AIM: To explore the role and significance of costimulatory molecules B7H1,B7H2 and ICOS within tissues of human gastric carcinoma and the possible mechanisms in tumor escape.
METHODS: mRNA expressions of costimulatory...AIM: To explore the role and significance of costimulatory molecules B7H1,B7H2 and ICOS within tissues of human gastric carcinoma and the possible mechanisms in tumor escape.
METHODS: mRNA expressions of costimulatory molecules including B7H1,B7H2,ICOS and B7-1 in tissues of human gastric carcinoma were investigated by in situ hybridization using digoxigenin-labeled oligonucleotide-probes. The tissue of chronic gastric ulcer was used as a control. All data were analyzed by SPSS statistic software.
RESULTS: At the site of gastric carcinoma, mRNA expression levels of B7H1, B7H2 and ICOS were much higher than that of B7-1. Their mRNA positive expression indexes were 0.512±0.333, 0.812±0.454, 0.702±0.359 and 0.293±0.253, respectively. The positively stained cells were mainly tumor infiltrating lymphocytes (TILs), and some tumor cells. The difference between them was greatly significant P＜0.005. The mRNA expression levels of four molecules were not correlated to the pathological grade and matastasis of gastric carcinoma.
CONCLUSION: ICOS-B7H costimulatory pathway may be predominant at the site of gastric carcinoma. B7-1mRNA might be the basis of ICOS-B7H interaction. ICOS-B7H interaction induces the production of IL-10 which inhibits the antitumor immune responses. Therefore, it is supposed that ICOS-B7H costimulatory pathway may be involved in the negative regulation of cell-mediated immune responses.展开更多
AIM： To observe the status of tumor-associated B7 molecule mRNA expression in human colorectal cancer tissue by/n situ hybridization. METHODS： The mRNA expression patterns of cancer- associated B7-1,B7H1t,B7H2, ICOS...AIM： To observe the status of tumor-associated B7 molecule mRNA expression in human colorectal cancer tissue by/n situ hybridization. METHODS： The mRNA expression patterns of cancer- associated B7-1,B7H1t,B7H2, ICOS in 22 specimens of human colorectal cancer tissue were monitored by in situ hybridization （ISH） with digoxin-labeled oligonucleotide probes. RESULTS： B7-1, B7H1,B7H2,ICOS mRNA were detected in both cancer cells and tumor infiltrating lymphocytes （TIL）. The mRNA expression level of these molecules in tumor cells was higher than that in TIL （0.76±0.54-1.62±0.82 vs 0.38±0.19-0.65±0.33, P〈0.001）. There was no relationship between expression level of tested B7 family molecules and patients＇ sex, age, differentiation status of cancer and regional lymph node metastasis. CONCLUSION： Th2 cytokine predominant in tumor microenvironment might be related to the expression of B7H1t B7H2 co-signal molecules in tumor cells and TIL. 2005 The WJG Press and Elsevier Inc. All rights reserved展开更多
To construct and express the fusion protein Stx2B-IntiminC300 of EHEC O157 ： H7, and to further investigate its immunoprophylactic potential, the gene of Stx2B （stx2b） from EHEC O157：H7 chromosome was cloned into ...To construct and express the fusion protein Stx2B-IntiminC300 of EHEC O157 ： H7, and to further investigate its immunoprophylactic potential, the gene of Stx2B （stx2b） from EHEC O157：H7 chromosome was cloned into pMD18-T vector. Thereafter, the amplified gene was cloned into prokary- otic expression plasmid pET-28a （ ＋ ）-eaeC300, which was constructed previously. The recombinant pasmid pET-28a（ ＋ ）-stx2b-eaeC300 was transformed into E. coli BL21 （DE3）. After inducement, the protein Stx2B-IntiminC300 was successfully expressed and analyzed with sodium dodecyl sulfate polyacrylamide gel electrophoresis （SDS-PAGE）, Western blotting and N-terminal amino acid residual sequencing. To evaluate its immunoprophylactic potential, it was primarily purified by ion-exchange chromatography and injected into 30 BALB/c mice with AI（OH）3 in the subscapular region. Ten days after the last booster vaccination, 20 mice were attacked with EHEC O157：H7 lysate and the protective efficacy was observed. In the present study, the gene of Stx2B-intiminC300 was successfully cloned into pET-28a （ ＋ ） vector. The results of SDS-PAGE and Western blotting assay showed that the fusion protein was successfully expressed in the inclusion body form, accounting for 25 % of total expression products, and its molecular weight was about 43 kDa. The result of the N-terminal amino acid residual sequencing showed that it was identical to that of the molecular designed. The purity was about 75 % after primary purification. Animal tests revealed that the fusion protein Stx2B-intiminC300 has elicited high titer of protective antibody relatively. These results demonstrate that the fusion protein Stx2B-IntiminC300 is successfully expressed in prokaryotic expression system and shows certain immunoprophylactic potential.展开更多
文摘AIM: To explore the role and significance of costimulatory molecules B7H1,B7H2 and ICOS within tissues of human gastric carcinoma and the possible mechanisms in tumor escape.
METHODS: mRNA expressions of costimulatory molecules including B7H1,B7H2,ICOS and B7-1 in tissues of human gastric carcinoma were investigated by in situ hybridization using digoxigenin-labeled oligonucleotide-probes. The tissue of chronic gastric ulcer was used as a control. All data were analyzed by SPSS statistic software.
RESULTS: At the site of gastric carcinoma, mRNA expression levels of B7H1, B7H2 and ICOS were much higher than that of B7-1. Their mRNA positive expression indexes were 0.512±0.333, 0.812±0.454, 0.702±0.359 and 0.293±0.253, respectively. The positively stained cells were mainly tumor infiltrating lymphocytes (TILs), and some tumor cells. The difference between them was greatly significant P＜0.005. The mRNA expression levels of four molecules were not correlated to the pathological grade and matastasis of gastric carcinoma.
CONCLUSION: ICOS-B7H costimulatory pathway may be predominant at the site of gastric carcinoma. B7-1mRNA might be the basis of ICOS-B7H interaction. ICOS-B7H interaction induces the production of IL-10 which inhibits the antitumor immune responses. Therefore, it is supposed that ICOS-B7H costimulatory pathway may be involved in the negative regulation of cell-mediated immune responses.
基金Supported by the National Natural Science Foundation of China, No. 30370549
文摘AIM： To observe the status of tumor-associated B7 molecule mRNA expression in human colorectal cancer tissue by/n situ hybridization. METHODS： The mRNA expression patterns of cancer- associated B7-1,B7H1t,B7H2, ICOS in 22 specimens of human colorectal cancer tissue were monitored by in situ hybridization （ISH） with digoxin-labeled oligonucleotide probes. RESULTS： B7-1, B7H1,B7H2,ICOS mRNA were detected in both cancer cells and tumor infiltrating lymphocytes （TIL）. The mRNA expression level of these molecules in tumor cells was higher than that in TIL （0.76±0.54-1.62±0.82 vs 0.38±0.19-0.65±0.33, P〈0.001）. There was no relationship between expression level of tested B7 family molecules and patients＇ sex, age, differentiation status of cancer and regional lymph node metastasis. CONCLUSION： Th2 cytokine predominant in tumor microenvironment might be related to the expression of B7H1t B7H2 co-signal molecules in tumor cells and TIL. 2005 The WJG Press and Elsevier Inc. All rights reserved
文摘To construct and express the fusion protein Stx2B-IntiminC300 of EHEC O157 ： H7, and to further investigate its immunoprophylactic potential, the gene of Stx2B （stx2b） from EHEC O157：H7 chromosome was cloned into pMD18-T vector. Thereafter, the amplified gene was cloned into prokary- otic expression plasmid pET-28a （ ＋ ）-eaeC300, which was constructed previously. The recombinant pasmid pET-28a（ ＋ ）-stx2b-eaeC300 was transformed into E. coli BL21 （DE3）. After inducement, the protein Stx2B-IntiminC300 was successfully expressed and analyzed with sodium dodecyl sulfate polyacrylamide gel electrophoresis （SDS-PAGE）, Western blotting and N-terminal amino acid residual sequencing. To evaluate its immunoprophylactic potential, it was primarily purified by ion-exchange chromatography and injected into 30 BALB/c mice with AI（OH）3 in the subscapular region. Ten days after the last booster vaccination, 20 mice were attacked with EHEC O157：H7 lysate and the protective efficacy was observed. In the present study, the gene of Stx2B-intiminC300 was successfully cloned into pET-28a （ ＋ ） vector. The results of SDS-PAGE and Western blotting assay showed that the fusion protein was successfully expressed in the inclusion body form, accounting for 25 % of total expression products, and its molecular weight was about 43 kDa. The result of the N-terminal amino acid residual sequencing showed that it was identical to that of the molecular designed. The purity was about 75 % after primary purification. Animal tests revealed that the fusion protein Stx2B-intiminC300 has elicited high titer of protective antibody relatively. These results demonstrate that the fusion protein Stx2B-IntiminC300 is successfully expressed in prokaryotic expression system and shows certain immunoprophylactic potential.
