Biomedical subjects

C C Kuo

Publications and source records attributed to C C Kuo.

At least 181 records · Page 10Linked to original sources

PubMed1988

In vitro growth of Chlamydia trachomatis in conjunctival and corneal epithelium.

This study used primary cultures of conjunctival and corneal epithelial cells from rabbit, monkey and humans to investigate the infection process and host interactions of chlamydia. The epithelial cells were isolated from bulbar and palpebral conjunctivae and the cornea following incubation with EDTA or dispase and microdissection. The cells were trypsinized and grown in microtest wells or Rose Chambers (gelatin-coated glass substratum). The cell origin of the cultured epithelial cells was verified by immunofluorescence staining with anticytokeratins. The cells were nonreactive when stained with antivimentin, a fibroblast marker. The growth kinetics of the cultured epithelial cells were determined by morphological criteria. Cells reached confluency by 5-7 days, and remained as such for 18 days. The cells were maintained as long as 32 days in primary culture. After the cells had attained confluency (5-7 days), they were infected with ocular strains B/TW-5/OT or C/TW-3/OT C. trachomatis. Chlamydial inclusions were identified by light and electron microscopy. Direct FITC C. trachomatis staining (species-specific monoclonal antibody) of the inclusions was used to determine the infectivity of the various types of epithelial cells. The dynamics of the infection process was documented by time lapse photomicroscopy. Small inclusions were identified by 18 hr postinoculation (pi). By 48 hr pi, the inclusions had dramatically increased in size, occupying much of the host perikarya. The reticulate bodies were very active at this time. By 70 to 100 hr pi, the inclusions contained the highly infectious elementary bodies. At this time, rupture of the inclusion and lysis of the infected cells usually occurred.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1987

Differential antimicrobial activity of human mononuclear phagocytes against the human biovars of Chlamydia trachomatis.

The antimicrobial activities of human mononuclear phagocytes against Chlamydia trachomatis were investigated. Phagocytes cultured for 7 days or less were efficiently microbicidal. Almost complete inactivation of organisms from both human biovars was observed after 48 hr of incubation. However, organisms from the lymphogranuloma venereum (LGV) biovar survived in mononuclear phagocytes infected after 8 days or more in culture, whereas those from the trachoma biovar continued to be killed by such cells. Phagocytes cultured as long as 21 days killed the trachoma organisms with the same effectiveness as those cultured for 7 days or less. An ultrastructural study of inoculated phagocytes illustrated phagolysosomal fusion with degradation of organisms from either biovar in phagocytes which had been cultured for 24 hr before infection. Phagolysosomal fusion was not observed in cells which had been cultured for 8 days or more and then infected with LGV. The addition of interferon-gamma to these macrophages partially restored the phagocytes' microbicidal activity for LGV. Furthermore, a synergistic effect was observed when eosinophil peroxidase was added with interferon. Specific antibody failed to neutralize the infectivity of LGV organisms in 8-day or older mononuclear phagocytes. The findings may reflect the differences in disease syndromes between the two biovars, with the trachoma biovar causing more peripheral diseases and the LGV biovar causing a more systemic disease, with lymph node involvement as its main syndrome.

Blood Bactericidal Activity↗

PubMed1987

Identification of conserved regions for species and subspecies specific epitopes on the major outer membrane protein of Chlamydia trachomatis.

Proteolytic peptides containing the serological epitopes present on the major outer membrane protein of Chlamydia trachomatis were studied by immunoblots. Monoclonal antibodies which have been defined by micro-immunofluorescence typing as serovar-, subspecies and species-specific were utilized. The reactivity of either serovar-specific or species-specific monoclonal antibodies in the immunoblots was similar to that in the micro-immunofluorescence test. However, monoclonal antibodies which demonstrated subspecies-specific reactivity by the micro-immunofluorescence test expressed species-specific reactivity by the immunoblot method. The serovar-specific epitope which was identified on the major outer membrane protein was lost after proteolytic digestion with Staphylococcus aureus V8 protease. Immunoblots of the proteolytic fragments identified the species-specific epitope on several peptides and the subspecies-specific epitope on fewer peptides than those in which the species-specific epitope was identified. In addition, both the species and subspecies epitopes were present on the same fragments of different serovars. The data indicate that the species epitope is on conserved regions of the major outer membrane protein and is structurally clustered with a subspecies-specific epitope.

Animals↗

PubMed1987

Chlamydial infection of subcutaneous fimbrial transplants in cynomolgus and rhesus monkeys.

Acute infection with Chlamydia trachomatis serotype E was established in monkey fallopian tube fimbriae by subcutaneous implantation. Depending upon monkey species, from eight to 20 implants could be established in each animal. Animals were given estrogen before percutaneous inoculation of the autografts with Chlamydia. Acute inflammatory changes were found in homografts examined in the first week after infection, with chronic inflammatory changes noted thereafter. Chlamydial inclusions were detected within fimbrial epithelial cells up to seven days postinoculation by fluorescent-antibody staining and immunoperoxidase staining with C. trachomatis-specific monoclonal antibody. Organisms were recovered from autografts up to five days after infection. Analysis of serum antibody by microimmunofluorescence revealed that serotype E-specific antibody of both IgM and IgG classes was produced after infection. We conclude that subcutaneously implanted fallopian tube autografts may provide a useful primate model for kinetic studies of chlamydial infection and immunity.

Animals↗

PubMed1987

Distal tubal obstruction induced by repeated Chlamydia trachomatis salpingeal infections in pig-tailed macaques.

The effects of repeated infections with Chlamydia trachomatis, serovars F, D, and J, were examined in pig-tailed macaques. The fallopian tubes of three experimental monkeys were inoculated at the middle of the menstrual cycle on three consecutive months. Monkey 1 received homologous F; monkey 2 received heterologous F, D, and J; and monkey 3 received homologous inoculations in the right and heterologous inoculations in the left fallopian tubes. One control monkey (4) received repeated inoculations of HeLa cell materials only and remained normal throughout the experiment. Infection was confirmed by isolating the microorganism from both the endosalpinx and endocervix. Antibody to the infecting strains was demonstrated in sera, tears, and cervical secretions by using microimmunofluorescence. Mild chronic salpingitis developed in monkeys 1 and 3, and chronic follicular salpingitis developed in monkey 2. Peritubal and periadnexal scarring and endosalpingeal adhesion formation were produced after reinfection. The right fallopian tube of monkey 3 was distally obstructed (confirmed by hysterosalpingography). Systemic complications, including perihepatitis and conjunctivitis, were also documented in these monkeys. We conclude that repeated infections produced extensive tubal scarring, chronic salpingitis, and distal tubal obstruction, findings not apparent in primary infection.

Animals↗

PubMed1987

Delayed appearance of Chlamydia trachomatis infections acquired at birth.

To determine the delay in detectability of Chlamydia trachomatis infections acquired at birth, we serially evaluated 120 infants born vaginally to infected women. Specimens for isolation of Chlamydia were taken from several anatomic sites. Results were analyzed for the age and site of the initial positive culture from each infant. Of 112 infants tested in the first month of life, 22% were culture-positive in the conjunctiva and 25% were positive in the pharynx. Initial positive rectal and vaginal cultures were obtained only in the third and fourth months of life, and all initial vaginal cultures were associated with positive rectal cultures. The latency of C. trachomatis in infants exposed at birth is often more than 1 month and can be longer than 97 days. This latency might be caused by suppression of the growth of the organism by antibodies acquired in utero.

Antibody Formation↗

PubMed1987

Ultrastructural study of Chlamydia trachomatis surface antigens by immunogold staining with monoclonal antibodies.

Surface antigens of Chlamydia trachomatis were studied by immunogold staining with monoclonal antibodies and by electron microscopy. The serovar- and subspecies-specific epitopes were the most surface accessible. The species- and genus-specific epitopes were the least surface exposed. Similar serological specificity as that in the microimmunofluorescence test was demonstrated by immunogold staining.

Antibodies, Monoclonal↗

PubMed1987

Unique ultrastructure in the elementary body of Chlamydia sp. strain TWAR.

The ultrastructure of two prototype strains (TW-183 and AR-39) of Chlamydia sp. strain TWAR was described. The TWAR elementary body (EB) demonstrated a unique morphology and structure distinct from those of other chlamydial organisms. It was pleomorphic but typically pear shaped. The average size was 0.38 micron, with a long axis of 0.44 micron, a short axis of 0.31 micron, and a ratio of the long to the short axes of 1.42. The cytoplasmic mass was round, with an average diameter of 0.24 micron. There was a large periplasmic space. Small, round electron-dense bodies (0.05 micron in diameter), which were attached to the cytoplasm by a stringlike structure, were seen in the periplasmic space. These features are in contrast to those of other chlamydiae, which are typically round with a narrow or barely discernible periplasmic space. The TWAR reticulate body (RB) was morphologically and structurally similar to those of other Chlamydia species, having an average diameter of 0.51 micron and being circular in shape. The ultrastructural observations of the intracellular growth of TWAR in HeLa cells revealed that TWAR underwent the same developmental cycle as do other chlamydiae, i.e., transformation of EB to RB, multiplication by binary fission, and maturation by transformation of RB to EB via the intermediate-form stage.

Antigens, Bacterial↗

PubMed1987

Characterization of the new Chlamydia agent, TWAR, as a unique organism by restriction endonuclease analysis and DNA-DNA hybridization.

Several molecular techniques were used for comparison of the novel Chlamydia agent, TWAR, with Chlamydia trachomatis and Chlamydia psittaci. Unlike all serotypes of C. trachomatis and most strains of C. psittaci, the eight TWAR isolates examined did not contain extrachromosomal DNA. TWAR was readily distinguished from C. trachomatis or C. psittaci by restriction endonuclease analysis, whereas identical or nearly identical restriction patterns were observed among the TWAR isolates. Southern blot analysis with a gene encoding a portion of the C. trachomatis serovar L2 major outer membrane protein as the probe showed that TWAR, like C. psittaci, contained sequences homologous to this gene. However, while the hybridization patterns were identical for all TWAR isolates, they differed from those of any of the other Chlamydia species tested. A PstI gene bank containing TWAR DNA was constructed in pUC19. Random fragments were purified and used for probing Chlamydia chromosomal digests. All of the five probes tested were TWAR specific, with the TWAR isolates showing identical patterns of homology. Qualitative studies of the DNA homology revealed that TWAR did not have significant homology to any of the Chlamydia strains assayed. Collectively, these results demonstrate that the TWAR isolates represent a single strain or closely allied genotypes and are clearly distinct from any of the other chlamydiae tested.

Animals↗

PubMed1987

Pneumonia associated with the TWAR strain of Chlamydia.

From November 1981 to August 1984, 301 adult patients with community-acquired pneumonia were admitted to the major referral hospital of Nova Scotia. Serologic tests done on these patients included microimmunofluorescence using the TWAR strain of Chlamydia and all Chlamydia trachomatis serovars as antigens. The TWAR strain has been shown to cause mild pneumonia in teenagers and young adults. Of the 301 patients, 18 (6%) had serologic evidence of recent infection with the TWAR organism. Their mean age was 64 years. Pneumonia associated with the presence of acute TWAR antibody had no characteristic clinical or radiographic features when compared with pneumonia without acute chlamydia antibody. Six patients, who all had preexisting serious chronic disease, had severe illness, and 2 died. Both patients who died had complicated hospital courses and other concomitant infections. We conclude that the TWAR organism may cause pneumonia in older adults and persons with chronic diseases that require hospitalization.

Adolescent↗

PubMed1987

Experimental trachoma in subcutaneous conjunctival autografts in macaques.

To study chlamydial conjunctivitis, conjunctival autografts subcutaneously implanted in pockets on the abdomens of rhesus (Macaca mulatta) and pig-tailed (M. nemestrina) monkeys were inoculated percutaneously (4-10 X 10(4) inclusion-forming units per pocket) with trachoma strains of Chlamydia trachomatis (serovars B & C). These conjunctival pockets were removed on days 2, 3, 4, 5, 6, 7, 8, 10, 14 and 16 post-inoculation (pi). Tissues (at least two samples at each time interval) were prepared either for reisolation of the organisms by cell culture or histologic examination by light and electron microscopy. Control tissue, inoculated with either HeLa cell material or UV-inactivated organisms, were prepared in parallel. Bloods were drawn and tear strips taken at weekly intervals for 8 weeks. A total of 221 bulbar and 99 palpebral conjunctival pockets were established over time with the success rate of 75% and 88%, respectively, in six rhesus and ten pig-tailed monkeys. Histological examination revealed widespread infiltration of mixed polymorphonuclear and mononuclear cells on days 2 and 3 pi. By day 5 pi, lymphocytes had migrated into the thinned epithelial layer. Patches of inflammatory infiltrate similar to trachoma follicles were observed, although distinct germinal centers were absent. Surface morphologies of normal and infected pocket conjunctiva were examined by scanning electron microscopy. Normal tissue was characterized by a regular mosaic pattern of closely packed epithelial cells containing numerous microvilli. Infected tissue was edematous, and the continuity of the mucosal surface had been altered. Chlamydiae were reisolated from the pockets on days 2, 3, 6, 8 and 10 pi.(ABSTRACT TRUNCATED AT 250 WORDS)

Abdomen↗

PubMed1986

A new Chlamydia psittaci strain, TWAR, isolated in acute respiratory tract infections.

During a 2 1/2-year period, we studied 386 University of Washington students with acute respiratory disease, to determine whether a Chlamydia psittaci strain, here designated TWAR, is an important respiratory pathogen. Serologic evidence of recent TWAR infection was found in 13 students, and the organism was isolated from 8 of these. TWAR infection occurred in 12 percent of the students who had pneumonia (9 of 76), 5 percent of those with bronchitis (3 of 63), and 1 percent of those with pharyngitis (1 of 150). The TWAR infections occurred throughout the study period. Pharyngitis, often accompanied by laryngitis, was a common first symptom. Clinically, the infections resembled those with Myco-plasma pneumoniae; therefore, the patients were given courses of erythromycin used for the treatment of M. pneumoniae infections. This therapy proved to be inadequate. The limited data available suggest that the TWAR strain is a "human" C. psittaci that is spread from human to human, without a bird or animal host.

Acute Disease↗

PubMed1986

Etiology of cervical inflammation.

We studied the relationships of selected microbial, clinical, demographic, and behavioral variables to mucopurulent cervicitis in two clinical settings, a sexually transmitted disease clinic and a student health clinic. From each clinic, we studied a group of women referred for suspected mucopurulent cervicitis and a representative sample of other women attending the clinic. After the women were stratified by patient group and summary odds ratios for all groups were obtained, mucopurulent cervicitis was most strongly associated with the isolation of Chlamydia trachomatis; other variables associated with mucopurulent cervicitis included the isolation of Ureaplasma urealyticum, Gardnerella vaginalis, and Trichomonas vaginalis, the presence of serum antibody to C. trachomatis, the clinical diagnosis of bacterial vaginosis, and oral contraceptive use (positive associations) or isolation of yeast (negative association). After adjustment for cervical culture results for C. trachomatis, mucopurulent cervicitis was positively associated with oral contraceptive use (p = 0.02) and isolation of U. urealyticum (p = 0.02) and negatively associated with isolation of yeast (p = 0.03). Among women with a positive cervical culture for C. trachomatis, isolation of U. urealyticum was significantly associated with mucopurulent cervicitis, while among the subgroup of women with a negative cervical culture for C. trachomatis and positive serum antibody to C. trachomatis, oral contraceptive use was strongly associated with mucopurulent cervicitis. These results confirm that in both clinical settings C. trachomatis is the major cause of mucopurulent cervicitis. The roles of U. urealyticum, T. vaginalis, G. vaginalis, bacterial vaginosis, and oral contraceptive use in the etiology of mucopurulent cervicitis deserve further study.

Adult↗

PubMed1986

Degradation of Chlamydia trachomatis in human polymorphonuclear leukocytes: an ultrastructural study of peroxidase-positive phagolysosomes.

We have previously shown that human polymorphonuclear leukocytes (PMNs) killed organisms belonging to both human biovars of Chlamydia trachomatis. However, the mechanism of destruction was still unclear. We therefore conducted an ultrastructural and cytochemical study to investigate the mechanism of chlamydial degradation. PMNs were inoculated with the trachoma serovar B (B/TW-5/OT) or with the lymphogranuloma venereum serovar L2 (L2/434/Bu) for 15, 30, 60, or 120 min and then fixed and processed for transmission electron microscopy. Diaminobenzidine, a cytochemical marker, was used to demonstrate the localization of intracellular peroxidase. Ultrastructural evidence is presented showing the progressive degradation of chlamydiae over a 2-h period within peroxidase-positive phagolysosomes. Pretreatment of organisms with normal or immune serum was not required for the process of degradation.

Chlamydia trachomatis↗

PubMed1986

Identification of a new group of Chlamydia psittaci strains called TWAR.

A new group of Chlamydia psittaci strains has been identified. They are called TWAR after the laboratory designation of the first two isolates. Twelve strains were isolated from pharyngeal swabs of different persons with acute respiratory disease in Seattle, Wash., during 1983 to 1986. One strain was obtained from the eye of a child during the trachoma vaccine study in Taiwan in 1965. Nine strains were characterized in this study. TWAR organisms formed intracytoplasmic inclusions in HeLa cells which were morphologically typical of C. psittaci and iodine stain negative (contained no glycogen). Immunological analysis with various chlamydia-specific monoclonal antibodies revealed that TWAR strains belong to the genus Chlamydia, are distinct from C. trachomatis, and are serologically unique among C. psittaci. All TWAR strains so far isolated appear identical serologically. TWAR organisms grew poorly in egg and cell cultures and demonstrated low virulence to mice by intracerebral, intranasal, and intravenous inoculation. Available data suggest that the TWAR strain is a primary human pathogen.

Animals↗

PubMed1986

Pulmonic regurgitation and reconstruction of right ventricular outflow tract with patch. An experimental study.

Twenty-five dogs were divided into three experimental groups. In each animal of Group I (10 dogs), one pulmonary valve leaflet was resected and a monocusp-bearing bovine pericardial patch was sewn into the right ventricular outflow tract. The monocusp was retracted to the patch with plicating sutures, and when the sutures were removed the monocusp became functional. In each animal of Group II (10 dogs), an incision was made in the right ventricular outflow tract along the anterior pulmonary valvular commissure and a bovine pericardial patch of adjustable width (one half, one, or one and one half times the diameter of the pulmonary valve ring) was sewn into the incision. Group III (five dogs) was the control group. During each situation of altered pulmonary valve function, in each experimental animal, the pulmonary blood flow and intracardiac pressures were measured. Pulmonary regurgitation was expressed regurgitant fraction (retrograde flow divided by net forward flow). We reached the following conclusions: In all situations where pulmonary incompetence was created, the regurgitant fraction was related directly to the width of the patch and was usually not more than half of the net forward flow when the width of the patch was less than one and a half times the pulmonary valve ring diameter. Regurgitant fraction was about half of net forward flow when one leaflet had been excised. The index of pulmonary artery diastolic pressure minus right ventricular end-diastolic pressure was sensitive and bore an inverse relationship to the degree of pulmonary regurgitation. An accurately placed monocusp-bearing patch restored pulmonary valve competence after excision of a cusp. In treating patients, we are encouraged to preserve as much pulmonary valve function as possible.

Animals↗

PubMed1985

Prevalence and manifestations of endometritis among women with cervicitis.

Thirty-five women referred from a clinic treating sexually transmitted diseases, because of suspected cervicitis, were studied for the presence of endometritis by transcervical endometrial biopsies and cervical and endometrial cultures. Fourteen (40%) of the patients had histologic evidence of endometritis. Findings that significantly correlated with endometritis included a history of intermenstrual vaginal bleeding, the presence of Chlamydia trachomatis, Neisseria gonorrhoeae, or Streptococcus agalactiae in the cervix, and the presence of serum antibodies to C. trachomatis or to Mycoplasma hominis.

Antibodies, Bacterial↗

PubMed1985

Cytologic manifestations of cervical and vaginal infections. I. Epithelial and inflammatory cellular changes.

We used multiple logistic regression to analyze microbiological and clinical correlates of inflammatory and epithelial cell changes on Papanicolaou-stained cervical smears in patients from a sexually transmitted disease clinic. Among randomly selected patients, increased numbers of histiocytes and polymorphonuclear leukocytes and the presence of transformed lymphocytes were associated with Chlamydia trachomatis infection, while increased lymphocytes were associated with Trichomonas vaginalis infection; minimal squamous atypia was associated with yeast infection; and moderate squamous atypia and koilocytosis were associated with cervical condylomata visualized by colposcopy. Among patients referred for cervicitis, C trachomatis infection was also associated with reactive or atypical metaplastic cells. Distinct inclusions were seen by Papanicolaou smear in only 22% of C trachomatis infections. In randomly selected patients, however, the presence of transformed lymphocytes or increased histiocytes had a sensitivity of 95%, a specificity of 75%, and a positive predictive value of 50% in relation to isolation of C trachomatis, and could therefore be used for selection of patients for confirmatory testing for C trachomatis infection.

Adult↗