Use of retroviral vectors for the analysis of SIV/HIV-specific CD8 T cell responses

Donatella R M Negri, Viviana Buffa, Pasqualina Leone, Roberta Bona, Martina Borghi, Francesca Carlini, Fabiana De Angelis, Stefano Indraccolo, Barbara Ensoli, Andrea Cara

Research output: Contribution to journalArticlepeer-review

Abstract

CD8+ T cell responses and particularly cytotoxic T lymphocyte (CTL) activity are critical factors in controlling SHIV, SIV or HIV replication during natural infection and represent key parameters which need to be monitored during vaccine development. In order to improve the methodology for measuring CD8+ T cell responses, retroviral vectors expressing the full-length SIV-Gag or HIV-Env proteins were constructed and used to transduce B lymphoblastoid cell lines (BLCL) from cynomolgus monkeys infected with SHIV89.6P. Continuous expression of Gag and Env proteins was detected in stably transduced BLCL, which induced Gag- or Env-specific T cell responses, as measured by both IFNγ-ELISPOT and chromium release assays, upon in vitro stimulation of PBMC from the SHIV89.6P-infected monkeys. Moreover, induction of Gag-specific CTL using BLCL transduced with retroviral vector expressing the SIV-Gag protein was more efficient and specific compared to that obtained using BLCL infected with a recombinant vaccinia virus (rVV) encoding for the same antigen. Assays on purified CD4+ and CD8+ T cells indicated that both populations specifically produced IFNγ, but only the CD8+ T cells mediated Gag- and Env-specific cytotoxicity, indicating preferential expansion of these effector cells. Thus, this method represents an alternative tool for the analysis of CTL responses during vaccination protocols in those animal models where little information is available on MHC class I alleles or CTL epitopes.

Original languageEnglish
Pages (from-to)153-163
Number of pages11
JournalJournal of Immunological Methods
Volume291
Issue number1-2
DOIs
Publication statusPublished - Aug 2004

Keywords

  • AIDS
  • CTL
  • Retroviral vector
  • SHIV/SIV
  • Vaccine

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

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