Background Endocervical epithelial cells play early roles in the defense of upper female genital tract to pathogens. Toll-like receptors （TLRs） and human defensins （HD） have recently been identified as fundamental...Background Endocervical epithelial cells play early roles in the defense of upper female genital tract to pathogens. Toll-like receptors （TLRs） and human defensins （HD） have recently been identified as fundamental components of the innate immune responses to bacterial pathogens. We aimed to use in vitro model of human primary endocervical epithelial cells （HPECs） to investigate their roles in innate immune response of the endocervix. Methods TLR4 expression and distribution in HPECs and endocervix were investigated by immunofluorescence （IF）. Cultured HPECs were divided into lipopolysaccharide （LPS） group which were treated by LPS for 0, 24 and 48 hours, and control group without treatment. At each time point, the levels of HD5, IL-6 and TNF-a in supernants were determined by ELISA. TLR4 and HD5 expressions of cells were detected by Western blotting simultaneously. HD5 expression pattern was also compared between the HeLa cell line and HPECs. Results Endocervix tissue surface and HPECs expressed TLR4. After incubated with LPS, HPECs expressed significantly higher levels of TLR4 than control group, especially after 24 hours （P 〈0.01）, however decreased after 48 hours with a similar level of TLR4 expression compared with control group. LPS could upregulate the secretion of HD5, IL-6 and TNF-a in a time-dependent manner （24 hours： P 〈0.05; 48 hours： P 〈0.01, compared with control group）. Intracellular HD5 expression levels decreased over time. HD5 expression patterns in HPECs were different from HeLa cell line. Conclusions To respond to LPS stimulation, HPECs may function in the mucosal immune defense through TLR4 activation and HD5 secretion. HPEC is considered a significant model for immunological study.展开更多
文摘Background Endocervical epithelial cells play early roles in the defense of upper female genital tract to pathogens. Toll-like receptors （TLRs） and human defensins （HD） have recently been identified as fundamental components of the innate immune responses to bacterial pathogens. We aimed to use in vitro model of human primary endocervical epithelial cells （HPECs） to investigate their roles in innate immune response of the endocervix. Methods TLR4 expression and distribution in HPECs and endocervix were investigated by immunofluorescence （IF）. Cultured HPECs were divided into lipopolysaccharide （LPS） group which were treated by LPS for 0, 24 and 48 hours, and control group without treatment. At each time point, the levels of HD5, IL-6 and TNF-a in supernants were determined by ELISA. TLR4 and HD5 expressions of cells were detected by Western blotting simultaneously. HD5 expression pattern was also compared between the HeLa cell line and HPECs. Results Endocervix tissue surface and HPECs expressed TLR4. After incubated with LPS, HPECs expressed significantly higher levels of TLR4 than control group, especially after 24 hours （P 〈0.01）, however decreased after 48 hours with a similar level of TLR4 expression compared with control group. LPS could upregulate the secretion of HD5, IL-6 and TNF-a in a time-dependent manner （24 hours： P 〈0.05; 48 hours： P 〈0.01, compared with control group）. Intracellular HD5 expression levels decreased over time. HD5 expression patterns in HPECs were different from HeLa cell line. Conclusions To respond to LPS stimulation, HPECs may function in the mucosal immune defense through TLR4 activation and HD5 secretion. HPEC is considered a significant model for immunological study.
