Background The most critical mechanism governing drug resistance in Candida albicans （C. albicans） involves efflux pumps, the functionality of which largely depends on energy metabolism. Alcohol dehydrogenase I （A...Background The most critical mechanism governing drug resistance in Candida albicans （C. albicans） involves efflux pumps, the functionality of which largely depends on energy metabolism. Alcohol dehydrogenase I （ADH1） plays an important role in intracellular energy metabolism. The aim of this study was to explore the relationship between ADH1 and drug resistance in C. albicans. Methods Twenty clinical C. albicans samples isolated from individual patients diagnosed with vulvovaginal candidiasis, and two C. albicans strains obtained from a single parental source （the fluconazole （FLC）-sensitive strain CA-1s and the FLC-resistant strain CA-16R） were included in our study. In accordance with the Clinical and Laboratory Standards Institute （CLSI） M27-A3 guidelines, we used the microdilution method to examine the FLC minimum inhibitory concentrations （MICs） and real-time reverse transcription polymerase chain reaction （RT-PCR） to measure the mRNA expression levels of ADH1 and the azole resistance genes CDR1, CDR2, MDR1, FLU1 and ERG11 in all the isolates. Results A highly significant positive correlation between the mRNA levels of ADH1 and the MICs （r =0.921, P=0.000）, as well as positive correlations between the mRNA level of ADH1 and those of CDR1, CDR2 and FLU1 （rs of 0.704, 0.772 and 0.779, respectively, P 〈0.01）, were observed in the 20 clinical C. albicans samples. The relative expression of ADH1 was upregulated 10.63- to 17.61-fold in all of the drug-resistant isolates. No correlations were found between the mRNA levels of ADH1 and those of MDR1 or ERG11 （P 〉0.05）. The mRNA levels of the examined drug resistance genes were higher in the CA-16R strain than in CA-1s, and the mRNA levels of ADH1 in CA-16R were 11.64-fold higher than those in CA-1s （P 〈0.05）. Conclusions These results suggest that high levels of ADH1 transcription are implicated in FLC resistance in C. albicans and that the mRNA expression levels of ADH1 are positively correlated with those of CDR1, CDR2 and FLU1.展开更多
Adoptive cell-transfer therapy （ACT） has been reported to suppress growing tumors and to overcome tumor escape in animal models. As a candidate ACT effector, γ9δ2T cells can be activated and expanded in vitroand i...Adoptive cell-transfer therapy （ACT） has been reported to suppress growing tumors and to overcome tumor escape in animal models. As a candidate ACT effector, γ9δ2T cells can be activated and expanded in vitroand in vivoand display strong antitumor activity against colorectal, lung, prostate, ovarian and renal cell carcinomas. However, it is difficult to obtain a large enough number of γδT cells to meet the need for immunotherapy that can overcome the cancer patients＇ immune suppressive tumor microenvironment. In previous studies, our lab confirmed that γ9δ2T cells recognized tumor cells via the CDR3δ region of the γδ-T-cell receptor （TCR）. We constructed full-length human peripheral blood mononuclear cell （PBMC）-derived γ9 and δ2 chains in which the CDR3 region was replaced by an ovarian epithelial carcinoma （OEC）-derived CDR3. We transferred the CDR3δ-grafted γ9δ2TCR into peripheral blood lymphocytes （PBLs） to develop genetically modified γ9δ2T cells. In vitro studies have shown that these CDR3δ-grafted γ9δ2T cells can produce cytokines after stimulation with tumor cell extracts and exhibit cytotoxicity towards tumor cells, including human OEC and cervical adenocarcinoma. CDR3δ-grafted γ9δ2T cells adoptively transferred into nude mice bearing a human OEC cell line demonstrated significant antitumor effects. These results indicate that CDR3δ-grafted γ9δ2T cells might be candidates for clinical tumor immunotherapy.展开更多
基金This study was supported by grants from the National Natural Science Foundation of China （Nos. 81171542, 30972660）, the Natural Science Foundation of Guangdong Province, China （No. 10151008901000131）, Guangzhou Key Technology RD Program, China （No. 2010J-E011）, and the Fundamental Research Funds for the Central Universities （No. 21611509）.
文摘Background The most critical mechanism governing drug resistance in Candida albicans （C. albicans） involves efflux pumps, the functionality of which largely depends on energy metabolism. Alcohol dehydrogenase I （ADH1） plays an important role in intracellular energy metabolism. The aim of this study was to explore the relationship between ADH1 and drug resistance in C. albicans. Methods Twenty clinical C. albicans samples isolated from individual patients diagnosed with vulvovaginal candidiasis, and two C. albicans strains obtained from a single parental source （the fluconazole （FLC）-sensitive strain CA-1s and the FLC-resistant strain CA-16R） were included in our study. In accordance with the Clinical and Laboratory Standards Institute （CLSI） M27-A3 guidelines, we used the microdilution method to examine the FLC minimum inhibitory concentrations （MICs） and real-time reverse transcription polymerase chain reaction （RT-PCR） to measure the mRNA expression levels of ADH1 and the azole resistance genes CDR1, CDR2, MDR1, FLU1 and ERG11 in all the isolates. Results A highly significant positive correlation between the mRNA levels of ADH1 and the MICs （r =0.921, P=0.000）, as well as positive correlations between the mRNA level of ADH1 and those of CDR1, CDR2 and FLU1 （rs of 0.704, 0.772 and 0.779, respectively, P 〈0.01）, were observed in the 20 clinical C. albicans samples. The relative expression of ADH1 was upregulated 10.63- to 17.61-fold in all of the drug-resistant isolates. No correlations were found between the mRNA levels of ADH1 and those of MDR1 or ERG11 （P 〉0.05）. The mRNA levels of the examined drug resistance genes were higher in the CA-16R strain than in CA-1s, and the mRNA levels of ADH1 in CA-16R were 11.64-fold higher than those in CA-1s （P 〈0.05）. Conclusions These results suggest that high levels of ADH1 transcription are implicated in FLC resistance in C. albicans and that the mRNA expression levels of ADH1 are positively correlated with those of CDR1, CDR2 and FLU1.
文摘Adoptive cell-transfer therapy （ACT） has been reported to suppress growing tumors and to overcome tumor escape in animal models. As a candidate ACT effector, γ9δ2T cells can be activated and expanded in vitroand in vivoand display strong antitumor activity against colorectal, lung, prostate, ovarian and renal cell carcinomas. However, it is difficult to obtain a large enough number of γδT cells to meet the need for immunotherapy that can overcome the cancer patients＇ immune suppressive tumor microenvironment. In previous studies, our lab confirmed that γ9δ2T cells recognized tumor cells via the CDR3δ region of the γδ-T-cell receptor （TCR）. We constructed full-length human peripheral blood mononuclear cell （PBMC）-derived γ9 and δ2 chains in which the CDR3 region was replaced by an ovarian epithelial carcinoma （OEC）-derived CDR3. We transferred the CDR3δ-grafted γ9δ2TCR into peripheral blood lymphocytes （PBLs） to develop genetically modified γ9δ2T cells. In vitro studies have shown that these CDR3δ-grafted γ9δ2T cells can produce cytokines after stimulation with tumor cell extracts and exhibit cytotoxicity towards tumor cells, including human OEC and cervical adenocarcinoma. CDR3δ-grafted γ9δ2T cells adoptively transferred into nude mice bearing a human OEC cell line demonstrated significant antitumor effects. These results indicate that CDR3δ-grafted γ9δ2T cells might be candidates for clinical tumor immunotherapy.
