J C Jeremias1, A M Bongiovanni, S S Witkin. 1. Department of Obstetrics and Gynecology, Weill Medical College of Cornell University, New York, NY 10021, USA. jaj2003@mail.med.cornell.edu
Abstract
OBJECTIVE: The 70kD heat shock protein (Hsp70), induced when cells are subjected to environmental stress, prevents the denaturation and incorrect folding of polypeptides and may expedite replication and transmission of DNA and RNA viruses. We analyzed whether messenger RNA (mRNA) for Hsp70 was expressed following exposure of a cultured human cervical cell line (HeLa cells) to human semen or in cervical cells from sexually active women. STUDY DESIGN: HeLa cells were co-cultured with a 1:50 dilution of semen from four men or with purified spermatozoa or cell-free seminal fluid. Endocervical swabs were acquired at mid-cycle from 53 women. Heat shock protein 70 mRNA was detected by a reverse transcriptase-polymerase chain reaction utilizing specific primer pairs and analysis on agarose gels. In cervical cells Hsp70 mRNA was measured identically followed by hybridization with an Hsp70-specific internal probe and detection by enzyme-linked immunosorbent assay (ELISA). Cervical immunoglobulin A (IgA) antibodies to the human Hsp70 were determined by ELISA. RESULTS: HeLa cell-semen co-culture resulted in the induction of Hsp70 mRNA. In addition, cell-free seminal plasma and motile sperm incubated individually with HeLa cells also induced this mRNA. Heat shock protein 70 mRNA was detected in 28 (52.8%) of 53 endocervical samples obtained from women at various time points following intercourse. The percentage of samples expressing this mRNA was 37.5% at less than 10 hours, 64.3% at 10 hours, 70% at 11 hours, and between 36% and 50% at later times after semen exposure. The detection of cervical IgA antibodies to the Hsp70 was highly associated with Hsp70 gene transcription. CONCLUSION: Human semen induces transcription of Hsp70 in cervical epithelial cells.
OBJECTIVE: The 70kD heat shock protein (Hsp70), induced when cells are subjected to environmental stress, prevents the denaturation and incorrect folding of polypeptides and may expedite replication and transmission of DNA and RNA viruses. We analyzed whether messenger RNA (mRNA) for Hsp70 was expressed following exposure of a cultured human cervical cell line (HeLa cells) to human semen or in cervical cells from sexually active women. STUDY DESIGN:HeLa cells were co-cultured with a 1:50 dilution of semen from four men or with purified spermatozoa or cell-free seminal fluid. Endocervical swabs were acquired at mid-cycle from 53 women. Heat shock protein 70 mRNA was detected by a reverse transcriptase-polymerase chain reaction utilizing specific primer pairs and analysis on agarose gels. In cervical cells Hsp70 mRNA was measured identically followed by hybridization with an Hsp70-specific internal probe and detection by enzyme-linked immunosorbent assay (ELISA). Cervical immunoglobulin A (IgA) antibodies to the humanHsp70 were determined by ELISA. RESULTS:HeLa cell-semen co-culture resulted in the induction of Hsp70 mRNA. In addition, cell-free seminal plasma and motile sperm incubated individually with HeLa cells also induced this mRNA. Heat shock protein 70 mRNA was detected in 28 (52.8%) of 53 endocervical samples obtained from women at various time points following intercourse. The percentage of samples expressing this mRNA was 37.5% at less than 10 hours, 64.3% at 10 hours, 70% at 11 hours, and between 36% and 50% at later times after semen exposure. The detection of cervical IgA antibodies to the Hsp70 was highly associated with Hsp70 gene transcription. CONCLUSION:Human semen induces transcription of Hsp70 in cervical epithelial cells.
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