity and offers an alternative for scarce cell sources in cellular immunology assays.
Area of Science:
- Immunology
- Cell Biology
- Virology
Background:
- Antigen-specific lymphocytes are crucial for viral infection immunity.
- Peripheral blood mononuclear cells (PBMC) are standard for effector cell studies.
- Limited cell numbers can hinder immunological research.
Purpose of the Study:
- To describe the use of bispecific monoclonal antibodies (BSMAB) for T cell expansion and enrichment.
- To evaluate the specificity and sensitivity of BSMAB-enriched T cells in cellular immunology assays.
- To assess the T cell repertoire and memory phenotype of BSMAB-expanded cells.
Main Methods:
- Utilized anti-CD3:CD8 and anti-CD3:CD4 BSMAB to enrich CD8+ and CD4+ T cells.
- Quantified T cell populations using flow cytometry (>90% purity achieved).
- Assessed HIV- and CMV-specific responses and Vbeta repertoire of enriched cells.
Main Results:
- BSMAB successfully expanded and enriched CD4+ and CD8+ T cell populations within 14 days.
- Enriched T cells showed comparable or superior sensitivity and specificity in cellular immunology assays (CMI) versus PBMC.
- Antigen-specific responses were primarily from the CD45RO+ memory effector subset.
Conclusions:
- BSMAB enables significant in vitro expansion of T cells without compromising specificity.
- This technique provides a valuable alternative for immunological assays when initial cell sources are limited.
- BSMAB-enriched T cells offer a viable and functional option for studying T cell subset function.
