CYTOKINES GENES PATTERN IN HUMAN PAPILLOMAVIRUS AND Chlamydia trachomatis CO-INFECTION AMONG WOMEN IN ILORIN, NIGERIA
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Date
2019-12
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UNIVERSITY OF ILORIN
Abstract
Human Papillomavirus (HPV) is a common sexually transmitted pathogen. HPV infection is a mandatory but not sufficient cause of cervical cancer. However, some women infected with HPV ultimately develop the disease. Several co-factors including Chlamydia trachomatis (CT) infection and pro-inflammatory cytokines are believed to influence the clinical outcome of HPV infection. Cytokines genes are polymorphic in nature and can lead to individual variation in cytokines production. This study assessed the cytokines genes pattern in HPV and CT co-infected women in Ilorin, Nigeria. The objectives were to: (i) identify participants with abnormal changes in cells of the cervix; (ii) detect HPV DNA in the exfoliated cervical cells of participants; (iii) determine the prevalence of HPV and the proportion of high risk HPV (HRHPV) among participants; (iv) identify CT DNA and its prevalence in cervical samples of participants; (v) determine the prevalence of HPV and CT co-infection; (vi) profile the cytokines genes associated with co-infection; and (vii) assess the risk factors associated with HPV and CT co- infections. Exfoliated cervical cells from 376 women were analysed by Papanicolaou test. Deoxyribonucleic acids (DNA) was extracted from the cells and amplified by Polymerase Chain Reaction (PCR) assays. MY09/11 and GP5+/6+ primers were used for amplification of HPV DNA by nested PCR; eighteen specific type primers for HRHPV DNA were amplified by nested multiplex PCR. Cryptic Plasmid primers were used to detect CT DNA by conventional PCR. Pro-inflammatory cytokines genes [Interferon gamma (IFN-γ) and Tumour Necrosis Factor alpha (TNF-α)] and anti-inflammatory cytokines genes [Interleukin-10 (IL-10) and Tumour Growth Factor-beta (TGF-β codons 10 and 25)] were amplified with Amplification Refractory Mutation System PCR. Statistical analysis of the data was done using chi-square, student t-test (t) and multiple regressions at p< 0.05. The findings of this study were that:
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(i) 19 (5.1%) women had abnormal cervical cytology out of which 13 (3.5%) had Low Squamous Intra-epithelial Lesion, 4 (1.1%) had Atypical Squamous Cell of Undetermined Significance and 2 (0.5%) had High Squamous Intra-epithelial Lesion; (ii) the fragments corresponded in position to base pairs (bp) of HPV (150 and 450 bp) and HRHPV (~118-457 bp); (iii) the prevalence of HPV was 81.4%, out of which 53.5% had HRHPV genotypes with HPV 82 being the most abundant (33.5%); (iv) CT fragments appeared at 201 bp, with a prevalence of 4.5%; (v) the prevalence of co-infection was 4.0%; (vi) comparison of co-infection with single infection showed significant differences (p< 0.05) in IFN-γ, TNF-α and TGF-β10. Also, when HPV and CT single infections were compared, IFN-γ and TNF-α showed significant difference (p< 0.05); (vii) HPV and CT individual infections were associated with risk factors such as age, parity, age at sexual debut, female genital cutting and contraceptive use. The study concluded that there were significant variations in predominantly pro-inflammatory cytokine genes in relation to HPV and CT co-infected women in Ilorin. The study recommended that these gene patterns may be used as predictor of HPV disease.
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Keywords
CYTOKINES, GENES PATTERN, HUMAN PAPILLOMAVIRUS, Chlamydia trachomatis, CO-INFECTION, WOMEN, ILORIN