Covariates of Cervical Cytokine mRNA Expression by Real-Time PCR in Adolescents and Young Women: Effects of Chlamydia trachomatis Infection, Hormonal Contraception, and Smoking
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Measurements of mucosal immune parameters in the uterine cervix are potentially influenced by numerous factors, including infections, endogenous and exogenous hormones, semen, and nicotine and its metabolites in cervical mucus. The objective of this study was to examine correlates of immunoregulatory cytokine mRNA expression in cervical cytology samples in a cross-sectional design. Samples, collected at study entry by cervical cytology brush from 368 women aged 13–21 enrolled in a longitudinal study of the natural history of human papillomavirus (HPV) infection, were tested by quantitative RT-PCR for expression of IFN-γ, IL-4, IL-10, and IL-12. In a multivariate analysis, elevated levels of IFN-γ, IL-10, and IL-12 were significantly (p < 0.05) associated with several variables, including current C. trachomatis infection, recent intercourse, and current oral contraceptive pill use. Suppressed IL-4 and IL-10 levels were associated with cigarette smoking within the last 24 h. Time since last menstrual period did not affect any of the cytokines; in a substudy of weekly cytokine variability, however, IL-10 showed a non-significant trend toward higher levels around the time of menstruation.
Key WordsAdolescent women uterine cervix biologic covariates cytokines mucosal immunity
This work was supported by PHS grants 2 R01 CA51323-11A1 and 2 R01 CA87905-01 and was carried out in part in the Pediatric Clinical Research Center, University of California, San Francisco, with funds provided by the National Center for Research Resources, 5 M01 RR-01271, U.S. Public Health Service. We would like to thank Roche Molecular Diagnostics for training and supplies to support the HPV DNA testing. We also thank Dr. David Ginzinger for his kind assistance in developing the quantitative RT‐PCR cytokine assays.
- 1.Schiffman MH, Bauer HM, Hoover RN, Glass AG, Cadell DM, Rush BB, Scott DR, Sherman ME, Kurman RJ, Wacholder S, Stanton CK, Manos MM: Epidemiologic evidence showing that human papillomavirus infection causes most cervical intraepithelial neoplasia. J Natl Cancer Inst 85(12):958–964, 1993PubMedCrossRefGoogle Scholar
- 2.Bosch FX, Manos MM, Muñoz N, Sherman M, Jansen AM, Peto J, Schiffman MH, Moreno V, Kurman R, Shah KV, International Biological Study on Cervical Cancer (IBSCC) Study Group: Prevalence of human papillomavirus in cervical cancer: A worldwide perspective [see comments]. J Natl Cancer Inst 87(11):796–802, 1995PubMedCrossRefGoogle Scholar
- 4.Moscicki AB, Hills N, Shiboski S, Powell K, Jay N, Hanson E, Miller S, Clayton L, Farhat S, Broering J, Darragh T, Palefsky J: Risks for incident human papillomavirus infection and low-grade squamous intraepithelial lesion development in young females. JAMA 285(23):2995–3002, 2001PubMedCrossRefGoogle Scholar
- 5.Moscicki AB, Shiboski S, Broering J, Powell K, Clayton L, Jay N, Darragh TM, Brescia R, Kanowitz S, Miller SB, Stone J, Hanson E, Palefsky J: The natural history of human papillomavirus infection as measured by repeated DNA testing in adolescent and young women. J Pediatr 132(2):277–284, 1998PubMedCrossRefGoogle Scholar
- 21.Ting Y, Manos MM: Detection and typing of genital human papillomavirus. In PCR Protocols: A Guide to Methods and Applications, M Innis, D Gelfand, J Sninsky, T White (eds). San Diego, California, Academic Press, 1990, pp 356–367Google Scholar
- 22.Molecular Research Center Inc.: TRI Reagent—RNA, DNA, protein isolation reagent. Manufacturer's protocol, 1995Google Scholar
- 27.Box GEP, Cox DR: An analysis of transformations. J R Stat Soc Ser B (Methodological) 26:211–246, 1964Google Scholar
- 30.Maret A, Coudert JD, Garidou L, Foucras G, Gourdy P, Krust A, Dupont S, Chambon P, Druet P, Bayard F, Guery JC: Estradiol enhances primary antigen-specific CD4 T cell responses and Th1 development in vivo. Essential role of estrogen receptor alpha expression in hematopoietic cells. Eur J Immunol 33(2):512–521, 2003PubMedCrossRefGoogle Scholar