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Updated: Aug 13, 2026
Generation of Multivirus-specific T Cells to Prevent/treat Viral Infections after Allogeneic Hematopoietic Stem Cell Transplant
Published on: May 27, 2011
Dengue virus-specific, human CD4+ cytotoxic T lymphocytes generated in short-term culture
J Zivny1, I Kurane, C O Tacket
1Department of Medicine, University of Massachusetts Medical Center, Worcester.
Insights
Dengue virus-specific CD4+ CD8- cytotoxic T lymphocytes (CTL) are generated in short-term cultures. These CD4+ CTL recognize dengue 1 antigen and are restricted by HLA class II molecules, suggesting their role during dengue infection.
Area of Science:
- Immunology
- Virology
- Cellular Biology
Background:
- Previous studies reported cytotoxic activity of dengue virus-specific CD4+ CD8- T cell clones in long-term cultures.
- The presence of these cells in short-term bulk cultures remained to be determined.
Purpose of the Study:
- To investigate the generation and characteristics of dengue virus-specific CD4+ CD8- cytotoxic T lymphocytes (CTL) in short-term bulk cultures.
Main Methods:
- Peripheral blood mononuclear cells (PBMC) from a dengue-vaccinated donor were cultured with dengue 1 antigen.
- Cytotoxic activity against antigen-pulsed target cells was assessed.
- Immunophenotyping using monoclonal antibodies and HLA restriction analysis were performed.
Main Results:
- PBMC cultured with dengue 1 antigen exhibited specific cytotoxic activity against autologous cells pulsed with the same antigen.
- The cytotoxic activity was mediated by CD4+ T cells, as it was abrogated by anti-CD4 antibody treatment but not by anti-CD8.
- These CD4+ CTL were detected as early as day 5 and were restricted by HLA class II molecules (HLA-DQw1 and HLA-DRw52).
Conclusions:
- Dengue virus-specific CD4+ CD8- CTL are generated in short-term bulk cultures, similar to long-term cultures.
- These findings support the concept that CD4+ CTL play a role in vivo during dengue virus infection.
Abstract:
We previously reported cytotoxic activity of dengue virus-specific CD4+ CD8- T cell clones established in long-term in vitro culture. In the present experiments we tried to determine whether dengue virus-specific CD4+ CD8- CTL3 are present in short-term bulk cultures. Peripheral blood mononuclear cells (PBMC)3 from a donor who had been immunized with an experimental live attenuated dengue 1 vaccine 8 months earlier were used. PBMC were incubated with noninfectious dengue 1 antigen (Ag)3 for 7 days, and were examined for dengue 1-specific cytotoxic activity. PBMC cultured with dengue 1 Ag lysed autologous lymphoblastoid cell line (LCL)3 pulsed with noninfectious dengue 1 Ag, but did not lyse LCL pulsed with Ag of other dengue serotype, West Nile virus, or yellow fever virus, or control Ag. Treatment of cultured PBMC with monoclonal antibody to CD3 or CD4 and complement abrogated the cytotoxic activity but treatment with a monoclonal antibody to CD8 and complement did not. A time course study showed that dengue 1 Ag-specific CTL were first detected in 5 day cultures. Lysis of target cells by these CD4+ CTL were restricted by HLA class II, and HLA DQw1 and HLA DRw52 were determined to be the restriction molecules. These results indicate that dengue virus-specific CD4+ CD8- CTL are generated in short-term bulk cultures as well as in long-term-cultured cell lines, and support the concept that CD4+ CTL may be generated in vivo during infection.
