Expression Of TNF-α Of The PBMC Induced By Candida Albicans Insoluble β-glucan

Objective:Candida albicans insolubleβ-glucan(CAIBG)is a kind of biological response modifiers(BRM)which has extensive biological activities,such as increasing the number of leukocytes,anti-tumor activity,anti-virus activity and so on. Previous experiments of our research team have successfully isolated CAIBG from the cell walls of Candida albicans,and confirmed that CAIBG is able to inhibit the immuno-suppression effect of CTX in animal experiments,meanwhile CAIBG can elevate the number of leukocyte of peripheral blood in mice and can induce the introduction of TNF-αin blood.In this present study,we extracted Candida albican insolubleβ-glucan from cultured Candida albicans,and CAIBG on human peripheral blood mononuclear cells(PBMC)activity and TNF-αexpression were detected,and we also study the mechanism of its inducing cytokines to find the relationship between cytokine and the effect of anti-tumor.Method:Firstly we extracted insolubleβ-glucan from Candida albicans.PBMC was cultured in the medium by adding different concentrations of CAIBG(31.25ug/ml, 62.5ug/ml,125ug/ml,250ug/ml,and 500ug/ml).After 12 hours,24 hours and 36 hours respectively,we detected the activity of the PBMC,meanwhile join the CAIBG after 3 hours,6hours,12hours,18hours and 24hours to detect the TNF-αlevels for recognition the peak point time and the recession time,then in the peak point time,by adding different concentrations of CAIBG(31.25ug/ml,62.5ug/ml,125ug/ml, 250ug/ml,500ug/ml),we detected TNF-αlevels for recognition of the drug dose-effect relationship.Human pulmonary adenocarcinoma cells were cultured,the effect of PBMC cell line as effective cells,human pulmonary adenocarcinoma cell line as target cells,designed 5:1,10:1,20:1 three target efficiency ratio,were randomly divided into four groups:the control group,negative control group,the positive control group,and CAIBG groups.The cytotoxicity of the PBMC to the lung adenoearcinoma cells was detected,and also TNF-αlevel was examined. Results:Compared with the control group,CAIBG concentration of 62.5ug/ml and 125ug/ml for 12 hours,24 hours and 36 hours,the activity of the PBMC was significantly increased(P＜0.05).CAIBG on PBMC can effectively promote the secretion of TNF-αin all groups after joining of drugs and a clear time-dependent manner,but on 18hours,there was statistic significance between experiment and the control group(P＜0.05).Given different concentrations of the drug after 18 hours,the secretion of TNF-αhad significantly dose-dependent manner,but joining concentration of 62.5ug/ml,there was statistic significance between experiment and the control group(P＜0.05).However the effective cells:target cells was 5:1,10:1 or a ratio of 20:1,giving drug CAIBG concentration of 62.5ug/ml,PBMC toxic effects compared with untreated normal control group,negative control group,there was a significant difference(P＜0.05),while CAIBG group and cisplatin group had no statistically significant difference(P＞0.05);The secretion of TNF-αcompared with untreated normal control group,negative control group and cisplatin group,there was a significant difference(P＜0.05).Conclusion:This study suggested that,CAIBG may induce TNF-αsecretion of the PBMC,thereby killing tumor cells,which may be one of the mechanisms of anti-tumor induced by CAIBG.CAIBG is a kind of promising biological response modifier which is worthy of further research and development.