Identification of human leucocyte antigen -A* 0201-restricted hepatitis C virus-cytotoxic T lymphocyte epitopes / 中华传染病杂志

Objective To identify human leucocyte antigen (HLA)-A* 0201-restricted hepatitis C virus (HCV)-cytotoxic T lymphocyte (CTL) epitopes. Methods Based on the prediction results of RANKpep and SYFPEITHI prediction programs, six candidate CTL epitopes were selected and synthesized. The affinity of candidate CTL epitopes to HLA-A* 0201 molecules of T2 cells was explored. Subsequently, enzyme-linked immunosorbent spot (ELISPOT) assay and intracellular cytokine staining (ICS) were utilized to determine whether candidate CTL epitopes could induce the recall positive response in peripheral blood mononuclear cells (PBMC) of HLA-A* 0201 positive HCV-1b-infected patients. Results Among six candidate CTL epitopes, peptides C_181(LLSCLTTPV) and NS2_172 (VLQAGLIRV) had high affinity to HLA-A* 0201 molecules. Moreover, the affinity was proportional to the concentration of peptide. Furthermore, among ten HLA-A* 0201 positive HCV-1b-infected patients, the frequencies of C_181 and NS2_172-specific interferon (IFN)-γ-producing cells were 0-19 spots forming cells (SFC)/1 × 105 PBMC and 0-20 SFC/1 × 105 PBMC, respectively.The percentages of C_ 181 and NS2_172-specific IFN-γ+ CD8+ T lymphocytes in total CD8+ T lymphocytes were 0.006%-0.065% and 0.005%-0.080%, respectively. Conclusion Peptides C_181 (LLSCLTTPV) and NS2_172 (VLQAGLIRV) are identified as novel HLA-A* 0201-restricted HCV-CTL epitopes.

Identification of hepatitis C virus-specific cytotoxicity T lymphocyte epitopes / 中华微生物学和免疫学杂志

Objective To identify hepatitis C virus(HCV)-specific cytotoxicity T lymphocytes (CTL)epitopes by the combination of T epitopes prediction software and in vitro assays.Methods HCVspecific CTL epitopes were predicted by T epitope prediction software Rankpep and then candidate HCV-specific CTL epitopes were selected.Candidate HCV-specific CTL epitopes were used to stimulate PBMC of HCV-infected patients and healthy volunteers.and then enzyme-linked immunospot(ELISPOT)and intracellular cytokine staining(ICS)were used to measure the frequencies of IFN-γ-producing cells in total PBMC and the percentages of IFN-γ+CD8+T cells in total CD8+T cells,respectively.Results Five candidate CTL epitopes[NS3 450(TVPQDAVSR),NS3 594(GPTLLYRL),Ns4b 78(sMMAFSAAL),NS5a 416(SEENVSVVF)and NS5a 367(TVSSALAEL)]were used to stimulate PBMC of ten HCV-infected patients and two healthy volunteers.PBMC of seven HCV-infected patients secreted IFN-γ while PBMC of healthy volunteers did not produce IFN-γ.The frequencies of peptide-specific IFN-γ-producing cells ranged from 5 to 36 SFC/105 PBMC and the percentages of peptide-specific IFN-γ+CD8+T cells ranged from 0.02%-0.25%.Conclusion Resuhs of ELISPOT assay and ICS assay confirm that these five peptides NS3 450,NS3 594,NS4b 78,NS5a 416 and NS5a 367 are identified as Hovel HCV-specific CTL epitopes.

The HLA-A2 restriction and immunogenicity of hepatitis C virus-spedfic cytotoxic T lymphocyte epitopes / 中华微生物学和免疫学杂志

Objective To explore the HLA-A2 restriction and immunogenicity of 5 previously identified HCV-speeific CTL epitopes. Methods Based on T2 cell, to explore the HLA-A2 restriction of previously identified HCV-specific CTL epitopes by MHC-peptide complex stabilization assay;To detect pep-tide-specific CTL in HLA-A2~+ PBMC stimulated by HLA-A2-restricted peptides by intracellular cytokine staining(ICS) and ELISPOT; To explore the cytotoxicity of peptide-specific CTL to same peptide-loaded T2 cells (target cells) by CTL cytotoxicity test. Results Among 5 previously identified CTL epitopes NS4b_78 (SMMAFSAAL) and NS5a_367 (TVSSALAEL) have high-affinity for HLA-A2 molecules(FI 1) ;ELISPOT results shown that NS4b_78(SMMAFSAAL) and NSSa_367(TVSSALAEL) induced high levels of IFN-γ-se-creting cells [(60±6) SFC/10~4 PBMC vs (4±1 ) SFC/10~4 PBMC, P < 0.01 ; (10 ± 3 ) SFC/10~4 PBMC vs (2±1 ) SFC/10~4 PBMC, P <0.01, respectively] ;ICS results indicated that there were high percentages of CD8~+ IFN-γ~+ T cells in total CD8~+T cells stimulated by these peptides [(2.33 ±0.22 ) % vs (0.05±0.01)%, P <0.001 ; (0.36±0.06)% vs (0.03±0.01)%, P <0.001, respectively]. Furthermore,peptide-specific CTL could effectively kill same peptide-loadcd T2 cells. Conclusion NS4b_78 (SMMAF-SAAL) and NSSa_367 (TVSSALAEL) were identified as HLA-A2-restricted CTL epitopes which could in-duce immune response in vitro.

Effects of rosiglitazone on blood glucose, blood lipid and vascular remodeling in rats fed with high-fructose diet / 中国药理学通报

AIM To investigate the action of rosiglitazone on blood glucose,blood lipid,and the vascular remodeling in rats fed with high-fructose diet. METHODS After fed with high-fructose diet for one month,rats were randomly divided into two groups:High-fructose diet group that continued feeding with high-fructose diet for another month; High-fructose diet+rosiglitazone group fed with high-fructose diet and rosiglitazone (5 mg?kg -1?d -1, dissolved in water) for one month. At last,one half of each group were anesthetized,and the fasting blood, obtained from heart,was used to detect blood glucose,blood lipid,blood insulin. The aorta and mesenteric artery were gotten from the other half of each group, fixed in formalin,sliced and HE dyed. Pathology analysis system was used to measure the remodeling indexes of aorta and mesenteric artery:Lumen ,media,media/lumen,media cross-section area. RESULTS Comparing high-fructose diet+rosiglitazone group with high-fructose diet group, rosiglitazone decreased blood pressure(16.0 kPa vs 18.0 kPa, P

Immune response of peptide-specific CD4~+ T cells in mice immunized by Dengue virus-specific peptide immunization / 中国免疫学杂志

Objective:To explore the immunogenecity of four Dengue virus-specific peptides in mice.Methods:Each peptide of four Dengue virus-specific peptides (C45-57KLVMAFIAFLRFL,E396-408SSIGKMFEATARG,NS323-35YRILQRGLLGRSQ,and NS3141-155NREGKIVGLYGNGVV) was used to immunize BALB/c mice and C57BL/6 mice,respectively.Three weeks later,mice were killed and splelocytes were prepared.Splelocytes were stimulated by the same peptide and intracellular cytokine staining (ICS) assay was used to detect the percentages of peptide-specific IFN-? or IL-4 producing CD4+ T cells in total CD4+ T cells.Results:Peptide C45-57 induced peptide-specific IFN-?+ CD4+ T cells(0.72%?0.04% vs 0.04%?0.02%,P