Chlamydia pneumoniae Induces Interferon Gamma Responses in Peripheral Blood Mononuclear Cells in Children with Allergic Asthma.

Respiratory infections caused by Chlamydia pneumoniae have been associated with exacerbations of asthma. Cell-mediated immunity (CMI) is critical for maintaining immunity. We compared interferon (IFN)-γ responses in C. pneumoniae-infected peripheral blood mononuclear cells (PBMC) in paediatric patients ± asthma. Presence of C. pneumoniae was tested from asthma patients (N = 17) and non-asthmatic controls (N = 16) (PCR). PBMC were infected for 1 h ± C. pneumoniae AR-39 (MOI = 0.1) and cultured for 48 h. IFN-γ levels were measured in supernatants (ELISA). C. pneumoniae-IgG antibodies in serum were determined (MIF). All subjects tested negative for C. pneumoniae (PCR). C. pneumoniae-induced IFN-γ production in vitro was more prevalent in asthma compared with non-asthma; levels of IFN-γ were higher in asthma compared with non-asthma (P = 0.003). There was no association between recent respiratory infection and positive IFN-γ responses. These data show that C. pneumoniae modulates IFN-γ responses in patients with asthma, even in absence of active infection.

Increased seroprevalence of Enterovirus 71 IgE antibodies in asthmatic compared with non-asthmatic children.

BACKGROUND: Asthma is a common pediatric chronic inflammatory airway disease. Respiratory viral infections are frequent infectious triggers for exacerbations of asthma. OBJECTIVE: We sought to determine whether Enterovirus 71 (EV71), a ubiquitous virus that causes systemic inflammatory responses in children but is not a known respiratory pathogen, can also serve as an infectious trigger for asthma. METHODS: Specific EV71 IgE and IgM antibodies (Abs), total serum IgE, and IL-2 and IL-4 cytokine levels in serum of asthmatic and non-asthmatic children (N = 42, ages 5-19; N = 35, ages 1-20, respectively) were measured (ELISA). RESULTS: Asthmatic children had higher EV71 IgE Ab levels than non-asthmatic (P < 0.001). Non-asthmatic children had significantly higher EV71 IgM Ab levels than asthmatic (P < 0.001). Despite low serum IgE levels of non-asthmatic, compared with asthmatic (P < 0.001), the non-asthmatic children produced significantly more IL-2 and IL-4 than asthmatic (P < 0.001; P < 0.001). The ages of the asthmatics, but not the non-asthmatics had a significant effect on the levels of EV 71 IgE Abs (P = 0.02; P = 0.356). A test of difference between these two slopes was significant. However, the ages of the non-asthmatic, but not the asthmatic children had a significant effect on the levels of EV 71 IgM Abs; a test of difference between these two slopes was significant. CONCLUSIONS: Increased specific EV71 IgE Ab responses may indicate that EV71 infection may also be an infectious trigger in asthma. However, the role of specific EV71 IgM Abs, Th2 cytokines, and age in non-asthmatic children should be further studied.

Chlamydia pneumoniae induces interleukin-12 responses in peripheral blood mononuclear cells in asthma and the role of toll like receptor 2 versus 4: a pilot study.

BACKGROUND: Chlamydia pneumoniae causes respiratory infection in adults and children, and has been associated with asthma exacerbations and induction of Immunoglobulin (Ig) E responses. We previously reported that C. pneumoniae enhances T helper (Th) 2 responses of peripheral blood mononuclear cells (PBMC) from asthmatic patients. It is likely that toll like receptor (TLR)-2 and TLR-4 mediate cytokine responses and host defense against C. pneumoniae. Thus, we sought to determine whether engagement of TLR-2 or TLR-4 may induce IL-12 production in our C. pneumoniae model. METHODS: PBMC (1.5 × 106) from asthmatic patients (N = 10) and non-asthmatic controls (N = 5) were infected or mock-infected for 1 h ± C. pneumoniae TW183 at a multiplicity of infection (MOI) = 1 and MOI = 0.1, and cultured for 48 h ± anti- TLR-2 and TLR-4 antibodies (Abs) (1 mg/mL). Interleukin (IL)-12 (48 h p.i.) and total IgE levels (day 10) were measured in supernatants (ELISA). RESULTS: High IgE levels were detected in supernatants of C. pneumoniae- infected PBMC from asthmatics on day 10, compared with mock-infected PBMC (p < 0.03). In contrast, IgE was not detected (<0.3 ng/mL) in either C. pneumoniae infected or mock-infected PBMC from non-asthmatics. IL-12 production by C. pneumoniae-infected asthmatic and non-asthmatic PBMC were similar. When anti-TLR4, but not anti-TLR2, was included in culture, IL-12 production by C. pneumoniae- infected asthmatic PBMC decreased. CONCLUSIONS: C. pneumoniae infection induces IgE production and modulates IL-12 responses in patients with asthma, which may be caused, in part, by differences in TLR-2 and TLR-4 stimulation.

Chlamydia trachomatis respiratory infection in Dutch infants.

Chlamydia trachomatis is the most common bacterial pathogen causing sexually transmitted infections in Dutch adults. As prenatal screening for C trachomatis and treatment of pregnant women is not routine practice in The Netherlands, perinatal transmission of C trachomatis may therefore occur. The presence of C trachomatis in infants less than 6 months of age who presented with respiratory complaints to the Erasmus MC-Sophia hospital was evaluated. Respiratory specimens, primarily nasopharyngeal swabs, were tested for C trachomatis, respiratory viruses and Mycoplasma pneumoniae using PCR, viral isolation in cell cultures and direct immunofluorescence. C trachomatis respiratory tract infection was confirmed to be relatively common with detection in 10 of 148 (7%) infants tested. C trachomatis had not been tested for by the attending physicians, but was the second most frequently detected respiratory pathogen after human Respiratory Syncitial Virus, which was found in 41 (28%) infants.

In vitro activity of a novel diaminopyrimidine compound, iclaprim, against Chlamydia trachomatis and C. pneumoniae.

The in vitro activities of iclaprim, a novel dihydrofolate reductase inhibitor, azithromycin, and levofloxacin were tested against 10 strains of Chlamydia trachomatis and 10 isolates of Chlamydia pneumoniae. For C. trachomatis and C. pneumoniae, the iclaprim MIC and minimal bactericidal concentration at which 90% of isolates were inhibited (MIC(90) and MBC(90)) were 0.5 micro g/ml, compared to an azithromycin MIC(90) and MBC(90) of 0.125 micro g/ml and levofloxacin MIC(90)s and MBC(90)s of 1 micro g/ml for C. trachomatis and 0.5 micro g/ml for C. pneumoniae.

Ultrastructural study of Chlamydia pneumoniae in a continuous-infection model.

We have established an in vitro model of long-term continuous Chlamydia pneumoniae infection in HEp-2 cells. Using transmission electron microscopy, we demonstrated the presence of spontaneous abnormal chlamydial inclusions similar in appearance to the persistent chlamydial forms induced in vitro by treatment with cytokines or antibiotics or by nutrient deprivation.

Activity of telithromycin, a new ketolide antibacterial, against atypical and intracellular respiratory tract pathogens.

Atypical respiratory pathogens such as Mycoplasma pneumoniae and intracellular pathogens such as Legionella spp. and Chlamydia spp. form a significant proportion of the aetiological agents underlying community-acquired pneumonia (CAP). The clinical signs or radiological features of atypical pneumonia are generally insufficient to predict accurately the pathogen involved; in addition, high costs and a considerable length of time are involved in the identification of atypical pathogens. Treatment is, therefore, most often empirical, and it is important that the activity of antibacterial agents available to treat CAP is sufficiently broad to eradicate infection with both common and atypical bacterial pathogens. Telithromycin (HMR 3647) is the first of a new family of antibacterials, the ketolides, and has been designed specifically for the treatment of community-acquired respiratory tract infections (RTIs). The excellent activity of telithromycin against the respiratory tract bacterial pathogens most commonly associated with community-acquired RTIs, including resistant strains, is well established. This review examines the considerable body of evidence showing that telithromycin also has a high level of activity against atypical and intracellular respiratory tract bacterial pathogens.

Standardizing Chlamydia pneumoniae assays: recommendations from the Centers for Disease Control and Prevention (USA) and the Laboratory Centre for Disease Control (Canada).

Chlamydia pneumoniae has been associated with atherosclerosis and several other chronic diseases, but reports from different laboratories are highly variable and "gold standards" are lacking, which has led to calls for more standardized approaches to diagnostic testing. Using leading researchers in the field, we reviewed the available approaches to serological testing, culture, DNA amplification, and tissue diagnostics to make specific recommendations. With regard to serological testing, only use of microimmunofluorescence is recommended, standardized definitions for "acute infection" and "past exposure" are proposed, and the use of single immunoglobulin (Ig) G titers for determining acute infection and IgA for determining chronic infection are discouraged. Confirmation of a positive culture result requires propagation of the isolate or confirmation by use of polymerase chain reaction (PCR). Four of 18 PCR assays described in published reports met the proposed validation criteria. More consistent use of control antibodies and tissues and improvement in skill at identifying staining artifacts are necessary to avoid false-positive results of immunohistochemical staining. These standards should be applied in future investigations and periodically modified as indicated.

Failure to demonstrate Chlamydia pneumoniae in cardiovascular tissue from children with Kawasaki disease.

Chlamydia pneumoniae has been associated with atherosclerosis and myocardial infarction in adults. Coronary artery tissues of five fatal cases of Kawasaki disease were examined by PCR; none was positive.

Mycoplasma pneumoniae infections.

Mycoplasma pneumoniae is a frequent cause of community-acquired respiratory infections in children and adults. Although the organism is felt to be the most frequent 'atypical' pathogen responsible for community-acquired pneumonia in adults, the prevalence of M. pneumoniae varies greatly from study to study, depending on the population and the diagnostic methods used. Recent studies have found the prevalence of M. pneumoniae in adults with pneumonia to range from 1.9 to over 30%. M. pneumoniae is also a frequent cause of outbreaks of respiratory disease in institutional settings. However, the diagnosis of M. pneumoniae infection is hampered by the lack of standardized, rapid, specific methods. This problem was illustrated by the results of an investigation of an outbreak of M. pneumoniae infection in a federal training facility. Accurate diagnosis required a combination of polymerase chain reaction and serology, as IgM antibodies were not present early in the course of the infection in many patients. Several papers evaluating various serological and polymerase chain reaction assays were published during the period of this review. An assessment of the actual performance of these tests was also hampered by the lack of standardized comparative methods. M. pneumoniae is susceptible in vitro to macrolides, tetracyclines and quinolone antibiotics; however, data are limited on the microbiological efficacy of these agents. Several pneumonia treatment studies were published during this period, practically all of them based the diagnosis of M. pneumoniae infection on serology; different methods and criteria were used in each study, and thus the microbiological efficacy could not be assessed. The Infectious Disease Society of America recently stated in their revised Practice Guidelines for the Management of Community-Acquired Pneumonia in Adults that, as there were no diagnostic tests available that reliably and rapidly detect M. pneumoniae, therapy must usually be empirical.

Is Chlamydia pneumoniae present in brain lesions of patients with multiple sclerosis?

We investigated the presence of Chlamydia pneumoniae in 81 normal and pathological specimens obtained from postmortem brain tissues of patients with multiple sclerosis and with other neurological or nonneurological diseases. The assays used included PCR amplification of all DNA samples in the initial study. Culture and a second PCR amplification of the organism in a subset of 19 brain specimens were also performed in two separate laboratories. All results were negative. Thus, this study on a large number of brain tissues suggests that C. pneumoniae is not involved in inflammatory demyelination.

Is there an association between Kawasaki disease and Chlamydia pneumoniae?

The clinical and epidemiological features of Kawasaki disease (KD) are consistent with an infectious cause. Because chronic infection with Chlamydia pneumoniae has been implicated in the pathogenesis of atherosclerosis, it has been suggested that it may also be involved in the pathogenesis of KD. Paired sera (baseline pretreatment and 1 year after treatment with intravenous immunoglobulin [IVIG]) from 26 children with KD and 29 age-matched controls were examined by microimmunofluorescence (MIF) serology and immunoblotting. There were no significant differences in the prevalence of anti-C. pneumoniae IgG, IgA, or IgM between cases and controls; however, 73%-85% of sera from cases and controls reacted with C. pneumoniae proteins by immunoblotting. There was significantly more reactivity in the pre-IVIG, but not post-IVIG, KD sera compared with sera from controls to proteins at 72-74 kDa and 74-76 kDa. They may be heat shock proteins. The results of this study do not support an association between KD and C. pneumoniae on the basis of MIF and immunoblot analysis.

Microbiologic efficacy of moxifloxacin for the treatment of community-acquired pneumonia due to Chlamydia pneumoniae.

Nasopharyngeal specimens for culture of Chlamydia pneumoniae were obtained from patients participating in two pneumonia treatment studies: an open study of 400 mg moxifloxacin orally, qds for 10 days and a randomized, double-blind comparison of moxifloxacin, 400 mg orally, qds versus clarithromycin, 500 mg orally, bd, both for 10 days. C. pneumoniae was eradicated from the nasopharynx of seven of ten (70%) microbiologically evaluable patients who were treated with moxifloxacin and four of four who were treated with clarithromycin. Minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) of 21 isolates of C. pneumoniae from 18 patients obtained before and after therapy were performed against moxifloxacin and clarithromycin. The MIC(90)s and MBC(90)s for moxifloxacin and clarithromycin were 1 and 0.06 mg/l, respectively. The MICs and MBCs against moxifloxacin of six isolates from three persistently infected patients who were treated with the drug were the same at baseline and follow-up. The persistence of C. pneumoniae after treatment with moxifloxacin was probably not due to the emergence of resistance.

Activity of gemifloxacin and other new quinolones against Chlamydia pneumoniae: a review.

Quinolones are currently used as empirical therapy for treatment of community-acquired lower respiratory infections as they are effective against a broad range of conventional bacterial and 'atypical' pathogens, including Chlamydia pneumoniae. C. pneumoniae is estimated to be associated with 10-20% of community-acquired pneumonia in adults, and has recently been suggested to play a role in several non-respiratory conditions, including atherosclerosis. The newer, third-generation quinolones have enhanced activity against Gram-positive bacteria, including Streptococcus pneumoniae, and prolonged serum half-lives that permit once-daily dosing. Although gemifloxacin (SB-265805) and other new quinolones have good activity against C. pneumoniae in vitro, practically all published treatment studies have relied on serological diagnosis. Consequently, the microbiological efficacy of these agents in human infection has not been assessed. This paper reviews what is known to date of the in vivo microbiological efficacy of the quinolones against C. pneumoniae, and demonstrates the importance of assessing this parameter when evaluating the clinical utility of these agents in C. pneumoniae infection.