ABSTRACT
Genital Chlamydia trachomatis infection creates a substantial reproductive health burden in women. The high incidence of asymptomatic infection often precludes timely antibiotic therapy to control the sequelae of infection, and therefore a vaccine is required. Dendritic cells (DC) are now being used as an adjuvant for vaccine development; however, the fate of C. trachomatis in human DC and differential regulation of cytokine secretion remains unclear. Hence, an in vitro study was performed using C. trachomatis (serovar D) elementary body (EB)-pulsed, monocytederived DCs co-cultured with autologous CD4+ T cells. Secreted cytokines were measured to assess the protective/pathogenic immune response. The effect of (beta-oestradiol in the modulation of DC function and on Toll-like receptor (TLR) gene expression was also studied. Elementary body-pulsed DCs showed induction of protective Th1 immune response with upregulation of TLR4 expression, secretion of interleukin (IL)-6, IL-12 and interferon (IFN)-y, together with upregulation of major histocompatibility complex (MHC) class II, CD83 and CD86. When co-cultured with autologous CD4+T cells, DCs presented chlamydial antigens efficiently, as shown by proliferation of T cells and secretion of IL-2 and IFN gamma, which provide a protective immune response. However; pretreatment of cells with oestradiol significantly reduced TLR4 expression and upregulated IL-10 secretion, modulating the Th1 immune response to a Th2-type response, which may lead to pathogenesis.
Subject(s)
CD4-Positive T-Lymphocytes/drug effects , Chlamydia Infections/immunology , Chlamydia trachomatis/immunology , Estradiol/pharmacology , Macrophages/drug effects , Toll-Like Receptor 4/immunology , CD4-Positive T-Lymphocytes/immunology , CD4-Positive T-Lymphocytes/microbiology , Coculture Techniques , Epithelial Cells/cytology , Epithelial Cells/immunology , Epithelial Cells/microbiology , Estrogens/pharmacology , Female , HeLa Cells , Humans , Macrophages/immunology , Macrophages/microbiology , Signal Transduction/immunologySubject(s)
Anti-Infective Agents/administration & dosage , Chlamydia Infections/drug therapy , Plant Preparations/administration & dosage , Administration, Intravaginal , Aloe , Chlamydia trachomatis , Curcumin , Drug Combinations , Drug Evaluation , Female , Humans , Ointments , Phytotherapy/methods , Rosa , TabletsABSTRACT
Little is known about genital mucosal immune response to chlamydial infection in women with or without sequelae (Chlamydia positive women with or without fertility disorders as infertility and multiple spontaneous abortions). Cervical lymphocytes were stimulated with chlamydial EBs and cytokine secretion was determined by ELISA, RT-PCR and ELISPOT assays. Stimulated cervical cells from women with fertility disorders (FD) secrete significantly (P<0.05) higher levels of IL-1beta, IL-6, IL-8 and IL-10 and cells from fertile women secrete significantly higher levels of IL-12 and IFN-gamma compared to other groups. RT-PCR analysis showed similar results for IFN-gamma and IL-12. For IL-10 and IL-4, mRNA expression levels were significantly higher (P<0.05) in cells obtained from women with FD compared to other groups. Results for ELISPOT assay were similar as those of RT-PCR. The results suggest that cytokine secretion profile of cervical cells may decide whether infection does not hamper fertility or will develop fertility disorder.
Subject(s)
Cervix Uteri/immunology , Chlamydia Infections/immunology , Cytokines/immunology , Cytokines/metabolism , Genital Diseases, Female/immunology , Mucous Membrane/immunology , Adult , Cells, Cultured , Cervix Uteri/cytology , Chlamydia Infections/epidemiology , Chlamydia Infections/prevention & control , Chlamydia trachomatis/immunology , Cytokines/genetics , Enzyme-Linked Immunosorbent Assay , Female , Flow Cytometry , Genital Diseases, Female/pathology , Humans , India/epidemiology , Mucous Membrane/cytology , RNA, Messenger/metabolism , Reference Standards , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
The effect of a novel polyherbal formulation BASANT on Chlamydia trachomatis was studied. In vitro sensitivity testing was done by direct exposure of C. trachomatis (pre-infection incubation with BASANT) and exposure of C. trachomatis within HeLa 229 cells (post-infection incubation with BASANT). Pre-infection incubation of standard serovar D/UW-3/Cx with BASANT showed complete inhibition after 60, 30 and 15 min of incubation at concentrations of 12, 30 and 60 microg/mL, respectively. In the post-infection incubation, 8-10 microg/mL of BASANT showed complete inhibition of standard serovar D/UW-3/Cx as well as of five clinical isolates of C. trachomatis after 48 h of incubation. BASANT also inhibited a clinical isolate obtained from a doxycycline treatment failure patient at a concentration of 30 microg/mL. Both assays with standard and clinical isolates showed that BASANT has antimicrobial activity against C. trachomatis, suggesting the potential clinical utility of BASANT for the prevention of C. trachomatis infection by the sexual route.
