Selective binding of CD4 and CD8 T-cells to antigen presenting cells for enrichment of CMV and HIV specific T-lymphocytes

Giuseppina Li Pira, Federico Ivaldi, Fabrizio Manca

Research output: Contribution to journalArticle

Abstract

Adherent antigen presenting cells (APC) pulsed with protein or peptide antigens were used to capture specific CD4 or CD8 T-cells derived from established T-cell lines or from PBMC of immune subjects based on physiological interaction between TCR and MHC-peptide complex. This method could be applied independently of epitope specificity, HLA restriction alleles, activation markers and secreted cytokines, parameters required by other methods for selection of specific T cells. Non specific T-cells were removed by applying a 1. g force that did not affect binding of specific T-lymphocytes. Lymphocyte selection was specific and the average recovery was 36% for CD4 T-cells. CD8 T-cells proved trickier to purify, since solid phase APC were recognized as targets for cytotoxicity. Specificity was comparable to CD4 cells, but the average recovery for CD8 cells was 26%. No residual alloreactivity was detected in expanded T-cells. Frequency and recovery of specific T-cells were comparable to other current technologies, such as generation of T-cell lines and cytokine capture method. Since antigen and IL2 are the only reagents added to the cultures, this physiological procedure can be proposed for selection and expansion of pathogen specific T-cells not only for research purposes, but also for adoptive reconstitution of immunocompromised subjects if performed under GMP conditions.

Original languageEnglish
Pages (from-to)125-131
Number of pages7
JournalJournal of Immunological Methods
Volume376
Issue number1-2
DOIs
Publication statusPublished - Feb 28 2012

Keywords

  • APC/T-cell binding
  • Cell selection
  • Cellular therapy
  • CMV
  • Specific T-cell

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy

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