RESUMEN
AIM: The aim of this study are to determine the hemodynamic changes in healthy patients during the surgical removal of lower third molar and to evaluate whether these variations are attributable to patient anxiety and pain experienced during surgical procedure. MATERIALS AND METHODS: Sixty healthy patients were evaluated (i) to determine the hemodynamic changes (systolic blood pressure [SBP], diastolic blood pressure [DBP], heart rate [HR], and oxygen saturation) at nine occasions: before starting the surgical procedure, 1 min and 4 min after local anesthetic injection, during the incision, at the time of ostectomy, at the completion of tooth removal, at the start and completion of suturing, and finally, after completion of surgery and (ii) to evaluate whether these variations are attributable to patient anxiety and pain experienced during the surgical procedure. Hemodynamic variables were compared between the gender and at different time points by performing two-way analysis of variance for repeated measures. Global mean values of hemodynamic variables were compared between male and female using unpaired t-test. Categorical variables were compared by Chi-square test. All the tests were two-sided. P < 0.05 was considered statistically significant. RESULTS: SBP and DBP showed significant changes; the highest value was recorded at the time of ostectomy/tooth sectioning. Maximum HR was observed 4 min after local anesthetic injection and the lowest HR was recorded after completion of tooth extraction, i.e., during the suturing. In females, mean HR was significantly increased. CONCLUSION: The present study suggests that dental anxiety impacts the effect of delivery of local anesthesia on blood pressure and is significantly associated with increased HR.
RESUMEN
PROBLEM: To study the innate immune response -TLR2 TLR 4 and iNOS expression in female genital Chlamydia trachomatis infection. METHOD: TLR 2, TLR 4, and iNOS expression was evaluated by real-time PCR in C. trachomatis-infected asymptomatic, mucopurulent cervicitis (MPC), and fertility disorders (FD) women. Expression of TLR signaling pathway genes was checked in vivo in C. trachomatis-infected cervical monocytes. Further, inos gene expression and nitric oxide release was assessed in vitro in THP-1 cell line upon chlamydial infection. RESULTS: TLR2, TLR4, and iNOS expression was significantly (P < 0.05) higher in C. trachomatis-positive women with FD, MPC, and asymptomatic women, respectively, than in control. Chlamydial infection significantly upregulates CD86, TLR4, MyD88, IRAK2, nF-κB, IL-1,ß and IL-12 genes. Expression of iNOS gene was found to be significantly (P < 0.05) high 12 hrs post-infection. CONCLUSIONS: Chlamydia trachomatis stimulates innate immune cells by activation of TLR2/TLR 4. Overall data indicate that recognition by TLR4 helps in initiation of immune response while recognition by TLR2 leads to secretion of inflammatory cytokines while iNOS-induced nitric oxide production helps in clearing Chlamydia. These results are first to provide initial insights into how innate immune response operates in human cervical monocytes upon chlamydial infection.
Asunto(s)
Cuello del Útero/inmunología , Chlamydia trachomatis/patogenicidad , Monocitos/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Receptor Toll-Like 2/metabolismo , Receptor Toll-Like 4/metabolismo , Adulto , Línea Celular , Cuello del Útero/microbiología , Infecciones por Chlamydia/inmunología , Infecciones por Chlamydia/microbiología , Femenino , Enfermedades de los Genitales Femeninos/inmunología , Enfermedades de los Genitales Femeninos/microbiología , Células HeLa , Humanos , Inmunidad Innata , Monocitos/inmunología , Óxido Nítrico Sintasa de Tipo II/genética , Receptor Toll-Like 2/genética , Receptor Toll-Like 4/genética , Regulación hacia ArribaRESUMEN
Chlamydiae are obligate intracellular bacteria that infect human epithelial cells. It has been reported that Chlamydia trachomatis, induces apoptosis in epithelial cells, however, the molecular mechanisms responsible for host cell death especially in primary epithelial cells remained largely unknown as most of the studies are in cell line like HeLa. In this study we demonstrated that C. trachomatis induces apoptosis signaling pathway and apoptosis in primary cervical epithelial cells in a time and dose dependent manner. Live cervical epithelial cells were isolated from endocervical cells and induction was done with chlamydial EBs. Our results demonstrated that apoptosis in infected epithelial cells was associated with an increased activity of caspase 8; however, caspase 9 was activated to a lesser extent. Analysis of apoptosis pathway revealed that expression level of McL-1, Bcl-2, CASP8, and TRADD genes were found to be significantly upregulated (P < 0.01), where as levels of Caspase 1, Caspase 10 and BRIC2 were found to be significantly downregulated (p < 0.01). Our results showed that Chlamydia induces apoptosis and caspase activation in epithelial cells through caspase 8, with an increased expression of the McL-1, which confers a block at the mitochondrial level.
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Caspasa 8/metabolismo , Infecciones por Chlamydia/inmunología , Chlamydia trachomatis/inmunología , Células Epiteliales/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Apoptosis/inmunología , Caspasa 8/genética , Técnicas de Cultivo de Célula , Células Cultivadas , Cuello del Útero/patología , Infecciones por Chlamydia/patología , Chlamydia trachomatis/patogenicidad , Activación Enzimática/inmunología , Células Epiteliales/inmunología , Células Epiteliales/patología , Femenino , Regulación de la Expresión Génica/inmunología , Humanos , Proteína 1 de la Secuencia de Leucemia de Células Mieloides , Proteínas Proto-Oncogénicas c-bcl-2/genética , Transducción de Señal/inmunologíaRESUMEN
Sexually transmitted Chlamydia trachomatis infection is an important public health concern with major adverse effects on female reproductive tract health and function. The magnitude of reproductive morbidity associated with sexually transmitted C. trachomatis infection is enormous, however to date no prophylactic vaccine is available. In part this is due to the lack of information on the mucosal immunobiology of the host-pathogen interaction and correlates of protective immunity during genital C. trachomatis infection. In this review, we focus on current knowledge of mucosal innate and adaptive immune responses in the female genital tract during C. trachomatis infection, which will eventually help in the development of a vaccine for prevention of chlamydial infection.
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Infecciones por Chlamydia/inmunología , Chlamydia trachomatis/inmunología , Genitales Femeninos/inmunología , Genitales Femeninos/microbiología , Inmunidad Mucosa , Enfermedades Bacterianas de Transmisión Sexual/inmunología , Vacunas Bacterianas , Infecciones por Chlamydia/prevención & control , Chlamydia trachomatis/patogenicidad , Femenino , Interacciones Huésped-Patógeno , Humanos , Infertilidad Femenina/etiología , Infertilidad Femenina/prevención & control , Enfermedades Bacterianas de Transmisión Sexual/prevención & controlRESUMEN
Chlamydia trachomatis is a leading cause of sexually transmitted infection worldwide and responsible for myriad of immunopathological changes associated with reproductive health. Delayed secretion of proinflammatory chemokine interleukin (IL)-8 is a hallmark of chlamydial infection and is dependent on chlamydial growth. We examined the effect of iron chelators on IL-8 production in HeLa 229 (cervix epitheloid cell, CCL2) cells infected with C. trachomatis. IL-8 production was induced by Iron chelator DFO and Mimosine, however, synergy with chlamydial infection was obtained with DFO only. Temporal expression of proinflammatory secreted cytokines IL-1beta, TNF-alpha, and IL-8 did not show synchrony in Chlamydia trachomatis infected cells. Secretion of IL-8 from Hela cells infected with C. trachomatis was not dependent on IL-1 beta and TNF- alpha induction. These results indicate towards involvement of iron in chlamydia induced IL-8 production.
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Infecciones por Chlamydia/metabolismo , Chlamydia trachomatis/crecimiento & desarrollo , Interleucina-8/metabolismo , Hierro/metabolismo , Ensayo de Inmunoadsorción Enzimática , Células HeLa , Humanos , Interleucina-1beta/metabolismo , Quelantes del Hierro/farmacología , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
The regulation of immune response and chlamydial infectious load in the cervix of human females is largely unknown. Infectious load in terms of inclusion-forming units (IFUs) was determined by quantitative cultures in Chlamydia-positive women, in asymptomatic women, women with mucopurulent cervicitis (MPC) and women with fertility disorders (FD). CD4(+), CD8(+), CD14(+) cells, myeloid and plasmacytoid dendritic cells (mDCs and pDCs) in the cervix were quantified by flow cytometry. Cervical cytokines, levels of beta-estradiol and C-reactive protein (CRP) in serum and cervical immunoglobulin A antibody to chlamydial major outer membrane protein antigen, chlamydial heat shock protein 60 and 10 antigens were measured by an enzyme-linked immunosorbent assay. In asymptomatic women, chlamydial load showed significant positive correlations with CD4, mDCs, interleukin-12 (IL-12) and IL-2; however, negative correlations were found with CD8 and IL-8 levels. In women with MPC, chlamydial IFUs correlated positively with CD8, pDC number, IL-8, CRP and interferon-gamma (IFN-gamma). In women with FD, chlamydial load showed a significant positive correlation with the pDC number, IL-10 and estradiol level and a negative correlation with CD4 and IFN-gamma. Overall, these results suggest that the interplay between chlamydial infectious load and host immune responses may be the deciding factor for the clinical condition presented during Chlamydia trachomatis infection.
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Cuello del Útero/patología , Infecciones por Chlamydia/inmunología , Chlamydia trachomatis/aislamiento & purificación , Recuento de Colonia Microbiana , Citocinas/análisis , Recuento de Leucocitos , Leucocitos/clasificación , Adulto , Anticuerpos Antibacterianos/análisis , Proteína C-Reactiva/análisis , Cuello del Útero/inmunología , Cuello del Útero/microbiología , Infecciones por Chlamydia/microbiología , Estradiol/análogos & derivados , Estradiol/sangre , Femenino , Humanos , Inmunoglobulina A/análisis , Leucocitos/inmunología , Adulto JovenRESUMEN
BACKGROUND: Chlamydia trachomatis infection of the female genital tract can lead to serious sequelae resulting in fertility related disorders. Little is known about the mechanism leading to Chlamydia induced pathology and factors responsible for it. As only some of the women develops reproductive disorders while majority of the women clears infection without any severe sequalae, mucosal immune response in women with or without fertility disorders was studied to identify factors which may lead to final clinical outcome of chlamydial infection. METHODS: Myeloid DCs (mDCs) and plasmacytoid DCs (pDCs) populations in cervical mucosa and peripheral blood were analyzed in controls and Chlamydia positive women with or without fertility disorders with multicoloured flow cytometric analysis. Cervical cytokines (IL-6, IL-8, IL-10, IL-12, TNF-alpha and IFN-gamma), C-reactive protein levels and sex hormone levels in serum were quantified by ELISA. RESULTS: In cervix of Chlamydia positive women with fertility disorders, significantly high (P < 0.05) numbers of pDCs were present with increased CD80 expression. pDCs correlated significantly with C-reactive protein levels, IL-6 and IFN-gamma levels in women with fertility disorders. In contrast, mDCs showed significant upregulation of CD1a during chlamydial infection and correlated significantly with IL-12 levels in Chlamydia positive fertile women. beta-estradiol levels were significantly higher in women having fertility disorders as compared to fertile women and have significant correlations (r = 0.65; P < 0.05) with pDCs numbers, CD80 expression, IL-6 levels and IFN-gamma levels in these women. CONCLUSION: These results suggest that development of sequalae in some women can be a result of interplay of many factors including type of dendritic cell, co stimulatory molecule expression, cytokine secretion pattern and hormone levels.
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Cuello del Útero/inmunología , Infecciones por Chlamydia/complicaciones , Citocinas/fisiología , Células Dendríticas/fisiología , Estradiol/fisiología , Adulto , Antígenos de Superficie/metabolismo , Proteína C-Reactiva/metabolismo , Estudios de Casos y Controles , Cuello del Útero/patología , Infecciones por Chlamydia/sangre , Infecciones por Chlamydia/inmunología , Infecciones por Chlamydia/patología , Chlamydia trachomatis/inmunología , Citocinas/metabolismo , Células Dendríticas/metabolismo , Células Dendríticas/patología , Estradiol/sangre , Femenino , Fertilidad/inmunología , Enfermedades de los Genitales Femeninos/sangre , Enfermedades de los Genitales Femeninos/etiología , Enfermedades de los Genitales Femeninos/inmunología , Enfermedades de los Genitales Femeninos/microbiología , Humanos , Inmunidad Mucosa/inmunología , Embarazo , Complicaciones Infecciosas del Embarazo/sangre , Complicaciones Infecciosas del Embarazo/etiología , Complicaciones Infecciosas del Embarazo/inmunología , Complicaciones Infecciosas del Embarazo/microbiología , Progesterona/sangreRESUMEN
Chlamydia trachomatis infection is followed by the development of antigen-specific cell-mediated immunity, which is detectable as a positive lymphocyte proliferation response to the chlamydial major outer membrane protein (MOMP) antigen. To date, however, there have been no studies on the mucosal immune responses to chlamydial antigens. This study aimed to study the primary and secondary immune responses of cervical lymphocytes in response to the chlamydial antigen. Median proliferative responses were found to be significantly (P<0.05) higher in patients with chlamydial infections than in controls. The chlamydial MOMP induced significantly higher IL-6 and IL-10 and lower interferon-gamma (IFN-gamma) secretion in cervical lymphocytes of Chlamydia-positive women, resulting in a T helper 2 response. On stimulation of peripheral blood mononuclear cells (PBMC) obtained from Chlamydia-positive women with the chlamydial antigen, the median levels of IL-10, IL-12 and IFN-gamma were higher than in controls, but the differences were not significant. Our study suggests that the mucosal immune responses towards Chlamydia trachomatis are different from those of PBMCs and are more helpful in understanding the cytokine responses in the female genital tract during chlamydial infection.
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Cuello del Útero/inmunología , Infecciones por Chlamydia/inmunología , Chlamydia trachomatis/inmunología , Inmunidad Mucosa , Activación de Linfocitos , Linfocitos/inmunología , Proteínas de la Membrana Bacteriana Externa/inmunología , Proliferación Celular , Femenino , Humanos , Interferón gamma/biosíntesis , Interleucina-10/biosíntesis , Interleucina-6/biosíntesis , Leucocitos Mononucleares/inmunologíaRESUMEN
Little is known about concurrent expression of cervical cytokines and their regulation by sex hormones during primary or recurrent chlamydial infections in humans. Cytokine (interleukin-1beta [IL-1beta], IL-6, IL-10, interferon-gamma [IFN-gamma], and tumor necrosis factor-alpha [TNF-alpha]) concentrations in cervical washes and serum samples, along with levels of beta-estradiol and progesterone in women with primary or recurrent chlamydial infections and healthy controls, were measured by ELISA. Women with recurrent infections had significantly higher levels of IFN-gamma in cervical washes than did women with primary infections. Significant negative correlation was found between IL-1beta and progesterone levels during recurrent infections. Beta-estradiol levels in women with primary infections showed significant negative correlations with cervical concentrations of IL-10, IL-1beta, and IL-6. Our study suggests that Chlamydia trachomatis infection in the female genital tract may be regulated by both the synergistic actions of the cytokines and the sex hormones beta-estradiol and progesterone.
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Cuello del Útero/inmunología , Infecciones por Chlamydia/inmunología , Chlamydia trachomatis/inmunología , Citocinas/inmunología , Adulto , Cuello del Útero/metabolismo , Cuello del Útero/microbiología , Infecciones por Chlamydia/sangre , Infecciones por Chlamydia/microbiología , Citocinas/sangre , Estradiol/sangre , Estradiol/inmunología , Femenino , Humanos , Persona de Mediana Edad , Progesterona/sangre , Progesterona/inmunologíaRESUMEN
PROBLEM: Chlamydial infections are often associated with various fertility-related disorders. Serological prediction of these has limitations, as they do not differentiate between past and current infections. Thus, we looked for local markers that could predict more precisely women at higher risk of developing severe complications. METHOD OF STUDY: A total of 320 Chlamydia trachomatis positive women with or without fertility disorders were tested for the prevalence of immunoglobulin A antibodies to synthetic peptides of chlamydial heat-shock protein 60 (cHSP60) and cHSP10 along with cervical interferon-gamma (IFN-gamma) and serum C-reactive protein (CRP) levels. RESULTS: Positive IFN-gamma level was the single best predictor for fertility disorder [odds ratio (OR) 15.4]. The predictive value of IFN-gamma could be significantly improved only by the addition of CRP test (OR 37.9). CONCLUSION: Positive IFN-gamma levels in cervical washes along with elevated CRP levels could be used to predict women who are at higher risk of developing fertility disorders.