Detection of serum antibodies to Bartonella henselae and Coxiella burnetii from Japanese children and pregnant women.

The participation of Bartonella henselae and Coxiella burnetii in the pathogenesis of fever of unknown origin (FUO) and lymphadenopathy has not been completely clarified. Prevalence of these two agents in Japanese children is also unknown. Serum IgG and IgM antibodies to B. henselae and to C. burnetii were examined by the indirect fluorescence antibody assay. Enzyme immunoassay kits were used to detect serum IgG and IgA antibodies against Chlamydia trachomatis. Out of 200 healthy normal pregnant women, two (1.0%) had serum IgG antibodies to B. henselae, four (2.0%) to C. burnetii and 49 (24.5%) to C. trachomatis. Out of 29 patients with FUO, one (3.4%) had serum IgG antibodies to B. henselae, four (13.8%) to C. burnetii and none to C. trachomatis. Out of 31 patients with cervical lymphadenopathy, three (9.6%) had serum IgG antibodies to B. henselae, two (6.5%) to C. burnetii and none to C. trachomatis. Out of 22 patients with generalized lymphadenopathy, one (4.5%) had serum IgG antibodies to B. henselae, three (13.6%) to C. burnetii and none to C. trachomatis. Prevalences of serum antibodies to C. burnetii in the patients with FUO and generalized lymphadenopathy and to B. henselae in the patients with cervical lymphadenopathy were significantly higher than those of normal pregnant women (Welch's t-test; P<0.01). These two agents may have some roles in the pathogenesis of FUO and lymphadenopathy in Japanese children.

Therapeutic effect of clarithromycin for respiratory-tract infections in children caused by Chlamydia pneumoniae. Research Group of Sapporo for Pediatric Chlamydial Infections.

Children infected with Chlamydia pneumoniae sometimes experience lower respiratory tract infections such as pneumonia and bronchitis. Although numerous anti-microbial compounds have been reported to be active against the organism, most of them have not been in a clinical trial in infants and children with C. pneumoniae infection. Clarithromycin has been shown to express anti-chlamydial effects in vitro. In this study, we evaluated the clinical anti-C. pneumoniae properties of clarithromycin in children with mainly lower respiratory tract infection. We administered clarithromycin orally to 21 infants and children at a dose of 10-15 mg/kg/day divided into two or three doses for 4-21 days. Clinical symptoms, roentgenographic and laboratory abnormal findings improved. The overall clinical efficacy rate was 85.7% (18 of 21 cases). Administration of clarithromycin was considered to be a suitable treatment for improving lower respiratory infections in infants and children caused by C. pneumoniae.

Analysis of Chlamydia trachomatis serovars in endocervical specimens derived from pregnant Japanese women.

The polymerase chain reaction (PCR) method has been employed to amplify a chlamydial genome encoding four variable segments of the major outer membrane protein and genotyping of different Chlamydia trachomatis serovars was successfully achieved by means of restriction fragment length polymorphism (RFLP) analysis and sequencing of amplified DNA. These methods were applied to identify the serotypes of C. trachomatis in endocervical specimens obtained from asymptomatic pregnant Japanese women at 28-30 weeks of gestation. Among the 218 specimens, 207 were serotyped 43 (19.3%) as serovar D, 53 (24.3%) as E, 24 (11.0%) as F, 39 (17.9%) as G, 15 (6. 9%) as H, 15 (6.9%) as I, five (2.3%) as J, nine (4.1%) as K and four (1.8%) as mixed. Among the 11 unclassified strains by RFLP, six (2.8%) were identified as serovar B variants and five (2.3%) were identified as D/IC-Cal-8. It was suggested that variants of endemic trachoma serovars also have affinity for the urogenital tract of Japanese pregnant women.

Subtyping of cytomegalovirus strains obtained from immunocompetent children.

We analyzed 33 clinical isolates of cytomegalovirus (CMV) obtained from immunocompetent Japanese children. DNA was extracted from infected MRC-5 cells and we amplified CMV glycoprotein-B (gB), major IE (MIE), DNA polymerase and glycoprotein-H (gH) genes using PCR. Among the 33 clinical isolates, 24 were identified as gB group 1, one as group 2, 6 as group 3, and 2 as simultaneous infections with group 1 and group 3 strains. Clinical isolates of CMV have also been classified into 18 MIE, 8 DNA polymerase and 15 gH genotype patterns by restriction fragment length polymorphism (RFLP). RFLP analysis is useful in molecular epidemiological studies of CMV infection in immunocompetent individuals.

Relationship between seropositivity of husbands and primary cytomegalovirus infection during pregnancy.

The role of the sexual transmission of human cytomegalovirus (CMV) as a cause of congenital infection was investigated. Serum samples were collected from 756 pregnant women at 10 to 12 weeks of gestation and at 32 to 36 weeks of gestation. Serum samples were also obtained from the husbands of women who seroconverted and women who were seronegative during pregnancy. Commercially available enzyme immunoassay kits were used to detect serum IgG, IgM, and IgA antibodies against CMV. CMV from neonatal urinary specimens was isolated according to a standard tissue culture technique, using MRC-5 cells. At 10 to 12 weeks of gestation, 634 of the 756 pregnant women (83.9%) had IgG antibody to CMV. At 32 to 36 weeks of gestation, 642 of the 756 women (84.9%) had IgG antibody to CMV. A meaningful rise of serum IgG-antibody titer (seroconversion) occurred in 8 women (1.1%). CMV was isolated from the urine of an infant born to a seroconverted woman within a week after birth. The prevalence of IgG antibody to CMV was significantly higher in the husbands of women who seroconverted during pregnancy than in the husbands of the women who were seronegative during pregnancy (P < 0.01). Understanding the epidemiology of CMV is a key element in the development of strategies for the prevention of infection. The transmission of CMV by sexual contact may be important in the pathogenesis of congenital infection. Entirely new approaches to the prevention and treatment of congenital CMV infection are necessary, including antiviral interventions and the development of a vaccine strategy.

Analysis of immune cells in a patient with post-transfusion hepatitis caused by human cytomegalovirus.

Cellular immune responses are associated with the pathogenesis of human cytomegalovirus (HCMV) hepatitis. We investigated a patient with post-transfusion HCMV hepatitis. A 9 year-old girl was involved in a traffic accident and suffered from traumatic damage to the left kidney and diaphragm and received a pelvic bone fracture. At emergency surgery she was transfused with 1200 ml of fresh whole donor blood. Abnormal liver function was observed in the 10 days after surgery. Titers of serum anti-HCMV IgG and IgM antibodies were elevated at 11, 17 and 25 weeks after operation. We analyzed the surface markers of peripheral blood mononuclear cells obtained 21 weeks after surgery. The CD4/CD8 ratio and the number of CD16 + CD56 decreased. We detected HCMV immediate early (IE) DNA in the fractionated peripheral blood cells (polymorphonuclear leukocytes, CD2+, CD4+ and CD8+ T lymphocytes) by polymerase chain reaction. The histology of liver biopsy at 23 weeks after operation showed the findings of acute hepatitis and the absence of HCMV IE antigen. It was considered that the immunosuppressive condition associated with the trauma, operation or transfusion itself induced the reactivation of HCMV or that transfused blood cells infected with HCMV caused reinfection. It was also speculated that HCMV hepatitis was not only due to the direct damage of hepatic cells by HCMV, but also due to the cellular immune responses associated with HCMV infection.

Evaluation of serological tests for screening of chlamydial eye diseases.

Trachoma is recognized as one of most important origins of blindness in developing countries and inclusion conjunctivitis is associated with STD in developed countries. We evaluated the diagnostic value of serological tests for the screening of eye diseases associated with Chlamydia trachomatis infection. We determined serum IgG, IgA and IgM antibodies to C. trachomatis from 53 Japanese patients with active inflammatory trachoma (aged more than 60 years) and from 107 adult patients (aged 20 to 50 years) with acute inclusion conjunctivitis by ELISA test kit. We detected serum IgG antibodies from 22 out of 53 (42.5%) patients with trachoma and from 40 out of 107 (37.4%) patients with acute inclusion conjunctivitis. We also detected serum IgM antibodies from 7 out of 53 (13.2%) patients with trachoma and from 35 out of 107 (32.7%) patients with acute inclusion conjunctivitis. The prevalence of serum IgM antibodies to C. trachomatis in patients with acute inclusion conjunctivitis was significantly higher than that in patients with active trachoma (p < 0.05). Serological tests are also thought to be useful for screening of chlamydial eye diseases.

Inhibitory effect of povidone-iodine for the antigen expression of human cytomegalovirus.

Anti-human cytomegalovirus (HCMV) properties of povidone-iodine (PVP-I) were evaluated in vitro. The effect of PVP-I was evaluated by indirect immunofluorescence assay on HCMV-infected MRC-5 cells. Percentages of fluorescent cells positive for HCMV immediate early and early antigens in cultures inoculated with AD 169 treated with PVP-I at various concentrations and reaction times were compared with the number of fluorescent cells in the controls inoculated with the virus alone. PVP-I was found to exert some inhibitory effect at a concentration of 0.5% and complete inhibition at a concentration of 7.5% on the infection of MRC-5 cells by HCMV AD 169 strain. Entirely new approaches to the prevention of HCMV infections in infants are necessary. The adequate use of PVP-I as a disinfectant may reduce the transmission of HCMV.