A distinct cellular profile is seen in the human endocervix during Chlamydia trachomatis infection

Problem The endocervix is a major target of Chlamydia trachomatis infection, but little is known about the immune repertoire in this tissue, or its response to these common bacteria. Method of study Using a cytobrush, we isolated cells from the endocervix of 20 women during C. trachomatis infection, and post‐antibiotic treatment. Endocervical swabs and blood were taken in parallel. Endocervical cells were enumerated, and endocervical and blood T cells immunophenotyped. Chlamydia trachomatis was genotyped by sequence analysis of the OmpA gene, and quantified by culture. Results Chlamydia trachomatis genotypes were D, E, F and Ia, and infectious burden varied considerably. Endocervical T cell and neutrophil numbers were highly elevated during infection, with both CD4 and CD8 T‐cell subsets accumulating. Regardless of the presence or absence of infection, the endocervical cell infiltrate was dominated by effector memory T cells, and the numbers of CCR5 and CD103 expressing T cells was significantly higher than in the blood. Human leukocyte antigen (HLA‐DR) expression by endocervical T cells was significantly increased during infection. Conclusion The human endocervix exhibits a distinct cellular response to C. trachomatis infection that can be longitudinally evaluated by cytobrush sampling. Infecting organisms can be sampled and analyzed in parallel.     

Intravaginal inoculation of mice with the Chlamydia trachomatis mouse pneumonitis biovar results in infertility.

In an attempt to establish a model of chlamydial ascending salpingitis and infertility, three inbred strains of mice, C3H/HeN (H-2k), C57BL/6N (H-2b), and BALB/cAnN (H-2d), were inoculated intravaginally with 3 x 10(7) inclusion-forming units of the Chlamydia trachomatis mouse pneumonitis biovar. Mice mated 6 weeks following inoculation were found to have a significant decrease in fertility rate compared with the control groups, as shown by a reduction in the number of pregnant mice and a decrease in the number of embryos.     

Chemokine bioactivity of RANTES is elevated in the sera of infertile women with past Chlamydia trachomatis infection

PROBLEM: It has been shown that Chlamydia trachomatis infection in infertile women is highly associated with tubal pathology. Chlamydia trachomatis antibody testing is a simple screening test for tubal factor subfertility, however, it is based on the detection of previous infection. Recently, association between some inflammatory diseases and chemokines has been investigated. This study was performed to clarify the relationship between chemokines in the sera of infertile women and past C. trachomatis infection. METHOD OF STUDY: Serum samples were collected from 10 infertile women having C. trachomatis antibodies [immunoglobulin (Ig)G and/or IgA] in their sera and 10 infertile women without the antibodies. All patients' tubo-ovarian structures were explored by transvaginal hydrolaparoscopy (THL). A CXC chemokine, interleukin-8, and six CC chemokines including macrophage inflammatory protein-1alpha (MIP-1alpha), MIP-1beta, monocyte chemotactic protein-1 (MCP-1), MCP-3, eotaxin, and regulated on activation, normal T cell expressed and secreted (RANTES) concentrations in their sera were analyzed using enzyme-linked immunosorbent assay. RESULTS: The serum concentration of RANTES was significantly higher in patients with C. trachomatis antibodies than those without the antibodies (P = 0.019). However, there were no significant differences of the concentrations of other chemokines between the sera of infertile women with and without C. trachomatis antibodies. The concentration of RANTES in the sera of infertile women did not correlate with C. trachomatis antibody titers or tubal pathology diagnosed by THL. CONCLUSIONS: These findings might indicate significant roles of some chemokines in the pathogenesis of C. trachomatis infection. Further studies are necessary to study the usefulness of evaluations of chemokines in tubal infertility investigations.     

密码修饰可增强沙眼衣原体MOMP DNA质粒免疫小鼠的免疫应答

目的 探讨密码修饰对沙眼衣原体主要外膜蛋白（MOMP）基因表达和DNA质粒免疫的影响。方法 根据人类密码子使用偏好对鼠肺炎沙眼衣原体（Chlamydia trochomatis mouse pneumonitis）MOMP基因进行优化修饰，设计合成人源化MOMP基因（HuMOMP）。构建以pcDNA3为载体的含HuMOMP及含野生型MOMP基因（WtMOMP）的真核表达质粒。瞬时转染COS-1细胞，Western blot方法比较HuMOMP基因及WtMOMP基因在哺乳动物细胞中的蛋白表达水平。DNA质粒肌内免疫BALB／c小鼠，检测小鼠血清特异性抗体、DTH和淋巴细胞增殖反应，比较优化密码MOMP基因和野生型MOMP基因DNA质粒免疫的免疫效果。结果 人工合成HuMOMP基因，成功构建重组真核表达质粒pcDNA3-HuMOMP及pcDNA3-WtMOMP。转染细胞Western blot结果显示，HuMOMP基因的蛋白表达水平明显高于WtMOMP基因。ELISA结果表明，HuMOMP DNA质粒免疫组小鼠血清特异性IgG抗体有所升高；细胞免疫检测结果显示，HuMOMP DNA质粒免疫组小鼠足垫肿胀程度增强、淋巴细胞增殖实验刺激指数（SI）增高，两者与WtMOMP DNA质粒免疫组相比P〈0．05，差异均有统计学意义。结论 人源化密码修饰能够增强沙眼衣原体MOMP基因在哺乳动物细胞中的蛋白表达及DNA质粒免疫小鼠的免疫反应，这对于新型衣原体DNA疫苗的研究具有重要意义。     

Large-scale testing of women in Copenhagen has not reduced the prevalence of Chlamydia trachomatis infections

Objective  To examine the impact of a stable, large-scale enzyme immunoassay (EIA) Chlamydia trachomatis testing situation in Copenhagen, and to estimate the impact of introducing a genomic-based assay with higher sensitivity and specificity.
Methods  Over a five-year study period, 25 305–28 505 women screened for chlamydial infection each year, corresponding to 19.3% of the female population in Copenhagen, Denmark, were analyzed.
Results  The C. trachomatis age-specific examination percentage and age-specific positive percentage were unchanged during the study period. For EIA, the age-specific positive predictive value of a test decreased from 94% at age 17 to only 50% at age 34 years. Irrespective of the choice of diagnostic test, only about 30% of chlamydial infections would be diagnosed, given current strategies.
Conclusion  Although genomic detection assays will increase the positive and negative predictive values of the Chlamydia test result, new screening strategies for both men and women in younger age groups will be necessary if chlamydial infections are to be curtailed.     

Role of natural killer cells in infection with the mouse pneumonitis agent (murine Chlamydia trachomatis).

Natural killer (NK) activity is increased in both spleen and lung early in pulmonary infection by murine Chlamydia trachomatis in both susceptible nude and resistant heterozygous (nu/+) mice. Ablation of the rise in NK activity by giving the mice antiasialo GM-1 antibody or stimulation of NK activity by immunomodulators did not affect quantitative tissue counts of the mouse pneumonitis biovar of C. trachomatis or significantly affect survival. Studies are needed to further define the role of NK cells in host defense, immunoregulation, and immunopathology during chlamydial infection.     

Analysis of the immune response in mice following intrauterine infection with the Chlamydia trachomatis mouse pneumonitis biovar.

A Swiss Webster white mouse model of salpingitis was used to characterize the immune response following an intrauterine infection with the Chlamydia trachomatis mouse pneumonitis biovar. Western blot (immunoblot) analyses of the serum samples showed that the immunodominant bands corresponded to molecular masses of 72, 60, 42, and 28 kDa and to the lipopolysaccharide. Antibodies to the 60-kDa heat shock protein and to the 60-kDa cysteine-rich protein were detected at 2 and 3 weeks postinfection, respectively. Neutralization was observed in an in vitro assay with serum samples as early as the 3rd day postinfection and remained high for the 7 weeks of observation. The mice were mated in the 7th week following infection. Of the infected experimental mice, 71.4% were found to be either unilaterally or bilaterally infertile, whereas only 27.4% of the noninfected control mice were found to be infertile.     

Circulating antibodies to a conserved epitope of the Chlamydia trachomatis 60-kDa heat shock protein is associated with decreased spontaneous fertility rate in ectopic pregnant women treated by salpingectomy

PROBLEM: This prospective study was aimed to evaluate whether non-invasive clinical and serologic parameters of tubal disease are predictive for subsequent spontaneous conception and pregnancy outcome after first episode of ectopic pregnancy (EP). METHOD OF STUDY: Overall, 144 women aged <35 years were enrolled. Outcome of subsequent spontaneous conception was analyzed after 3 years and compared with clinical parameters and antibody responses to Chlamydia trachomatis and epitopes of the 60-kDa chlamydial heat shock protein (CHSP-60). RESULTS: Antibody response to a conserved epitope of CHSP-60 (amino acids, aa 260-277) was independently correlated with both decreased spontaneous conception and term delivery rates (adjusted odds ratios, OR: 3.6 and 5.4, respectively). CONCLUSION: Presence of circulating antibodies to a conserved epitope of the CHSP-60 is associated with a lower spontaneous conception rate, and increased likelihood of adverse pregnancy outcome in women treated by salpingectomy for first episode of EP.     

Ectopic lymphoid tissue formation in the lungs of mice infected with Chlamydia pneumoniae is associated with epithelial macrophage inflammatory protein-2/CXCL2 expression

Infection with Chlamydia pneumoniae (Cp) accounts for around 10% of community acquired bacterial pneumonia and has been associated with other chronic inflammatory conditions. We describe a C57/Bl6 murine model of Cp lung infection characterized by a dose‐dependent, resolving neutrophilia followed by lymphocytic infiltration of the lungs. By 21 days post‐infection, mice exhibit a T helper type 1 (Th1) polarized serum antibody response with local mucosal antibody secretion and organization of ectopic lymphoid tissue which persisted in the absence of detectable Cp DNA. Macrophage inflammatory protein (MIP)‐2/CXCL2, which recruits neutrophils and lymphocytes and is associated with ectopic lymphoid tissue formation, was secreted in the lungs post‐infection. In vitro, lung epithelial cells up‐regulated MIP‐2/CXCL2 in response to both rough lipopolysaccharide (reLPS) and Cp infection. We conclude that Cp infection can have long‐term inflammatory effects on tissue that persist after clearance of active infection.     

Mice immunized with a chlamydial extract have no increase in early protective immunity despite increased inflammation following genital infection by the mouse pneumonitis agent of Chlamydia trachomatis.

We have determined that immunization with a detergent extract of the mouse pneumonitis agent of Chlamydia trachomatis fails to induce a protective inflammatory immune response following genital infection by C. trachomatis. We demonstrated that mice immunized with the detergent extract have increased cutaneous delayed-type hypersensitivity and increased splenic T-cell proliferation in response to the chlamydial extract. After genital infection by C. trachomatis, extract-sensitized mice had significantly increased genital inflammation (P = 0.044) compared with controls. The inflammation was characterized by significantly increased eosinophils in the genitalia (P < 0.0005) and increased genital edema (P < 0.0005). However, the increased genital inflammation of extract-sensitized mice provided no increase in protection against infection (P = 0.92).     

Mycobacteria lacking the RD1 region do not induce necrosis in the lungs of mice lacking interferon-gamma

The genetic region of difference 1 (RD1) in Mycobacterium tuberculosis has recently been hypothesized to encode for proteins that are cytotoxic to the host cell in nature. We demonstrate here that while M. tuberculosis grew progressively in the lungs of gene disrupted mice (GKO) unable to produce interferon-gamma (IFN-gamma), similar mice infected instead with M. bovis bacillus Calmette-Guérin (BCG) reproducibly exhibited an obvious slowing of the disease after about 20 days. Closer examination of BCG-infected GKO mice showed a florid granulomatous inflammation in the lungs, whereas similar mice infected with M. tuberculosis exhibited wholesale progressive necrosis. In the BCG-infected GKO mice large numbers of activated effector T cells, some strongly positive for the cytokine tumour necrosis factor, as well as activated natural killer cells accumulated in the lungs. To further test the hypothesis that the differences observed were directly associated with the loss of the RD1 region, it was then shown that a mutant of M. tuberculosis lacking RD1 grew progressively in both normal and GKO mice but failed to induce any degree of necrosis in either animal despite reaching similar levels in the lungs. However, when mice were infected with this mutant, in which the RD1 region had been restored by complementation, wholesale necrosis of the lungs again occurred. These data support the hypothesis that proteins encoded in the RD1 region are a major cause of necrosis and contribute significantly to the pathogenesis of the disease.     

Immunology: Autoimmunity to spermatozoa, asymptomatic Chlamydia trachomatis genital tract infection and {gamma}{delta} T lymphocytes in seminal fluid from the male partners of couples with unexplained infertility

The relationship between an undetected, asymptomatic Chlamydiatrachomatis genital tract infection, the concentration of andb T cells in semen and sperm autoimmunity was examined in 48male partners of couples with unexplained infertility. ImmunoglobulinA (IgA) antibodies to C.trachomatis were detected in seminalfluids from 14 (29.2%) of the men. Only four of these were positivefor circulating anti-chlamydial IgA, suggesting that the stimulusfor antibody production was within the genital tract. In contrast,four men were positive for anti-chlamydial IgG in their semen;all were also seropositive for anti-chlamydial IgG. T lymphocytesbearing the and antigen receptors were present in every semensample. Men with seminal anti-chlamydial IgA, however, had significantly(P = 0.035) elevated semen T cell concentrations (median 3100cells/ml) than did men lacking this antibody (median 1400 cells/ml);concentrations of T cells were comparable in both groups. Genitaltract sperm autoimmunity, as shown by antibodies bound to motileejaculated spermatozoa, was detected in 13 (27.1%) men. Thepresence of these antibodies was associated with elevated concentrationsof both (median 4200 versus 700 cells/ml) and (median 5000versus 850 cells/ml) T cells (P = 0.0002 and 0.0001 respectively).Men with antisperm antibodies only in their serum had seminalT cell concentrations comparable with men testing negative forantisperm antibodies. Anti-chlamydial IgA was identified insemen from four of 10 men with IgA bound to their spermatozoaand in none of the men with only spermatozoabound IgG. Therewas no relationship between sperm quality and the occurrenceof seminal IgA antibodies to either C.trachomatis or spermatozoa.An asymptomatic C.trachomatis infection activates T cells withinthe male genital tract, which may lead to antisperm antibodyformation and immune-mediated infertility.     

Insight into the role of TSLP in inflammatory bowel diseases

Proinflammatory cytokines are thought to modulate pathogeneses of various inflammatory bowel diseases (IBDs). Thymic stromal lymphopoietin (TSLP), which has been studied in various allergic diseases such as asthma, atopic dermatitis (AD) and eosinophilic esophagitis (EoE), has been less considered to be involved in IBDs. However, mucosal dendritic cells (DCs) induced by various cytokines including TSLP were reported to cause polarization of T cell toward Th2 response, the differentiation of regulatory T-cell (Treg), and secretion of IgA by B cells. In this review, we discuss the concept that decreased TSLP has the potential to accelerate the development of Th1 response dominant diseases such as the Crohn's disease (CD) while increased TSLP has the potential to lead to a development of Th2 cell dominant diseases such the ulcerative colitis (UC). To examine TSLP's role as a potential determining factor for differentiating UC and CD, we analyzed the effects of other genes regulated by TSLP in regards to the UC and CD pathogeneses using data from online open access resources such as NetPath, GeneMania, and the String database. Our findings indicate that TSLP is a key mediator in the pathogenesis of IBDs and that further studies are needed to evaluate its role.