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Biomedical subjects

C C Kuo

Publications and source records attributed to C C Kuo.

At least 217 records · Page 12Linked to original sources
PubMed1983

Host response to primary Chlamydia trachomatis infection of the fallopian tube in pig-tailed monkeys.

Experimental acute salpingitis was produced in four pig-tailed monkeys, Macaca nemestrina, by intratubal inoculation with Chlamydia trachomatis (serotypes E or F). The organisms were reisolated from both the endosalpinx and endocervix as early as 1 week after the original inoculation. Endosalpinx cellular responses to the infection were examined by light, transmission, and scanning electron microscopy. A moderate lymphocyte infiltration was detected in the submucosa on day 7. By days 14 and 21 the lymphocytic infiltration was heavy and extended into both the submucosa and the mucosa; the infiltration subsided by day 35. Epithelial cell degeneration occurred in close approximation to lymphocytes, suggesting the immunologic basis of tissue destruction. Scanning electron microscopy revealed extensive deciliation and increased plasmalemmal alterations of nonciliated cells. The presence of C. trachomatis in frozen and deparaffinized tissues was demonstrated by immunofluorescence and immunoperoxidase staining with monoclonal antibody to C. trachomatis. Only secretory cells contained chlamydial inclusions. A humoral immune response to C. trachomatis was demonstrated by microimmunofluorescence. No histologic or immunologic evidence of infection was present in two control monkeys inoculated with HeLa cell material. The histopathologic and immunologic findings of this study establish the pig-tailed monkey as a useful model for further studies of the pathogenesis and pathophysiology of chlamydial salpingitis.

Acute Disease↗
PubMed1982

Increased frequency of serum antibodies to Chlamydia trachomatis in infertility due to distal tubal disease.

186 infertile women underwent standard infertility investigations (including hysterosalpingography and, in 87 women, laparoscopy) and tests for the presence of antibody to Chlamydia trachomatis. 73% of the women with distal occlusion of the fallopian tubes and 21% with peritubal adhesions alone had antibodies to C. trachomatis, but none of those with normal tubes did (p less than 0.001 and less than 0.005, respectively). No other infertility factors were associated with an increased frequency of antibodies to C. trachomatis. Since the presence or absence of antibodies to C. trachomatis was as discriminatory in the detection of tubal disease in infertile women as was the hysterosalpingogram, the serological test for C. trachomatis should become part of a routine infertility investigation.

Acute Disease↗
PubMed1982

Effects of corticosteroids and cyclophosphamide on a mouse model of Chlamydia trachomatis pneumonitis.

Suppression of the inflammatory reaction with daily doses of cortisone acetate or cyclophosphamide substantially prolonged the pulmonary infection in mice which had been intranasally inoculated with a trachoma biotype of Chlamydia trachomatis. Titration of organisms recovered from the lungs of treated mice revealed a drop in titer after day 2 (postinfection), followed by a prominent increase on day 6. In cyclophosphamide-treated mice the infection was resolved after day 12, whereas in cortisone acetate-treated mice a significant titer remained after day 17. In contrast, no organisms were recoverable after day 6 in control mice treated with saline or in mice treated with hydrocortisone succinate. Histologically, the ability of cortisone acetate and cyclophosphamide to inhibit the inflammatory reaction correlated with the respective course of chlamydial pneumonitis. This study demonstrated that mice were intrinsically capable of sustaining a lung infection induced by a human strain of S. trachomatis.

Animals↗
PubMed1982

Toxic effect of human polymorphonuclear leukocytes on Chlamydia trachomatis.

The effect of human polymorphonuclear leukocytes (PMNs) on Chlamydia trachomatis was studied. Both trachoma (B/TW-5/OT) and lymphogranuloma venereum (L2/434/Bu) biotypes were rapidly inactivated by exposure to human PMNs. A decrease of 3 to 3.5 logs in viable count was observed after 60 min of incubation at a chlamydia-to-PMN ratio of 1:10. Both chlamydial biotypes were also rapidly inactivated by the cell-free myeloperoxidase-H2O2-halide system. A decrease in infectivity titer of 4 to 5 logs for TW-5 and complete inactivation of 434 were seen after 30 min of incubation. The microbicidal effect was prevented by the deletion of each component of the system or by the addition of the peroxidase inhibitors cyanide or azide. PMNs from myeloperoxidase-deficient patients inactivated chlamydiae normally, whereas PMNs from patients with chronic granulomatous disease, although strongly chlamydicidal, were less effective than normal PMNs in the activation of TW-5 (2-log drop in viable organisms versus a 3 to 3.5-log drop). The chlamydicidal activity of PMNs from patients with chronic granulomatous disease and normal PMNs were comparable against the 434 biotype. These studies suggest that the myeloperoxidase system, or indeed oxygen-dependent antimicrobial systems, are not essential for the chlamydicidal activity of PMNs.

Animals↗
PubMed1982

Sensitivity of immunofluorescence with monoclonal antibodies for detection of Chlamydia trachomatis inclusions in cell culture.

Monoclonal antibodies which recognize the species-specific major outer membrane protein antigen of Chlamydia trachomatis were used for immunofluorescence staining of chlamydial inclusions in cell culture. A total of 115 clinical specimens were inoculated onto replicate HeLa 229 cell monolayers and assayed for chlamydial inclusions by immunofluorescence staining and Giemsa staining. Of the isolates, 38 were detected by immunofluorescence staining on passage 1 and 1 was detected on passage 2; 23 isolates on passage 1 and 13 isolates on passage 2 were detected by Giemsa staining. Immunofluorescence staining was significantly more sensitive than Giemsa staining for detecting chlamydial inclusions, particularly from specimens containing low titers of Chlamydia.

Antibodies, Monoclonal↗
PubMed1982

Therapy of cervical chlamydial infection.

The treatment of cervical Chlamydia trachomatis infection in nonpregnant women was evaluated in a double-blind randomized study. Objective criteria were used to assess the response of cervicitis to therapy. Fifty patients were treated with tetracycline hydrochloride, 500 mg orally four times daily, and 50 patients were treated with rosaramicin, 250 mg orally four times daily, both for 7 days. Both agents were highly effective in eradicating C trachomatis. Both produced significant improvement in objective signs of cervicitis: eliminating mucopurulent endocervical discharge and edema of ectopy, and decreasing the clinical severity score of cervicitis. This trial shows that the 1-week course of tetracycline hydrochloride currently recommended for treatment of chlamydial urethritis in men is also highly effective for the treatment of chlamydial cervical infection in women. Rosaramicin, a macrolide antibiotic, was equally effective but produced a higher rate of gastrointestinal side effects.

Chlamydia Infections↗
PubMed1982

Monoclonal antibodies to Chlamydia trachomatis: antibody specificities and antigen characterization.

Nineteen independent hybrid cell lines that produce monoclonal antibodies to Chlamydia trachomatis surface antigens were prepared by the fusion of mouse myeloma cells with lymphocytes of mice that were immunized with C. trachomatis immunotypes B, C, and L2. Seven serologically distinct reaction patterns were detected by microimmunofluorescence (micro-IF) of elementary body (EB) preparations when culture fluids were tested against a panel of 18 chlamydial serotyping reference strains. These reaction patterns demonstrated genus-, species-, subspecies-, and type-specific distributions. Additionally, these antibodies were tested in parallel against reticulate body (RB) preparations of several chlamydial strains. Monoclonal antibodies that reacted with genus-specific antigens reacted preferentially with RB, whereas antibodies that reacted to species-, subspecies-, or type-specific antigens reacted equivalently to both RB and EB. Physiochemical characterization of antigens recognized by the different monoclonal antibodies was assessed by heat treatment, pronase digestion, periodate oxidation, and immuno-blot techniques. The genus-specific antigen was a heat-stable, pronase-resistant, and relatively periodate-sensitive component of less than 10,000 m.w. The species-, subspecies-, and type-specific antigens were heat stable, pronase sensitive, and periodate resistant. The antibodies that detected species- and subspecies-specific antigens predominantly reacted in immuno-blots with the 40,000 m.w. major outer membrane protein. These monoclonal antibodies now provide a new approach for the precise serologic classification and detection of different C. trachomatis strains.

Animals↗
PubMed1981

Cellular immune response during uncomplicated genital infection with Chlamydia trachomatis in humans.

A lymphocyte transformation (LT) assay for the study of the cellular immune response to Chlamydia trachomatis in humans is described. Subjects studied included 9 newborns whose mothers were C. trachomatis culture negative, 16 seronegative, sexually inexperienced adults, and 107 patients seen at a sexually transmitted disease clinic, including 31 men presenting with acute nongonococcal urethritis and 76 women with known or suspected uncomplicated gonorrhea or with uncomplicated C. trachomatis genital infection. LT stimulation indices (SI) were less than 3.5 in newborns and normal adults, as well as 11 of 12 seronegative, isolation-negative sexually transmitted disease clinic subjects. LTSI greater than 3.5 was found only with subjects who were sero- or culture positive for C. trachomatis. Among men with nongonococcal urethritis, the LTSI correlated better with culture than with antibody. Among women, the LTSI correlated better with antibody than with culture. LTSI decline significantly 3 to 4 weeks after curative therapy in men with nongonococcal urethritis, suggesting that LT response is short-lived and that the LTSI may be an indicator of acute C. trachomatis infection. The sensitivity, specificity, and predictive value of a positive LT assay and of serum and local antibody tests, in terms of C. trachomatis infection defined as positive isolation, were also compared. The predictive value of a positive LTSI for C. trachomatis infection was generally low in the sexually transmitted disease clinic patients: 62% in men with nongonococcal urethritis and 37% in women. However, this study did show the LT assay to be a useful specific test for monitoring the cellular immune response to C. trachomatis infection.

Antibodies, Bacterial↗
PubMed1979

Reticulate bodies as single antigen in Chlamydia trachomatis serology with microimmunofluorescence.

Formalin-fixed, purified reticulate bodies (RB) of Chlamydia trachomatis immunotype C/TW-3/OT were used as a serological test antigen in the microimmunofluorescence test. The sensitivity and specificity of the RB antigen were compared to elementary bodies (EB) used as antigens in the detection of C. trachomatis antibodies in human sera by microimmunofluorescence. RB reacted with all known C. trachomatis immunotypes with the same sensitivity as the homotypic EB. In routine serology with sera and endocervical secretions, the RB antigen had a sensitivity similar to that of the EB in detecting serum antibodies, endocervical secretion antibodies, and antibodies of immunoglobulin M and G classes. No false-positive reactions were detected with control sera. All positive reactions showed type-specific antibodies against an EB immunotype. RB seemed to demonstrate chlamydial group reactivity; sera from 10 psittacosis patients diagnosed clinically and serologically by complement fixation showed five positive, three equivocal, and two negative reactions. By immunofluorescence, RB appeared as distinct rings demonstrating uniform peripheral surface fluorescence at their rims. The EB appeared as pinpoint-sized dots. C/TW-3/OT RB used as a single test anitgen should provide a simple and sensitive serological assay for the detection of C. trachomatis antibody.

Antibodies, Bacterial↗
PubMed1978

Cultures of Chlamydia trachomatis in mouse peritoneal macrophages: factors affecting organism growth.

Growth of Chlamydia trachomatis B/TW-5/OT and L(2)/434/Bu strains in cultures of thioglycolate-activated mouse peritoneal macrophages was studied. Both strains grew to a limited extent in the macrophages, but lymphogranuloma venereum (LGV) grew better than trachoma. Growth was enhanced by centrifugation of the inoculum onto the macrophage cell layer and inhibited by pretreatment of macrophages for 2 h with 100 mug of concanavalin A per ml. No significant effect was observed by pretreatment of macrophages with diethylaminoethyl-dextran (30 mug/ml, 30 min), cytochalasin B (20 mug/ml, 1 h), and cyclophosphamide (200 mug/ml, overnight) or by treatment with hydrocortisone (1 and 10 mug/ml, overnight before inoculation and during a 2-day incubation after inoculation). Resistance to intracellular growth of the two organisms was not increased in macrophages obtained from mice immunized with the organisms compared with macrophages from nonimmunized mice unless they were pretreated in vitro with penicillin (100 U/ml, overnight). The yields of LGV organisms from the penicillin-pretreated macrophages of LGV-immunized mice were 100-fold less than from the pretreated macrophages of nonimmunized control mice. At the same time, the yields of LGV organisms from penicillin-pretreated macrophages of mice immunized with trachoma, gonococcus, and HeLa cells were not different from those obtained in pretreated macrophages of nonimmunized control mice.

Animals↗
PubMed1978

Immediate cytotoxicity of Chlamydia trachomatis for mouse peritoneal macrophages.

The toxicity of Chlamydia trachomatis was studied with mouse peritoneal macrophage culture. Inoculation of 30 inclusion-forming units of trachoma B/TW-5/OT organisms and 250 inclusion-forming units of lymphogranuloma venereum L2/434/Bu organisms per cell caused immediated toxicity, with the killing of 40 to 90% of the macrophages within 6 h after inoculation. Inhibition of phagocytosis by adsorption at 0 degrees C or by NaF pretreatment of macrophages prevented the toxicity, indicating that chlamydiae must be phagocytized to induce toxicity. Infectivity and toxicity could be dissociated, since ultraviolet-inactivated chlamydiae were still toxic. However, the toxicity was destroyed by heating the organisms at 56 degrees C for 10 min. Tetracycline, and antichlamydial drug, did not prevent toxicity, indicating that multiplication of the organisms was not required to induce toxicity. Toxicity was not prevented by treatment of macrophages with hydrocortisone. The toxicity of trachoma TW-5 was reduced by the rabbit immune serum of trachoma TW-5 but not by the rabbit immune serum of psittacosis meningopneumonitis.

Animals↗
PubMed1977

Antimicrobial activity of several antibiotics and a sulfonamide against Chlamydia trachomatis organisms in cell culture.

Minimum inhibitory concentrations of several antibiotics and a sulfonamide for growth of the 15 known immunotypes of Chlamydia trachomatis were determined in HeLa 229 cell cultures. The concentrations for complete inhibition of infectious-organism production were (per milliliter): tetracycline, 0.02 to 0.5 mug; rosamicin, 0.05 to 0.25 mug; erythromycin, 0.1 to 0.5 mug; chloramphenicol, 10 mug; penicillin, 0.02 to 50 U; ampicillin, 0.1 to 50 mug; and sulfisoxazole, 2 to 200 mug. The same concentrations of tetracycline, rosamicin, erythromycin, and chloramphenicol were sufficient to inhibit C. trachomatis inclusion formation. An increased concentration of sulfisoxazole was often needed to inhibit inclusion formation. Penicillin at 100 U/ml and ampicillin at 100 mug/ml failed to completely inhibit inclusion formation.

Anti-Bacterial Agents↗
PubMed1977

Purification of a Chlamydia trachomatis-specific antigen by immunoadsorption with monospecific antibody.

This study describes the isolation and partial characterization of a Chlamydia trachomatis specific antigen. A species-specific antigen of C. trachomatis (antigen-0.65) was identified by two-dimensional immunoelectrophoresis. Antiserum specific for antigen-0.65 was prepared in rabbits by immunizing with agarose-gel precipitates excised from two-dimensional immunoelectrophorograms. Purified gamma-globulins from antigen-0.65 specific serum were coupled to the N-hydroxysuccinimide ester derivative of agarose which was then used for the immunoadsorbent purification of antigen-0.65 from Triton X-100 solubilized lymphogranuloma venereum (L2/434/Bu) organisms. The isolated antigen was immunochemically pure when tested against rabbit antiserum prepared to LGV-434 organisms by using rocket and two-dimensional immunoelectrophoresis. Antigenicity was destroyed by protease treatment and heating at 56 degrees C for 30 min, but the antigen was stable to ribonuclease, deoxyribonuclease, periodate oxidation and pH extremes of 2.2 and 10.6. Polyacrylamide gel electrophoresis of purified antigen showed a major protein band with an apparent m.w. of 155,000.

Animals↗
PubMed1977

Serologic diagnosis of Lymphogranuloma venereum by counterimmunoelectrophoresis with a Chlamydia trachomatis protein antigen.

Previously a protein antigen was identified which was common to members of the species Chlamydia trachomatis and to which antibody could be demonstrated in human sera by two-dimensional immunoelectrophoresis. The antigen has been purified by immunoadsorption chromatography, and an exploration undertaken to determine whether the detection of antibody to this antigen by counterimmunoelectrophoresis would be useful for the diagnosis of C. trachomatis infections. Antibody was found in 43 of 45 (96%) sera from patients with lymphogranuloma venerum proven by either isolation or other serologic methods. No positive reactions were observed in 50 patients with nongonococcal urethritis, 38 of whom had evidence of antibody as measured by microimmunofluorescence with whole organism antigen. Sera were also negative from 50 normal individuals, 10 persons with gonorrhea, and 2 patients with psittacosis. Counterimmunoelectrophoresis with purified C. trachomatis protein antigen appears to be a useful serologic test for diagnosis of lymphogranuloma venereum.

Antigens, Bacterial↗
PubMed1976

Interaction of Chlamydia trachomatis organisms and HeLa 229 cells.

The infection of HeLa 229 cells in monolayer culture with trachoma (B/TW-5/OT) and lymphogranuloma venereum (LGV) (L2/434/Bu) organism was studied in terms of two parameters: radioactivity counts of cell-associated tritium labeled organisms at the initial stage of inoculation for measurement of attachment, and inclusion counts of infection cells after incubation for measurement of growth. Factors affecting attachment and inclusion formation and correlation of the two are presented. It was shown that attachment is an important initial step in infection by Chlamydia trachomatis. The rate of attachment was temperature dependent. The attachment of LGV organisms was affected more profoundly by temperature than was that of trachoma organisms. Attachment and inclusion formation of trachoma and LGV organisms were inhibited by heparin. Diethylaminoethyl-dextran was again shown to enhance attachment and inclusion formation of trachoma but not LGV organisms. NaF had no effect on attachment, but inhibited inclusion formation of both trachoma and LGV organisms. Both attachment and inclusion formation of trachoma organisms were strongly enhanced by centrifugation of the inoculum onto the cell monolayer. Although inclusion formation of trachoma organism was much greater in susceptible cells (HeLa 229) than relatively insusceptible cells (fetal tonsil), attachment was only slightly greater. The results based on the test of two cell lines suggested that attachment prpbably is not a critical factor in determing a cell line's susceptibility to infection with trachoma organisms.

Cell Adhesion↗
PubMed1975

Comparative infectivity of trachoma organisms in HeLa 229 cells and egg cultures.

Thirty-two trachoma strains were simultaneously titrated in the yolk sac of embryonated chicken eggs and in HeLa 229 cells pretreated with diethylaminoethyl-dextran to determine the relative sensitivity of the two culture systems for detection of the infection. The strains tested were both yolk sac- and HeLa cell-established isolates from the eye and the genital tract. The study showed that the cell culture was of equal or greater sensitivity than yolk sac culture and that ocular and genital isolates differ in their ability to propagate in egg and cell culture: genital strains grow better in cell culture than in eggs and ocular strains grow only slightly better in cell culture.

Animals↗
PubMed1975

Comparative susceptibility of eleven mammalian cell lines to infection with trachoma organisms.

Eleven mammalian cell lines, HeLa 229, HeLa M, Hep-2, FT, BHK-21, Vero, MK-2, MPK, L-WO5A2, McCoy, and L-929, were tested for their susceptibility to infection with Trachoma strains TW-3 (type C, ocular origin) and UW-5 (type E, genital origin). All the cell layers were pretreated with diethylaminoethyl-dextran before inoculation of the organisms, and the inocula were centrifuged onto the cell layers. HeLa 229 was found to be the most sensitive to infection as determined by inclusion counts. The next most susceptible were cell lines MK-2, Hep-2, McCoy, and HeLa M, in that order. Infectivity in these cells ranged from 89 to 12% of that observed in HeLa 229. The remaining cell lines, BHK-21, L-WO5A2, L-929, Vero, MPK, and FT, were much less susceptible with infectivity less than 10% that of HeLa 229. HeLa 229 cells and 5-iodo-2'-deoxyuridine-pretreated McCoy cells have been used most extensively with Trachoma organisms in our laboratories. Infectivity in these two cell culture systems, both pretreated with diethylaminoethyl-dextran, was compared using 13 Trachoma strains of both ocular and genital origins of different immunotypes. The two systems performed similarly except with two type C, three type I, and one type J strains, With the type C, I and J strains tested, considerably fewer inclusions were found in 5-iodo-2'-deoxyuridine-pretreated McCoy than in HeLa 229 because inclusion formation of these strains in McCoy cells was not enhanced by 5-iodo-2'-deoxyuridine pretreatment.

Animals↗