Dengue virus-specific human CD4+ T-lymphocyte responses in a recipient of an experimental live-attenuated dengue virus type 1 vaccine: bulk culture proliferation, clonal analysis, and precursor frequency determination.
We analyzed the CD4+ T-lymphocyte responses to dengue, West Nile, and yellow fever viruses 4 months after immunization of a volunteer with an experimental live-attenuated dengue virus type 1 vaccine (DEN-1 45AZ5). We examined bulk culture proliferation to noninfectious antigens, determined the precursor frequency of specific CD4+ T cells by limiting dilution, and established and analyzed CD4+ T-cell clones. Bulk culture proliferation was predominantly dengue virus type 1 specific with a lesser degree of cross-reactive responses to other dengue virus serotypes, West Nile virus, and yellow fever virus. Precursor frequency determination by limiting dilution in the presence of noninfectious dengue virus antigens revealed a frequency of antigen-reactive cells of 1 in 1,686 peripheral blood mononuclear cells (PBMC) for dengue virus type 1, 1 in 9,870 PBMC for dengue virus type 3, 1 in 14,053 PBMC for dengue virus type 2, and 1 in 17,690 PBMC for dengue virus type 4. Seventeen CD4+ T-cell clones were then established by using infectious dengue virus type 1 as antigen. Two patterns of dengue virus specificity were found in these clones. Thirteen clones were dengue virus type 1 specific, and four clones recognized both dengue virus types 1 and 3. Analysis of human leukocyte antigen (HLA) restriction revealed that five clones are HLA-DRw52 restricted, one clone is HLA-DP3 restricted, and one clone is HLA-DP4 restricted. These results indicate that in this individual, the CD4+ T-lymphocyte responses to immunization with live-attenuated dengue virus type 1 vaccine are predominantly serotype specific and suggest that a multivalent vaccine may be necessary to elicit strong serotype-cross-reactive CD4+ T-lymphocyte responses in such individuals.
Department of Medicine, University of Massachusetts Medical Center, Worcester 01655.
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Green
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Kurane
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Edelman
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Tacket
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Eckels
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Vaughn
C H
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Hoke
F A
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Ennis
D000328:Adult; D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000032:analysis*; D000956:Antigens, Viral / Q000276:immunology; D017252:CD3 Complex / Q000032:analysis; D015704:CD4 Antigens / Q000032:analysis*; D002478:Cells, Cultured; D002999:Clone Cells; D003602:Cytotoxicity, Immunologic; D003716:Dengue Virus / Q000145:classification / Q000276:immunology*; D006681:HLA-D Antigens / Q000276:immunology; D006801:Humans; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000382:microbiology*; D008297:Male; D012703:Serotyping; D016176:T-Lymphocyte Subsets / Q000276:immunology*; D014613:Vaccines, Attenuated / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology*
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000956:Antigens, Viral; D017252:CD3 Complex; D015704:CD4 Antigens; D006681:HLA-D Antigens; D014613:Vaccines, Attenuated; D014765:Viral Vaccines