Heparan sulfate-like glycosaminoglycan is a cellular receptor for Chlamydia pneumoniae.

Chlamydia pneumoniae is an important human intracellular pathogen; however, the pathogenesis of C. pneumoniae infection is poorly understood, and the bacterial adherence mechanism to host cells is unknown. This study examined the role of glycosaminoglycans (GAGs) in the adhesion of C. pneumoniae to eukaryotic cells. Heparin and heparan sulfate were found to inhibit the attachment of C. pneumoniae to human epithelial cells. Reduction in infectivity resulted from the binding of heparin to the organism. Enzymatic removal of heparan sulfate moieties from the host cell surface led to a marked decrease in C. pneumoniae infectivity. Mutant CHO cell lines that were defective in heparan sulfate biosynthesis were less susceptible to C. pneumoniae infection than was the wild-type cell line. However, preincubation of the GAG-deficient CHO cells with exogenous heparin greatly increased infectivity.

Infection and activation of airway epithelial cells by Chlamydia pneumoniae.

The activation of primary human airway epithelial cells (HAECs) and of the bronchial epithelial cell line BEAS-2B by Chlamydia pneumoniae, an important respiratory pathogen, was characterized. A time-dependent enhanced release of interleukin (IL)-8 and prostaglandin-E(2) and an increased expression of the epithelial adhesion molecule intercellular adhesion molecule-1 (ICAM-1), followed by subsequent transepithelial migration of polymorphonuclear neutrophils (PMN), were also demonstrated. The transepithelial PMN migration could be blocked by an anti-ICAM-1 monoclonal antibody (MAb) but not by MAbs against IL-8. In addition, there was an enhanced C. pneumoniae-mediated activation of NF-kappaB within 30-60 min in HAECs and BEAS-2B, which was followed by increases in mRNA synthesis of IL-8, ICAM-1, and cyclooxygenase-2, with maximal effects occurring 2 h after infection. Thus, C. pneumoniae infects and activates HAECs and BEAS-2B and therefore may be able to trigger a cascade of pro- and anti-inflammatory reactions during chlamydial infections.

A search for beta-lactamase in chlamydiae, mycoplasmas, planctomycetes, and cyanelles: bacteria and bacterial descendants at different phylogenetic positions and stages of cell wall development.

Bacteria from different phylogenetic positions such as chlamydiae, mycoplasmas, planctomycetes and also endosymbiotic murein-containing cyanelles were investigated for the production of beta-lactamases. No beta-lactamase activity was found in bacteria lacking murein such as Chlamydia pneumoniae, Mycoplasma pneumoniae, Pirellula marina and Planctomyces maris. In the murein-containing cyanelles of Cyanophora paradoxa no beta-lactamase activity could be detected.

Chlamydia pneumoniae infection is not an independent risk factor for arterial disease.

Appreciation of atherosclerosis as an infectious disease has fostered interest in the role that Chlamydia pneumoniae may play in atheroma development. Although data from seroepidemiological and experimental studies have established an association between the pathogen and atherosclerosis, little is known about how the organism contributes to lesion development. Atherosclerosis is a complex disease process and the role of any pathogen must be considered in the context of other risk factors. Here we focus on the relationship between Chlamydia pneumoniae and conventional risk factors for atherosclerosis. There is evidence for a strong association between chronic infection with Chlamydia pneumoniae and smoking as well as high serum cholesterol. It is concluded from the present data that chronic infection with the pathogen is not an independent risk factor for atherosclerosis.

Antibody response to the 60-kDa heat-shock protein of Chlamydia pneumoniae in patients with coronary artery disease.

Serum specimens from 752 individuals undergoing coronary arteriography were examined for antibodies to Chlamydia pneumoniae. Patients with coronary artery disease (CAD) were more likely to have IgG antibodies to C. pneumoniae than were individuals without CAD (60% vs. 52%; P=.007; odds ratio, 1.8; 95% confidence interval, 1. 17-2.77). Antibodies to recombinant hsp60 of C. pneumoniae were found with nearly the same frequency in patients with CAD and individuals without CAD (29% vs. 30%; P=.751). There was no association between chlamydial hsp60 antibodies and the severity of CAD or a previous myocardial infarction. Patient sera reacted most frequently to C. pneumoniae proteins of 17, 38, 40, 58, and 60/62 kDa. Reactivity to these proteins was not different between patients with and without CAD. Study results indicate that neither antibodies to chlamydial hsp60 nor antibodies to other C. pneumoniae proteins are useful for discriminating between seropositive patients with and without CAD.

Low prevalence of Chlamydia pneumoniae in atherectomy specimens from patients with coronary heart disease.

Coronary atherectomy specimens from 50 patients with coronary heart disease were examined for the presence of Chlamydia pneumoniae by two different methods of polymerase chain reaction (PCR) and by in situ hybridization. C. pneumoniae DNA was detected by PCR in atherosclerotic plaques of four patients (8%). Two patients' coronary atheromas were positive, both by a single-step 16S rRNA-based PCR and by an omp1-based nested PCR. The other two patients' specimens were positive only by the nested PCR. In contrast, C. pneumoniae was not detected by in situ hybridization in any of the cardiovascular tissues tested. Of three patients with evidence of C. pneumoniae in coronary atheromas, two had an antibody titer of 1:32 and the third had no specific antibodies detectable. Results of this study demonstrate a low prevalence of C. pneumoniae DNA in coronary atheromas. These findings do not support the hypothesis that the organism plays a major role in atherogenesis.

Signal transduction pathways activated in endothelial cells following infection with Chlamydia pneumoniae.

Chlamydia pneumoniae is an important respiratory pathogen. Recently, its presence has been demonstrated in atherosclerotic lesions. In this study, we characterized C. pneumoniae-mediated activation of endothelial cells and demonstrated an enhanced expression of endothelial adhesion molecules followed by subsequent rolling, adhesion, and transmigration of leukocytes (monocytes, granulocytes). These effects were blocked by mAbs against endothelial and/or leukocyte adhesion molecules (beta1 and beta2 integrins). Additionally, activation of different signal transduction pathways in C. pneumoniae-infected endothelial cells was shown: protein tyrosine phosphorylation, up-regulation of phosphorylated p42/p44 mitogen-activated protein kinase, and NF-kappaB activation/translocation occurred within 10-15 min. Increased mRNA and surface expression of E-selectin, ICAM-1, and VCAM-1 were noted within hours. Thus, C. pneumoniae triggers a cascade of events that could lead to endothelial activation, inflammation, and thrombosis, which in turn may result in or may promote atherosclerosis.

Rapid detection of Chlamydia pneumoniae by PCR-enzyme immunoassay.

Chlamydia pneumoniae is an important human respiratory pathogen. Laboratory diagnosis of infection with this organism is difficult. To facilitate the detection of C. pneumoniae by PCR, an enzyme immunoassay (EIA) for analysis of PCR products was developed. Biotin-labeled PCR products generated from the 16S rRNA gene of C. pneumoniae were hybridized to a digoxigenin-labeled probe and then immobilized to streptavidin-coated microtiter plates. Bound PCR product-probe hybrids were detected with antidigoxigenin peroxidase conjugate and a colorimetric substrate. This EIA was as sensitive as Southern blot hybridization for the detection of PCR products and 100 times more sensitive than visualization of PCR products on agarose gels. The diagnostic value of the PCR-EIA in comparison to cell culture was assessed in throat swab specimens from children with respiratory tract infections. C. pneumoniae was isolated from only 1 of 368 specimens tested. In contrast, 15 patient specimens were repeatedly positive for C. pneumoniae by PCR and Southern analysis. All of these 15 specimens were also identified by PCR-EIA. Of the 15 specimens positive by 16S rRNA-based PCR, 13 specimens could be confirmed by omp1-based PCR or direct fluorescent-antibody assay. Results of this study demonstrate that PCR is more sensitive than cell culture for the detection of C. pneumoniae. The EIA described here is a rapid, sensitive, and simple method for detection of amplified C. pneumoniae DNA.

Experimental genital tract infection with Chlamydia psittaci (GPIC agent) in male rats.

The course of experimental chlamydial infection of the male genital tract was studied. Inoculation of the Chlamydia psittaci agent of guinea pig inclusion conjunctivitis (GPIC agent) into the vas deferens of rats resulted in chlamydial infection of the epididymis, testis and the prostate gland. The inflammatory response was most prominent at 14 days after infection. Chlamydiae were recovered from the epididymides and the prostate glands for up to 90 and 60 days post inoculation, respectively. Histopathological changes associated with chlamydial infection of the epididymis or prostate gland were characterized by intratubular and interstitial purulent inflammation. Chlamydia-specific IgM- and IgG-antibodies were found in sera of nearly all infected animals. Results of this study indicate that this animal model may be useful to study the pathogenesis, immune responses and sequelae of chlamydial infections of the male genital tract.

Tissue penetration of sparfloxacin in a rat model of experimental Escherichia coli epididymitis.

The epididymal, testicular, and prostatic tissue penetration of sparfloxacin, a new quinolone, was assessed in a rat model of acute epididymitis. Seventy-two hours after injection of 0.1 ml (10(6) cfu/ml) of an Escherichia coli suspension into the right epididymis via the right ductus deferens, a single oral dose of sparfloxacin 50 mg/kg body weight was administered. One, 2, 4, 8, 12, and 24 h after administration the animals were sacrificed and the sparfloxacin concentrations and "areas under the curve" (AUC0-24) in both epididymides, both testes, the prostate gland and in the serum were measured by bioassay. The highest mean AUC0-24 was found in the prostate gland, followed by left epididymis, right epididymis, serum, right testis, and left testis (190, 79, 60, 28, 12, and 9 mg/kg x h, respectively). Though there was no statistically significant difference in the sparfloxacin concentration of both epididymides (p = 0.09), the mean AUC0-24 was significantly higher in the non-infected left epididymis (p < 0.0001). The AUC0-24 and sparfloxacin concentrations of the right infected epididymis were significantly higher than those observed in the serum (p < 0.0001). In both testes, the AUC0-24 and sparfloxacin concentrations were lower than in the serum (p < 0.0001), however, the concentration exceeded the MIC tenfold for approximately 20 h. It is concluded that the pharmacokinetic properties of sparfloxacin (good in vitro activity, high penetration into the prostate gland, testes, infected and non-infected epididymides) make this drug a recommendable choice for the initial treatment of acute epididymitis caused by E. coli.

Antigenic and molecular analyses of different Chlamydia pneumoniae strains.

Chlamydia pneumoniae is an important human respiratory pathogen. Classification of C. pneumoniae isolates into distinguishable serovars or genotypes has not yet been reported. To determine whether antigenic or molecular variants among C. pneumoniae isolates exist, six strains were studied via immunoblot analysis and DNA sequence determination of the entire major outer membrane protein (MOMP) gene omp1. The strains included four prototype strains and two clinical isolates from our laboratory. Immunoblot analysis of sera from patients infected with C. pneumoniae revealed antigenic differences between the C. pneumoniae strains. Strong reactivity of one serum sample with a 65-kDa protein in two C. pneumoniae strains which was not observed with the other strains was the most prominent finding. All sera reacted with the 40-kDa MOMP. Comparison of the omp1 DNA sequences revealed that the omp1 genes of all strains were identical and were 100% identical to the sequence of the omp1 gene of C. pneumoniae AR-39. The results of this study demonstrate that unlike C. trachomatis, the omp1 gene is conserved in C. pneumoniae. Furthermore, it was shown that C. pneumoniae strains are antigenically different. This finding indicates that more than one serovar of C. pneumoniae exist.

Infection with Chlamydia pneumoniae in infants and children with acute lower respiratory tract disease.

The role of Chlamydia pneumoniae in the etiology of acute lower respiratory tract infections in infants and children is little understood. We studied the prevalence of C. pneumoniae infection in hospitalized infants and children with acute lower respiratory tract disease by cell culture, polymerase chain reaction (PCR), enzyme immunoassay and serology. Of 290 patients with a mean age of 3.7 years, only 3 (1%) were identified to be infected with C. pneumoniae. One child was positive in the cell culture as well as the PCR assay. Another infant was PCR-positive only and serologic evidence of infection was observed in a culture- and PCR-negative child. Chlamydia trachomatis was not detected in any patient specimen by either culture or PCR. Results of this study indicate that C. pneumoniae plays a minor role in the etiology of respiratory tract infections in infants and young children.

[Urethro-adnexitis in the man and acute urethral syndrome in the woman. Microbiological and immunologic studies of etiologic classification]. / Urethroadnexitis des Mannes und akutes Urethralsyndrom der Frau. Mikrobiologische und immunologische Untersuchungen zur ätiologischen Klassifikation.

Both common pathogens and unconventional, fastidious bacteria, viruses, parasites, and fungi are causative agents in male urethro-adnexitis and in female acute urethral syndrome. Uropathogens, Neisseria gonorrhoeae, Treponema pallidum, Mycobacterium tuberculosis, Chlamydia trachomatis, Mycoplasma spp., Haemophilus ducreyi, Calymmatobacterium granulomatis, Gardnerella vaginalis, anaerobic bacteria, Herpes simplex virus type II (HSV II), papillomaviruses (HPV), Trichomonas vaginalis and Candida spp. must be considered. The various diagnostic procedures and criteria applied for aetiological classification in cases of balanitis, urethritis, prostatitis, epididymitis, orchitis, and acute urethral syndrome are reviewed and evaluated.

Comparative efficacies of ofloxacin, cefotaxime, and doxycycline for treatment of experimental epididymitis due to Escherichia coli in rats.

The in vivo efficacy of ofloxacin was compared with those of cefotaxime and doxycycline in a rat model of epididymitis due to Escherichia coli. Treatment was started 24 h after infection and was continued for 7 days. Ofloxacin reduced the numbers of E. coli organisms in the epididymides significantly more than the other therapeutic regimens and cured the infection more frequently. Histopathological changes in the epididymides of ofloxacin-treated animals were significantly less severe than those observed in untreated animals. Doxycycline was less effective than ofloxacin but significantly reduced the titers of organisms in rat epididymides. In contrast, despite excellent in vitro activity, cefotaxime failed to reduce the magnitude of infection. The results of this study suggest that ofloxacin may be a very effective antimicrobial agent for the treatment of epididymitis due to E. coli.

Immuno-competent cells in the murine epididymis following infection with Escherichia coli.

Epididymitis was induced by retrograde injection of Escherichia coli into the vas deferens of 28 mice. A group of 28 saline-injected animals served as controls. On Days 1, 3, 7 and 28, groups of seven animals were killed. Bacterial culture was performed. Leucocyte numbers and distribution were determined in epididymides. In infected mice, E. coli were isolated from all epididymides on Days 1 and 3, but only from five of seven epididymides on Days 7 and 28. One week after infection, the total number of macrophages rose from about 10 to 28%. Significantly increased macrophage percentages were also found in animals killed 28 days after infection. A simultaneous increase in MHC class II positive cells was seen on Day 7. A total of 20% of the cells expressed MHC class II in infected epididymides (normal = 6%). A similar increase was found on Day 28 after infection. Most of the macrophages and MHC class II positive cells were located in the interstitium, fewer in the peritubular layer and nearly none in the epithelium. The main increase in these cells occurred in the interstitium and, to a lesser but significant extent, in the peritubular area. T-helper and T-suppressor/cytotoxic lymphocytes reached peak values on Day 28. The increase in T-lymphocytes and simultaneous appearance of plasma cells followed the increase in numbers of macrophages and MHC class II positive cells. They were located mainly in the interstitium. A sequential increase in leucocyte subsets and negative culture results for E. coli were observed on Days 7 and 28 (2/7 on each day). The inflammatory process was restricted to the interstitium.(ABSTRACT TRUNCATED AT 250 WORDS)

Acute lower respiratory tract infection associated with Chlamydia pneumoniae in Germany.

Serum specimens from 223 patients with acute lower respiratory tract infection were examined for antibodies to Chlamydia pneumoniae using the microimmunofluorescence test. Antibodies to Chlamydia pneumoniae were detected in 18 (20%) of 91 children and 64 (48%) of 132 adults. Among those individuals, 4 (4%) children and 15 (11%) adults had elevated IgG antibody titres indicating acute or recent infection. Specific IgM antibodies were observed in two patients. These results suggest that a significant proportion of lower respiratory tract infections in Germany is caused by Chlamydia pneumoniae.

Experimental epididymitis due to Chlamydia trachomatis in rats.

A new animal model of epididymitis due to Chlamydia trachomatis was developed. Adult male Wistar rats were inoculated in the vas deferens with C. trachomatis biovar mouse pneumonitis. After infection, C. trachomatis was recovered from the epididymides for up to 90 days. At day 30, organisms were also reisolated from the testes. Clinical findings included enlargement of infected epididymides and concurrent atrophy of the ipsilateral testes. Histological lesions in the epididymides consisted of pyogranulomatous inflammation, abscesses, and spermatic granulomas. Infection of the testis by C. trachomatis was associated with pyogranulomatous changes. In addition, testicular degeneration, characterized by moderate to severe loss of the germinal epithelium, was noted. Chlamydial antigen was detected within epithelial cells, intratubular macrophages, and macrophages in the stroma of the epididymis by immunoperoxidase staining. This rat model of chlamydial epididymitis appears to clinically and histopathologically mimic the human disease. This model offers the opportunity for further studies on the pathogenesis and sequelae of chlamydial epididymitis.

Semen parameters in men with and without proven chronic prostatitis.

In patients with complaints of chronic prostatitis, analysis of prostatic secretions for leukocytes and the use of the four-specimen technique allow a distinct classification of chronic bacterial (CBP), nonbacterial prostatitis (NBP), and prostatodynia (Pd). In this study, 32 men with CBP, 102 men with NBP, 142 men with Pd, and 42 volunteers, classified as mentioned above, underwent a two-fold ejaculate analysis using WHO criteria. Sperm count, progressive motility, range of abnormal spermatozoa, increased numbers of common bacteria, and peroxidase-positive leukocytes were analyzed. Additionally, antibody-coated bacteria (ACB), numbers of ureaplasmas in semen, and urethral colonization by chlamydia were investigated. Mean values of sperm density, motility, and morphology revealed no differences between the groups. Significant bacteriospermia (greater than or equal to 10(3) bacterial/ml) was evident in only 47% of the CBP group versus 6.8% (NBP), 16% (Pd), and 4.2% (controls). ACB was positive in 31 of 32 men with CBP versus 3 of 102 with NBP, 9 of 142 with Pd, and none of the controls. Increased numbers of leukocytes were evident in CBP and NBP patients compared to the controls (p less than or equal to 0.001) but were also present in patients of the NBP and Pd groups with chlamydial infections.

Recurrent haemospermia--underlying urogenital anomalies and efficacy of imaging procedures.

Recurrent haemospermia is often regarded as "essential". Following routine diagnostic procedures, we found an associated factor in 54 of 72 patients (75%). With one exception (prostatic carcinoma), the benign character of the disease was confirmed. Urogenital infection was the most frequent concomitant finding (50%); in 26 men chronic prostatis was diagnosed. Other disorders such as hypertension and coagulation disorders played a minor role. Additional investigation of the prostate gland and seminal vesicles by transrectal prostatic ultrasonography revealed persistent asymmetry of the latter glands as the main finding in 20 men (28%). In every case seminal vesicle carcinoma was excluded; haemorrhage due to cystic distension, inflammatory lesions or ductal obstruction was associated in all cases with congenital abnormalities, chronic urogenital infection, coagulation disorders or hypertension.