Systemic and mucosal concentrations of nine cytokines among individuals with Neisseria gonorrhoeae infection in Nairobi, Kenya
Description
We conducted a cross-sectional comparative study of consecutive adults, 18 to 49 years, seeking treatment for sexually transmitted infections at Casino Special Treatment Centre (STC) Health Centre in Nairobi, Kenya. Patients were included if they were between 18 and 49 years and provided written informed consent to participate in the study. An interviewer-administered questionnaire was used to collect the sociodemographic characteristics and the sexual history. Vaginal samples were collected after insertion of a speculum. For detection of Neisseria gonorrhoeae infection, a dacron swab was placed approximately 2cm into the os and rotated three times, removed and placed into an empty tube and immediately placed on ice. Cervicovaginal lavage (CVL) was collected for mucosal cytokines. Briefly, the cervix and the lateral vaginal walls were flushed with 10 ml normal saline at room temperature. The fluid was allowed to pool into the posterior fornix and aspirated into the same syringe. This procedure was repeated 3-5 times with the same fluid. The fluid was then collected and placed in a 15ml falcon tube and immediately placed on ice to await transportation, up to 4 hours. Urethral samples were collected by gently inserting dacron swabs 2-4 cm inside the urethra and rotating three times. They were then carefully removed and placed (i) into an empty tube for detection of Neisseria gonorrhoeae infection and (ii) in a tube containing 1.5ml phosphate buffered saline (PBS) for mucosal cytokine analysis. The tubes were immediately placed on ice to await transportation. For systemic cytokine analysis, 10mls of venous blood was collected, from all participants, into EDTA treated tubes and then immediately placed on ice to await transportation. All participants were tested for HIV using Determine HIV Rapid Test (Abbott Diagnostics) with confirmatory testing being done with the First Response™ (Premier Medical Corporation Private Ltd., Gujarat, India), according to Kenya’s Ministry of Heath guidelines. Those who tested negative with the first test were considered HIV negative while those who tested positive with the confirmatory test were considered HIV positive and were either enrolled in the Casino H/C HIV comprehensive care clinic (CCC) or referred to a health facility of their choice for follow-up. Screening for Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis and Mycoplasma genitalium was done using Multiplex PCR (Sacace Biotechnologies, Como, Italy) according to the manufacturer’s instructions. The systemic and mucosal levels of 9 cytokines (IL-1, IL-2, IL-4, IL-6, IL-10, IL-12p70, IL-17A, TNF- and IFN-) were measured using the Invitrogen™ High Sensitivity 9-Plex Human ProcartaPlex™ Panel (Thermo Fisher Scientific Inc.)