Serological responses of patients with ectopic pregnancy to epitopes of the Chlamydia trachomatis 60 kDa heat shock protein

Clinical and histopathological correlations of immunoreactivity to Chlamydia trachomatis and to epitopes of the C. trachomatis 60 kDa heat shock protein (hsp60) among women with ectopic pregnancy were evaluated in a case-control study. Serological responses to 13 synthetic peptides corresponding to major epitopes of the chlamydial hsp60 were determined in 67 women treated for ectopic pregnancy and 45 women with uncomplicated pregnancy in utero. Plasma cell salpingitis was detected in 29 (43.3%) of the ectopic patients. Its presence correlated with antibodies to two hsp60 epitopes, encompassing amino acids 260-271 and 411-422 (P = 0.02). Antibodies to these two epitopes, along with five other epitopes, also correlated with peritubal adhesion formation in ectopic pregnant patients (P < 0.01). Antibodies to epitopes 260-271 and 188-199 also correlated with a history of pelvic inflammatory disease (PID; P = 0.05). Patients with ectopic pregnancy were also more likely than their intrauterine pregnant controls to have present anti-chlamydial immunoglobulin G (P < 0.005). Women positive for both C. trachomatis and hsp60 epitope antibodies had an increased prevalence over controls of salpingitis, pelvic adhesions or history of PID (P < 0.05). In contrast, patients who were positive for only C. trachomatis antibodies or only hsp60 epitope antibodies did not differ from antibody-negative patients in each of these categories.     

Gene knockout B cell-deficient mice demonstrate that B cells play an important role in the initiation of T cell responses to Chlamydia trachomatis (mouse pneumonitis) lung infection

T cell-mediated immunity as measured by delayed-type hypersensitivity, and IFN-gamma production has been shown to be critical for host defense against Chlamydia trachomatis infection in both human and animal studies. Using gene-targeted B cell-deficient mice, we examined the role of B cells in protective immunity to C. trachomatis (mouse pneumonitis) (MoPn) lung infection. B cell-deficient mice were observed to have a significantly higher mortality rate and in vivo chlamydial growth than did wild-type mice following MoPn lung infection. Interestingly, B cell-deficient mice not only lacked Ab responses but also failed to mount an efficient delayed-type hypersensitivity response following chlamydial lung infection. In contrast to results obtained from MoPn-infected wild-type C57BL/6 mice, spleen cells from infected B cell-deficient mice failed to produce Th1-related (IFN-gamma) or Th2-related (IL-6 and IL-10) cytokines after Chlamydia-specific in vitro restimulation. Moreover, unlike wild-type mice, B cell-deficient mice were not immune to rechallenge infection following recovery from primary chlamydial infection. The data indicate that B cells play an important role in host defense to primary and secondary chlamydial infection and suggest that B cells are crucial for the initiation of early T cell responses to chlamydial infection. This study provides evidence for the role of B cells in the in vivo priming of T cells during infection with the intracellular bacterial pathogen, C. trachomatis.     

Cost analysis for total hip arthroplasty by measurement of time and material expenditure

BACKGROUND AND OBJECTIVES: An estimated 4 million new cases of chlamydial infection occur each year. This experiment assessed the effects of a vaginally applied gel formulation of 0.25% chlorhexidine gluconate on chlamydial infection and on the vaginal ecosystem. STUDY DESIGN: Twelve monkeys were treated with a single application of 0.25% chlorhexidine gluconate. These animals were assessed for changes in vaginal flora before and at 30 minutes, 1 day, and 2 days postapplication by microbiologic analysis. Cervical and vaginal tissues were assessed by colposcopy at each time point. Five monkeys received a single application of 0.25% chlorhexidine gluconate gel followed (30 minutes) by a cervical inoculation with Chlamydia trachomatis. Four monkeys were inoculated with Chlamydia only. Cervicovaginal tissues were assessed via modified colposcopy, vaginal swabs were collected for assessment of vaginal flora, and cervical swabs were collected for detection of Chlamydia (culture/ligase chain reaction) at baseline and days 1, 2, and 7 postinoculation. RESULTS: Changes in vaginal flora were minimal in all monkeys. Application of 0.25% chlorhexidine gluconate did not affect adversely vaginal colonization by lactobacilli. All chlamydial infection control monkeys were infected, whereas none of the five monkeys pretreated with chlorhexidine gluconate were positive for C. trachomatis by culture or ligase chain reaction. Colposcopic observations remained largely unchanged in all groups. CONCLUSIONS: A 0.25% chlorhexidine gluconate gel was protective against chlamydial infection in all animals tested, had no adverse effect on the vaginal flora, and had minimal effect on cervicovaginal tissues after a single application.     

Response of Chlamydia trachomatis serovar E to iron restriction in vitro and evidence for iron-regulated chlamydial proteins

Iron is a well-established mediator of virulence in several bacterial pathogens, yet little is known about the role of iron in infectious disease processes caused by obligate intracellular bacterial pathogens. In this study, the effect of iron limitation was examined for the sexually transmitted infectious agent Chlamydia trachomatis in an in vitro model of human genital infection using the intracellular iron-chelating reagent deferoxamine mesylate (Desferal). Iron restriction caused a significant reduction in infectivity of C. trachomatis elementary bodies (EB) harvested from Desferal-exposed polarized epithelial cells when compared to that of EB harvested from iron-sufficient control cell cultures. Replacement of the Desferal exposure medium with medium containing iron-saturated transferrin restored chlamydial infectivity, whereas replacement with growth medium alone had no effect. The following three prominent morphological features were observed by electron microscopic examination of chlamydia-infected cells exposed to Desferal: (i) inclusions containing chlamydiae greatly delayed in maturation, (ii) substantial blebbing within chlamydial inclusions, and (iii) electron-dense material surrounding inclusions. Protein analyses of highly purified EB by two-dimensional polyacrylamide gel electrophoresis revealed that there were at least 19 candidate iron-repressible proteins in C. trachomatis and at least one protein which was iron inducible. One putative iron-repressible protein was confirmed by Western blot (immunoblot) analysis to be the chlamydial heat shock protein 60 (hsp60). The enhanced production of this antigen by chlamydiae as a result of iron limitation is of particular importance since there is a well-documented association between chlamydial hsp60 and destructive immunopathological sequelae in infected patients.     

Cell-mediated immune response to the recombinant 57-kDa heat-shock protein of Chlamydia trachomatis in women with salpingitis

Peripheral blood mononuclear cells (PBMC) from 9 of 18 women with laparoscopy-verified salpingitis proliferated in response to recombinant Chlamydia trachomatis 57-kDa heat shock protein (hsp). In contrast, PBMC from 0 of 10 women with cervicitis, 1 of 5 women with recurrent abortions, and 3 (7.1%) of 42 healthy reproductive-age women were responsive to hsp (P < .001). After passage of the hsp through an endotoxin-removing column, PBMC from 6 of 14 additional women with salpingitis were responsive to hsp, while those from 10 controls, including the 3 previously positive women, were negative. PBMC from all patients responsive to the chlamydial hsp were unresponsive to Mycobacterium bovis 65-kDa hsp. PBMC from 6 of the 15 women with a positive hsp-induced lymphocyte response were unresponsive to C. trachomatis elementary bodies. Induction of a cell-mediated immune response to the chlamydial 57-kDa hsp is a common feature of an upper genital tract infection but does not appear to be limited to women with apparent chlamydial infections.     

Chlamydial genital infections and laparoscopic findings in infertile women

Several studies have shown that previous chlamydial genital infection, reflected by serological markers, is strongly associated with tubal damage leading to tubal infertility. In 105 women undergoing laparoscopy, multiple samples were collected from the lower (urethra and cervix) and upper (endometrium, peritoneal fluid, tubal lumen) genital tract, in order to isolate Chlamydia trachomatis in cell culture. Chlamydia trachomatis was isolated from at least one site in 13 (30.9%) of 42 infertile women with tubal infertility, in 5 (12.1%) of 41 women with unexplained infertility, in 1 of 4 women affected by acute salpingitis and in 1 (5.5%) of 18 women with endometriosis or uterine malformations. The latter group was the control group. Thirteen (65%) of the 20 positive women harboured Chlamydia trachomatis in their upper genital tract alone and 16 women were positive in one or both tubes. Only one of the positive women showed laparoscopic signs of acute pelvic infection. Four of the 5 positive women with unexplained infertility harboured Chlamydia trachomatis in the tubal lumen. This study confirms that chlamydial infection is strongly associated with tubal damage. It suggests that cervical cultures are inadequate for excluding a tubal infection and that chlamydial colonization of the tubal mucosa is possible in the absence of symptoms and laparoscopic signs of active infection.     

Anthropometric, strength, endurance and flexibility characteristics of elite and recreational climbers

Chlamydia trachomatis is a primary cause of acute or silent salpingitis leading to infertility and ectopic pregnancy. The C. trachomatis epidemic, undiscovered in most cases, spreads, mostly in adolescents, during the years following the onset of sexual activity. As opposed to gonococcal infection which has greatly decreased, C. trachomatis cervical and urethral infection is common in young occidentals. More then 30 different studies covering 200-12,000 subjects screened in family planning centers, college women and men, students and military recruits in different parts of the USA, in Scandinavian countries and France, indicate a prevalence of 5-20% (mean 10%) in apparently healthy young females < 25 years and 5-10% in males. Female prevalence is strongly related to age, being highest (15-20%) in women < 20 years old. Several cost-benefit analyses show that the total cost of the general screening in young populations, which can easily be carried out for women in family planning centers, could save twice the cost of treatment for pelvic inflammatory disease caused by C. trachomatis and six times the total cost of C. trachomatis epidemics if late sequelae are taken into account (tubal infertility treatment, ectopic pregnancy). There is a debate among authors concerning the relative merits of total screening versus selective screening in family planning centers, the most common opinion being to do a total screening of women < 20 years old and selective screening of women 20-30 years of age with at least one risk factor, the most common risk factors being more than one partner in a year, purulent, cervical discharge, failure to use condoms and use of a contraceptive pill. Although the data clearly show that C. trachomatis screening is cost-effective, conducting of the diagnostic laboratory tests used in such screening programs should be carefully evaluated relative to cost, feasibility, specificity and sensitivity and should be adapted to the presumed prevalence in screened populations.     

Immunological findings in a case of Cogan's syndrome

The immune system of a case of Cogan's syndrome was investigated. A transient depression of the cell-mediated immunity was found. Thus, during the acute stage of the disease, there was a depression of the cutaneous delayed hypersensitivity reaction to PPD and a decrease in the number of T cells. There were also signs of complement consumption. A possible pathogenesis based on immune complexes due to a preceding viral infection is discussed.     

Expression of recombinant DNA introduced into Chlamydia trachomatis by electroporation

Electroporation was used to introduce DNA into the elementary bodies of the obligate parasitic bacterium Chlamydia trachomatis. The source of DNA for these experiments was the chimeric plasmid pPBW100, which was constructed from the well-characterized 7.5-kb plasmid of C. trachomatis and the Escherichia coli plasmid pBGS9. To select directly for C. trachomatis carrying pPBW100, an in-frame gene fusion between the chlamydial promoter P7248 and a promoterless chloramphenicol acetyltransferase (cat) cassette was incorporated into the plasmid. After infection of McCoy cells with electroporated elementary bodies containing pPBW100, the following were observed: (i) the plasmid DNA was detected inside the chloramphenicol-resistant chlamydial inclusions by in situ and Southern hybridization analyses; (ii) both physical and biochemical evidence showed that chloramphenicol acetyltransferase was synthesized by the electroporated C. trachomatis; (iii) expression of P7248::cat was developmentally regulated and occurred during the early stages of chlamydial reticulate body development; and (iv) although the expression from P7248::cat was mainly transient, there were rare instances where chloramphenicol-resistant C. trachomatis were observed after four passages.     

Local immune responses to Chlamydia pneumoniae in the lungs of BALB/c mice during primary infection and reinfection

Cell-mediated immune (CMI) responses play a major role in protection as well as pathogenesis of many intracellular bacterial infections. In this study, we evaluated the infection kinetics and assessed histologically the lymphoid reactions and local, in vitro-restimulated CMI responses in lungs of BALB/c mice, during both primary infection and reinfection with Chlamydia pneumoniae. The primary challenge resulted in a self-restricted infection with elimination of culturable bacteria by day 27 after challenge. A mild lymphoid reaction characterized the pathology in the lungs. In vitro CMI responses consisted of a weak proliferative response and no secretion of gamma interferon (IFN-gamma). The number of lung-derived mononuclear cells increased substantially during the primary infection; the largest relative increase was observed in B cells (B220(+)). After reinfection, the number of lung-derived mononuclear cells increased further, and the response consisted mainly of T cells. The reinfection was characterized in vivo by significant protection from infection (fewer cultivable bacteria in the lungs for a shorter period of time) but increased local lymphoid reaction at the infection site. In vitro, as opposed to the response in naive mice, acquired immunity was characterized by a strongly Th1-biased (IFN-gamma) CMI response. These results suggest that repeated infections with C. pneumoniae may induce Th1-type responses with similar associated tissue reactions, as shown in C. trachomatis infection models.     

Antibody to chlamydial hsp60 predicts an increased risk for chlamydial pelvic inflammatory disease

To determine whether serum antibody to Chlamydia trachomatis antigens alters the risk of C. trachomatis pelvic inflammatory disease (PID), 280 female sex workers were prospectively evaluated over a 33-month period for incident C. trachomatis and Neisseria gonorrhoeae cervical infection and for clinical PID. At enrollment, women were tested for antibody to C. trachomatis elementary bodies by an indirect microimmunofluorescence assay and to recombinant chlamydial hsp60 (Chsp60) by an ELISA format. At each follow-up visit, women were tested for cervical chlamydial and gonococcal infection and were identified as having clinical PID if they complained of lower abdominal pain and were found to have uterine and adnexal tenderness on pelvic examination. The data demonstrate that antibody to Chsp60 predicts a 2- to 3-fold increased risk for C. trachomatis PID.     

Postoperative ileus as an indication for a 2d operation during the early postoperative period

The phagocytic and bactericidal activities to Salmonella pullorum (strain 9-25) or Salmonella senftenberg (strain 99D) were examined in chicken splenic phagocytes from 0-day-old to 2-month-old chickens. The phagocytic activity against S pullorum increased in splenic phagocytes from chickens older than 7 days, but significant changes in activity against S senftenberg were not observed during the experimental period. The bactericidal activity of splenic phagocytes against S senftenberg was higher than that of phagocytes against S pullorum during the same period. Increase of the bactericidal activity against S pullorum was observed with increasing age, but the activity of the splenic phagocytes from 0-day-old chickens against S senftenberg was similar to that of the phagocytes from 2-month-old chickens. Although delayed hypersensitivity was confirmed by delayed wattle reaction in 2-month-old chickens sensitized with living S pullorum, the sensitization did not markedly affect phagocytic and bactericidal activities.     

Delayed-type skin reactions in bursectomized or thymectomized chickens

Chickens can easily be induced to develop delayed-type skin reactions to oxazolone when animals are sensitized 7 days before the challenge. The reaction is quantitated by assessing the increase in wattle thickness: maximum reactions occur 24 h after challenge. The reaction is inhibited by neonatal thymectomy or bursectomy; these findings therefore suggest also an important B-derived component in delayed hypersensitivity to oxazolone.     

Studies on local immune response in mouse model of experimental Escherichia coli intrauterine infections

Studies were conducted to elucidate the immune cell response at infection sites by performing immunostaining of immune cells with a monoclonal antibody in an experimental Escherichia coli (E. coli) mouse uterine infection model. 1. The incidence of uterine infection by E. coli decreased with the passage of time: 4/4 on Day 1, 4/6 on Day 3, 2/6 on Day 7, and 1/6 on each of Days 14 and 21. It was surmised that clearance of the bacteria from the infection sites was being carried out by immune cells. 2. Beginning from infection Day 1, the infected uterine tissue was observed to undergo a moderate degree of invasion by neutrophils, macrophages, CD4+ T cells, CD8+ T cells and IgA+ B cells. Then, beginning from infection Day 3, there was a mild degree of invasion of the infected uterine tissue by IgM+ B cells and IgG+ B cells. The number of neutrophils in the tissue decreased beginning from infection Day 14, but the degree of invasion of the infected tissue by the other kinds of immune cells remained almost constant through infection Day 21. 3. A comparison was made of the immune responses to local infection by E. coli, and Chlamydia trachomatis (C. trachomatis), an intracellular parasite. It was found that the invasion of the infection site by immune cells occurred earlier in the case of E. coli infection than C. trachomatis infection. In addition, the C. trachomatis infection site was observed to contain greater numbers of macrophages and CD8+ T cells play important roles in the immune defense at sites of infection by C. trachomatis.     

Detection of Chlamydia trachomatis in urine samples by nucleic acid tests: comparison with culture and enzyme immunoassay of genital swab specimens

Two commercially available nucleic acid-based tests, ligase chain reaction (LCR; Abbott Laboratories) and PCR (Roche Diagnostics), for the detection of Chlamydia trachomatis in male and female urine samples were compared with culture and enzyme immunoassay (EIA) (Microtrak; Syva) for C. trachomatis detection in genital samples. The samples were collected from 1,005 patients who attended a sexually transmitted disease clinic. In this study population, the prevalence of the infection was 4%. Specimens which were reactive in any of the tests were retested with a different PCR test using primers directed against the major outer membrane protein gene. With a "gold standard" of a positive culture, or any other positive test result if it was confirmed by an independent test, the Roche PCR (95% sensitive, 99.9% specific) was more sensitive than the LCR (75% sensitive, 100% specific) (chi2, P < 0.0001) while both tests were more sensitive than culture (58% sensitive, 100% specific) or EIA (45% sensitive, 100% specific) (chi2, P < 0.001). The Roche PCR and Abbott LCR tests of urine identified 65% and 30% more positive patients, respectively, than did testing by culture of urethral or cervical specimens. Nucleic acid testing of urine specimens for C. trachomatis is a more sensitive and convenient method for the detection of genital infection.     

Mouse NK1.1+ T cells: a new family of T cells

A ligase chain reaction (LCR) DNA amplification assay that targeted the cryptic plasmid of Chlamydia trachomatis was developed to detect C. trachomatis urogenital tract infection. The objectives of this study were to determine the cutoff and analytic performance of the LCR assay and to characterize the effectiveness of its postdetection contamination control method. The assay's cutoff was determined after receiver-operator characteristic (ROC) analysis of 4660 clinical data points. The assay detected one infectious unit per reaction of each of the 15 C. trachomatis serovars and did not cross-react with 13 Chlamydia pneumoniae strains, 13 Chlamydia psittaci strains, and 87 other bacteria, fungi, parasites, or viruses. In addition, the assay did not detect 77 processed urine specimens collected from patients with urinary tract infections caused by yeast or bacteria other than C. trachomatis. The assay was sufficiently precise to detect consistently two infectious units of C. trachomatis per reaction. False-positive assay results attributable to contamination with amplified product were minimized by the use of standard procedures as well as by a postdetection chemical inactivation method that could reduce the amount of amplified LCR product by a factor of > or = 10(7).     

Effects of SL-probiotic preparation on the body weight and phagocytosis of white mice

Specimens from 15 young patients presenting with acute epididymitis were tested for the presence of Chlamydia trachomatis by an enzyme immunoassay (EIA), polymerase chain reaction (PCR), and for other bacteria by standard laboratory techniques. C. trachomatis urethral infection was detected in 3 patients by an EIA test of the urethral swabs (20%) and in 13 patients by the PCR (87%). This difference in detection rate was statistically significant (p < 0.005). Thirteen specimens were positive by the PCR, but only three of them were positive by the EIA method. These findings indicate that the PCR assay is a highly sensitive assay for the detection of C. trachomatis in male urine specimens and provides a noninvasive technique for routine screening of chlamydia infection in the patient with acute epididymitis.     

Total knee arthroplasty without patellar resurfacing in active and overweight patients

Chlamydia trachomatis is a frequent sexually transmitted disease. The diagnosis of C. trachomatis infection by cytology is controversial. We compared the ability of Papanicolaou (Pap) smears to detect C. trachomatis infection with antigen detection (enzyme immunoassay; EIA) and polymerase chain reaction (PCR). One hundred sixty-seven women attending a therapeutic abortion clinic were enrolled in the study. Endocervical samples were first collected for EIA and PCR, and then Pap smears were prepared for cytologic evaluation. Eight patients were excluded from the study due to the lack of an endocervical component. The criteria established by Gupta and associates (Diagn Cytopathol 1988;4:224-229; Acta Cytol 1979;23:315-320) were used in this study to assess the specificity and sensitivity of the Pap smear in recognizing C. trachomatis infection. After EIA testing, the remaining sample was subjected to phenol-chloroform extraction to purify the DNA and then tested by PCR. Positive PCR samples were subjected to repeat phenol-chloroform and retested to confirm the positive result. Using a confirmed PCR or a blocked EIA as the extended gold standard, the incidence of C. trachomatis infection was 9.4%. Fifteen of the 159 cases reviewed were positive by extended gold standard. Thirteen (86.7%) of those 15 cases were interpreted as negative by cytology (false-negatives), and two (13.3%) cases were positive. Of the remaining 144 cases, 14 cases (9.7%) were interpreted as positive by cytology (false-positives) but were not confirmed by the extended gold standard. Ten (66.7%) of the 15 cases confirmed by the extended gold standard were interpreted as negative by EIA (false-negatives), and five (33.3%) were positive. There were no false-positives by EIA. In this study, the sensitivity and the specificity for cytology were 13.3% and 90.3%, respectively. The positive predictive value was 12.5%, and the negative predictive value for cytology was 90.9%. The sensitivity and the specificity for EIA were 33.3% and 100%, respectively. The positive predictive value was 100%, and the negative predictive value for EIA was 93.5%. Both EIA and cytology are insensitive methods compared with PCR. Based on these data, cytology should not be used to diagnose C. trachomatis infection in an asymptomatic female population with a moderate risk of C. trachomatis infection.     

Alteration of activities of hepatic antioxidant defence enzymes in developing chick embryos after glucocorticoid administration--a factor to produce some adverse effects?

Nickel is a strong biological sensitizer and consequently may induce a delayed hypersensitivity reaction (type IV immune response). Because nickel is a component of the majority of the orthodontic alloys, the objectives of this cross-sectional study were to determine the prevalence of nickel hypersensitivity reaction before, during, and after orthodontic therapy with conventional stainless steel brackets and wires; to evidence the induction of this reaction by the orthodontic appliances; and to characterize the nickel hypersensitive persons. Nickel patch tests and a questionnaire were used to evaluate the hypersensitivity to this metal. The total sample consisted of 170 patients, 105 females and 65 males, from the orthodontic department at Bauru Dental School, University of S?o Paulo. They were divided into three groups as follows: A (n = 60), patients before the beginning of orthodontic therapy; B (n = 66), patients currently undergoing orthodontic treatment, and C (n = 44), patients who had undergone orthodontic treatment previously. The chi-square test (chi2) showed an allergic reaction in 28.3% of the total sample with 23% female and 5.3% male. This indicated a gender difference (chi2 = 10.75, p < 0.001). There was a positive association between nickel hypersensitivity and previous personal allergic history to metals (chi2 = 34.88, p < 0.0001) as well as with the daily use of metal objects (chi2 = 11.95, p < 0.0005). There was no statistically significant difference in the prevalence of contact dermatitis among the three groups (chi2 = 0.39, p = 0.848). This suggests that orthodontic therapy with conventional stainless steel appliances does not initiate or aggravate a nickel hypersensitivity reaction.     

A prospective study of transvaginal hydrosonography in the evaluation of abnormal uterine bleeding

OBJECTIVE: To perform a cost-benefit analysis of a Chlamydia trachomatis screening program based on first-void urine testing of asymptomatic women using a polymerase chain reaction (PCR) test. METHODS: A decision tree was developed. Selected variables based on assumptions were subjected to sensitivity analyses to make the model accurate and defensible. RESULTS: Screening for chlamydial infections using the PCR test was shown to be cost-effective even in low-prevalence populations. Compared with a symptom-driven no-screening situation, a universal C trachomatis screening program using the PCR test would save money, in terms of direct cost, when the baseline prevalence of C trachomatis infection exceeds 3.9%. CONCLUSION: Cost analyses are still rare among trials that compare pharmacologic or procedural health care interventions. Socioeconomic studies linking secondary prevention of C trachomatis infection and infertility and adverse pregnancy outcome are needed to convince public health authorities of the need for and the benefit of such programs.