Computational prediction and identification of dengue virus-specific CD4(+) T-cell epitopes.
In this study, we tried to identify dengue virus-specific CD4(+) T-cell epitopes, which can induce PBMC (peripheral blood mononuclear cells) isolated from DF convalescent patients (dengue virus type 1 infection) to secrete IFN-gamma. PBMC of DF convalescent patients were stimulated in vitro with dengue virus-derived peptides, which were prepared based on the prediction of dengue virus-specific CD4(+) T-cell epitopes by using RANKpep online software. Subsequently, the frequency of IFN-gamma producing T cells and percentage of IFN-gamma(+) CD4(+) T cells were measured by using ELISPOT assay and ICS assay (intracellular cytokine straining), respectively. The positive response of PBMC by ELISPOT showed that the numbers of SFC (spots forming cells) ranged from 50 to 310 SFC/1x10(6) PBMC. The positive response of PBMC by ICS assay showed that the percentage of IFN-gamma(+) CD4(+) T cells ranged from 0.03 to 0.27%. As a result, C(45-57) (KLVMAFIAFLRFL), E(396-408) (SSIGKMFEATARG), NS3(23-35) (YRILQRGLLGRSQ), and NS3(141-155) (NREGKIVGLYGNGVV) were identified as dengue virus-specific CD4(+) T-cell epitopes.
Department of Microbiology, Zhongshan Medical College, Sun Yat-Sen University, Guangzhou 510080, People's Republic of China.
Jin-Sheng
JS
Wen
Li-Fang
LF
Jiang
Jun-Mei
JM
Zhou
Hui-Jun
HJ
Yan
Dan-Yun
DY
Fang
D000293:Adolescent; D000328:Adult; D000595:Amino Acid Sequence; D000956:Antigens, Viral / Q000276:immunology / Q000302:isolation & purification*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D019295:Computational Biology; D003715:Dengue / Q000276:immunology / Q000821:virology; D003716:Dengue Virus / Q000145:classification / Q000276:immunology* / Q000302:isolation & purification; D018604:Epitope Mapping; D018984:Epitopes, T-Lymphocyte / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008213:Lymphocyte Activation; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology* / Q000302:isolation & purification; D012984:Software; D013045:Species Specificity
D000956:Antigens, Viral; D018984:Epitopes, T-Lymphocyte; D010455:Peptides; D007371:Interferon-gamma