Dengue virus-specific, HLA-B35-restricted, human CD8+ cytotoxic T lymphocyte (CTL) clones. Recognition of NS3 amino acids 500 to 508 by CTL clones of two different serotype specificities.
Dengue virus infections are a major cause of morbidity and mortality in tropical and subtropical areas of the world. We analyzed dengue virus-specific CD8+ CD4- CTL at the clonal level to further understand the role of CD8+ CTL in dengue virus infections. Dengue virus-specific CD8+ CTL clones were established from lymphocytes of a dengue 4-immune adult. Three patterns of dengue serotype specificities were identified: 1) specific for dengue 4, 2) cross-reactive for dengue 2 and dengue 4 (subcomplex-specific); and 3) cross-reactive for all four dengue virus serotypes. Three dengue 4-specific clones and one dengue 2/dengue 4 cross-reactive clone were further analyzed. All four of the clones were HLA-B35 restricted and recognized NS3. The epitopes were mapped to amino acids (aa) 483 to 618 of NS3. The epitope was then defined by using synthetic peptides. Three dengue 4-specific clones and one dengue 2/dengue 4 cross-reactive clone recognized the same peptide (TPEGIIPTL) encompassing aa 500 to 508 of dengue 4 NS3. The peptide encompassing aa 500-508 of dengue 2 NS3 was recognized by a dengue 2/dengue 4 cross-reactive clone but was not recognized by the dengue 4-specific clones. Dengue 4-specific and dengue 2/dengue 4 cross-reactive clones used different TCR. These results indicate that CD8+ CTL clones that use different TCR and demonstrate two distinct serotype specificities recognize the same 9-mer peptide in the context of HLA-B35.
Livingston
P G
PG
Department of Medicine, University of Massachusetts Medical Center, Worcester 01655.
Kurane
I
I
Dai
L C
LC
Okamoto
Y
Y
Lai
C J
CJ
Men
R
R
Karaki
S
S
Takiguchi
M
M
Ennis
F A
FA
D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D000956:Antigens, Viral / Q000276:immunology*; D002460:Cell Line; D003601:Cytotoxicity Tests, Immunologic; D003716:Dengue Virus / Q000145:classification / Q000276:immunology*; D000939:Epitopes / Q000276:immunology; D015797:HLA-B35 Antigen / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007739:L Cells; D051379:Mice; D008969:Molecular Sequence Data; D020365:RNA Helicases; D011948:Receptors, Antigen, T-Cell / Q000235:genetics; D012697:Serine Endopeptidases; D012703:Serotyping; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014162:Transfection; D017361:Viral Nonstructural Proteins / Q000276:immunology*; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D000956:Antigens, Viral; D000939:Epitopes; D015797:HLA-B35 Antigen; D020365:RNA Helicases; D011948:Receptors, Antigen, T-Cell; D012697:Serine Endopeptidases; D017361:Viral Nonstructural Proteins; C060028:NS3 protein, flavivirus; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases