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C C Kuo

Publications and source records attributed to C C Kuo.

At least 199 records · Page 11Linked to original sources

Immunotyping of Chlamydia trachomatis with monoclonal antibodies.

Rapid and precise immunotyping of Chlamydia trachomatis was achieved by testing whole organisms (elementary bodies) in the microimmunofluorescence test with monoclonal antibodies. Monoclonal antibodies were produced with standard techniques by using an immunization schedule that encouraged the development of immunotype-specific antibodies. Fifteen monotypic or multitypic (subspecies) monoclonal antibodies were chosen for use in a two-step typing system that required strains of C. trachomatis to be tested against six to eight monoclonal antibodies for classification. Immunotyping with monoclonal antibodies was studied by testing 313 strains, typed with the previous method that utilized immunized mouse antisera, that represented each of the 15 established serovars. The two-step monoclonal antibody method resulted in a classification similar to the current one. Only one strain typed differently with the two methods. With the monoclonal antibody method, available lymphogranuloma venereum (LGV) serovars L1 and L3 could not be differentiated from trachoma serovars E and G, respectively, unless the strains had been identified as LGV. Monoclonal antibody typing was simpler to perform and more precise; it allowed easy differentiation between closely related serovars. Three new types were discovered among the strains previously classified as serovars D, I, and L2. These are tentatively being considered subtypes and are labeled D', I', and L2'.

Animals↗

Detection of multiple serovars of Chlamydia trachomatis in genital infections.

Eight genital isolates of Chlamydia trachomatis that each contained two different serovars were identified by typing with serovar-specific monoclonal antibodies. Mixed infections were confirmed by using serovar-specific monoclonal antibodies in an indirect immunofluorescence assay. For some isolates, fluorescein/rhodamine double-label indirect immunofluorescence with mixtures of IgM and IgG monoclonal antibodies to Chlamydia provided direct visual confirmation of mixed infection. Two isolates contained serovars D and F, and two E and F; one isolate contained serovars Ba and E, one D and J, one I' and F, and one E and J. In a sample of 352 genital isolates of C. trachomatis consecutively typed by dot ELISA with monoclonal antibodies, seven (2%) demonstrated mixed serovars.

Antibodies, Bacterial↗

Molecular cloning and expression of Chlamydia trachomatis major outer membrane protein antigens in Escherichia coli.

DNA obtained from Chlamydia trachomatis (serovar L2) was partially digested with DNase I and inserted into the beta-galactosidase gene of bacteriophage lambda gt11. Seven recombinants were selected that produced immunoreactive fusion proteins which were detected with anti-C. trachomatis rabbit serum. One recombinant, designated lambda gt11/L2/33, reacted with various monoclonal antibodies that recognize species-, subspecies-, and type-specific determinants on the chlamydial major outer membrane protein (MOMP). Immunoblot analysis of a lambda gt11/L2/33 lysogen revealed a fusion protein that expressed a approximately 15,000-dalton carboxyl-terminal peptide of the chlamydial MOMP. This moiety of the MOMP possesses epitopes responsible for each of the unique reactivities demonstrated by anti-MOMP monoclonal antibodies. The lambda gt11/L2/33 recombinant contained a 1.1-kilobase DNA insert which hybridized to DNA isolated from each of the 15 C. trachomatis serovars.

Antibodies, Monoclonal↗

Neutralization of Chlamydia trachomatis cell culture infection by serovar-specific monoclonal antibodies.

Seventeen monoclonal antibodies (MAs) were tested. Five of seven serovar-specific MAs from five different serovars and two of five subspecies-specific MAs showed neutralizing activity as well as serovar specificity. No species- or genus-specific MAs showed neutralizing activity. No neutralization occurred without complement. Results indicate neutralization was serovar specific and complement dependent.

Animals↗

Rapid immunotyping of Chlamydia trachomatis with monoclonal antibodies in a solid-phase enzyme immunoassay.

The technical complexity of determining the serovar of Chlamydia trachomatis strains has limited the use of serotyping in clinical and epidemiologic studies. We developed a simple method for rapidly serotyping isolates of C. trachomatis by using monoclonal antibodies in a dot-enzyme-linked immunosorbent assay (ELISA) system. Isolates were passaged three to six times in shell vial cultures to greater than 50% monolayer infection, and chlamydial elementary bodies were isolated by sonication and microcentrifugation. Chlamydial antigen was spotted onto a series of replicate nitrocellulose membrane patches and reacted with C. trachomatis-specific monoclonal antibodies. Bound antibody was detected visually by a color reaction by using peroxidase-conjugated anti-mouse immunoglobulins. This method can be routinely applied to 60 or more specimens concurrently. We compared dot-ELISA serotyping with monoclonal antibody microimmunofluorescence serotyping of 124 clinical C. trachomatis isolates and found that dot-ELISA has sensitivity and serotyping accuracy comparable to that of monoclonal antibody microimmunofluorescence.

Antibodies, Bacterial↗

Mucopurulent cervicitis--the ignored counterpart in women of urethritis in men.

Among 100 randomly selected nonmenstruating women attending a clinic for sexually transmitted diseases, we assessed objective criteria for the clinical diagnosis of mucopurulent cervicitis. Visualization of yellow mucopurulent endocervical secretions on a white swab and the presence of 10 or more polymorphonuclear leukocytes per microscopical field (at a magnification of 1000) in satisfactory gram-stained endocervical smears were independently correlated with cervical Chlamydia trachomatis infection. Neither finding correlated with gonorrhea or genital herpes, although herpes caused characteristic cervical ulcerations. C. trachomatis was isolated from the cervix of 20 of 40 women with mucopurulent cervicitis but of only 2 of 60 without it. The overall prevalence of mucopurulent cervicitis among women attending the clinic (40 per cent) exceeded that of nongonococcal urethritis among men in the same clinic, and the prevalence of C. trachomatis infection was higher in mucopurulent cervicitis than in nongonococcal urethritis, a condition that is conventionally treated with tetracyclines. These findings support recommendations for the treatment of mucopurulent cervicitis and should guide the selective use of confirmatory diagnostic tests for C. trachomatis infection.

Adolescent↗

Culture-independent diagnosis of Chlamydia trachomatis using monoclonal antibodies.

To simplify the diagnosis of chlamydial genital infection, we used a fluorescein-conjugated monoclonal antibody in immunofluorescence tests on smears prepared from urethral or cervical secretions obtained directly from patients. This direct test, requiring less than 30 minutes to perform, was based on the detection of extracellular chlamydial elementary bodies. A comparison of the direct test with cultures stained with iodine on specimens from 926 patients demonstrated a sensitivity of 93 per cent and a specificity of 96 per cent. The direct test provides a rapid, simple, and sensitive method for the diagnosis of chlamydial infection, which can be performed in laboratories that do not have tissue-culture capability.

Antibodies, Monoclonal↗

Microbial causes of neonatal conjunctivitis.

We assessed the microbial causes of neonatal conjunctivitis by comparing 55 infants with purulent conjunctivitis and 60 healthy control infants. A mean of greater than 5 leukocytes per 1000X microscopic field was seen in Gram-stained smears obtained from the more inflamed eye in 77% of 30 untreated patients but none of 57 controls. Pathogens isolated more often from untreated patients than from controls included Haemophilus spp. (17% vs 2%, P = 0.01), Staphylococcus aureus (17% vs 2%, P = 0.01), Chlamydia trachomatis (14% vs 0%, P = 0.01), enterococci (8% vs 0%, P = 0.05), and Streptococcus pneumoniae (11% vs 2%, P = 0.06). One or more of these pathogens were isolated from the conjunctivae in 58% of patients and 5% of controls (P less than 0.001). Bacterial morphology seen on smear correlated with the pathogens cultured. Isolation of Haemophilus spp. or S. pneumoniae was associated with dacryostenosis. We conclude that several microbial pathogens are implicated in neonatal conjunctivitis. These organisms have differing susceptibilities to antimicrobial agents, so culture and sensitivity testing are required as a guide to therapy.

Bacterial Infections↗

Rapid diagnosis of Chlamydia trachomatis pneumonia in infants.

By utilizing a species specific monoclonal antibody against Chlamydia trachomatis for a direct fluorescent antibody stain, chlamydia organisms were demonstrated in smears of respiratory tract secretions from two infants with pneumonia. C. trachomatis organisms were isolated from tracheo-nasopharyngeal aspirates of both babies and high titer IgM antibodies found in both against the isolated serotype. None of the three diagnostic methods were positive for C. trachomatis in any of the other 39 infants and children to six years of age with pneumonia studied.

Antibodies, Monoclonal↗

Chlamydia trachomatis species-specific epitope detected on mouse biovar outer membrane protein.

A common antigenic determinant on the chlamydial major outer membrane protein was detected on each of the three Chlamydia trachomatis biovars (trachoma, lymphogranuloma venereum, and mouse). This determinant was prominently displayed on the surface of chlamydial strains from both the trachoma and lymphogranuloma venereum biovars. However, detection of this determinant on a mouse biovar strain required denaturation by sodium dodecyl sulfate or periodate oxidation. This determinant provides a definable taxonomic link between the three biovars of C. trachomatis.

Animals↗

Two-dimensional echocardiographic detection of pericardiocentesis-induced intrapericardial thrombus.

Heart puncture-induced hemopericardium is a serious complication of the pericardiocentesis. Two-dimensional echocardiographic examination performed in two patients just before and after the development of hemopericardium revealed a unique image in which pericardial bleeding manifested itself by an echodensity thrombus appearance adjacent to the cardiac chambers. With appropriate differentiation to some other intrapericardial echodensity masses, such distinctive images can be highly specific for active bleeding into the pericardial sac.

Aged↗

Chlamydia trachomatis infections in men with Reiter's syndrome.

Chlamydia trachomatis was isolated from 9 of 19 men with acute, nondiarrheal Reiter's syndrome who had not recently taken antibiotics. None of 8 untreated men with other forms of arthritis were infected with this organism. Chlamydia trachomatis-specific antibody titers and cellular immune responses were positive significantly more often in 35 treated and untreated men with acute, nondiarrheal Reiter's syndrome than in 7 men with diarrhea-associated Reiter's syndrome and 8 men with other forms of arthritis. Mean peak chlamydial antibody titers and mean lymphocyte transformation stimulation indices were significantly higher in C. trachomatis-infected men with Reiter's syndrome than in C. trachomatis-infected men with uncomplicated nongonococcal urethritis. We concluded that C. trachomatis is capable of "triggering" Reiter's syndrome in susceptible men and that an exaggerated immune response to this organism may play a role in the pathogenesis of the disease.

Acute Disease↗

Direct fluorescent monoclonal antibody stain for rapid detection of infant Chlamydia trachomatis infections.

A method of direct fluorescent antibody staining for rapid diagnosis of Chlamydia trachomatis infections in infants is described. This method utilized a fluorescein-conjugated species-specific monoclonal antibody to C trachomatis for detecting chlamydial elementary bodies in smears of the conjunctiva, nasopharynx, oropharynx, anus, and vagina. The sensitivity of direct fluorescent antibody staining was compared with isolation of the organisms in McCoy cells. Thirty-nine infants with purulent conjunctivitis were studied. Diagnosis of C trachomatis conjunctivitis was correctly made by smear in all 16 infants when inflamed eyes were sampled. Positive smears were obtained from 12/14 culture-positive and 4/16 culture-negative nasopharyngeal specimens from infants with chlamydial conjunctivitis. All nasopharyngeal cultures and smears from infants with nonchlamydial conjunctivitis were negative. These results indicate that the direct smear test is a sensitive and specific test for diagnosing C trachomatis infection of the eye and nasopharynx in infants, and this test can be completed within one hour of specimen collection.

Antibodies, Monoclonal↗

Reactivation of Chlamydia trachomatis lung infection in mice by cortisone.

To study the latency, chronicity, and recurrent nature of chlamydial infection, we attempted to reactivate Chlamydia trachomatis lung infection in mice by immunosuppressive therapy with cortisone. Mice were treated with subcutaneous injections of cortisone acetate (125 mg/kg) every other day, starting on day 14 after intranasal inoculation of C. trachomatis serotype B (TW-5). C. trachomatis was recovered from the lungs beginning day 6 after the start of cortisone treatment until the end of the observation period on day 12 of treatment. Overall, the reactivation was successful in 8 of 55 mice treated with cortisone, in contrast to 0 of 41 inoculated, untreated mice (P = 0.009) and 0 of 35 uninoculated, treated mice. Cortisone treatment affected the ability of peritoneal exudate cells to respond to migratory inhibition after exposure to purified whole organisms of C. trachomatis serotype B (TW-5) but had little effect on serum antibody titers, indicating a possible role for cellular immunity in resistance against C. trachomatis infection in the lung.

Animals↗

Correlation of host immune response with quantitative recovery of Chlamydia trachomatis from the human endocervix.

We studied 95 women with uncomplicated Chlamydia trachomatis cervical infection. Quantitative isolation of C. trachomatis was performed in HeLa 229 cells, and the results were correlated with serum immunoglobulin M and immunoglobulin G antibody to the organism. We found that quantitative cultures for C. trachomatis can provide a meaningful measurement by which to evaluate the effect of the acquired immune response. In particular, secretory immunoglobulin A antibody to C. trachomatis in cervical secretion demonstrated a striking and inverse correlation with recovery of the organism from the cervix. It is suggested that this component of the immune response may regulate shedding of the organism.

Antibodies, Bacterial↗

Immunotypes of Chlamydia trachomatis isolates in Seattle, Washington.

The immunotype distribution of 493 Chlamydia trachomatis isolates from 467 adults and 26 infants in the Seattle area from 1965 to 1982 is presented. All except one of the isolates from adults were from the genitourinary tract, rectum, or bubo. The proportions of each immunotype were ED, 46.5%; GF, 24.6%; H, I, J/CJ, and K, 5 to 7% each; B, 3.5%; and L2, 1%.

Adult↗

Depression of the lymphocyte transformation response to microbial antigens and to phytohemagglutinin during pregnancy.

Lymphocyte transformation (LT) responses to Chlamydia trachomatis, to four other microbial antigens, and to phytohemagglutinin (PHA) were studied in 201 women during pregnancy and/or 3-18 wk postpartum. The LT responses to all stimulants tested were significantly depressed during pregnancy when compared with postpartum LT responses. This difference occurred whether LT assays were performed in autologous or pooled heterologous plasma collected from nonpregnant donors. Among women studied in the third trimester and again postpartum, the autologous LT stimulation index (LTSI) rose from 1.7 to 3.4 (P less than 0.001) with C. trachomatis elementary body antigen, from 3.7 to 7.9 (P less than 0.001) with Candida albicans cell wall extract, from 4.5 to 7.8 (P = 0.008) with streptokinase-streptodornase, from 1.7 to 3.0 (P = 0.007) with fluid tetanus toxoid, from 1.7 to 2.8 (P = 0.046) with mumps virus skin test antigen, from 35.5 to 87.0 (P less than 0.001) with PHA (2 micrograms/ml), and from 107.2 to 181.9 (P = 0.007) with PHA (10 micrograms/ml). LT responses to C. trachomatis were compared in 52 pregnant women and 58 nonpregnant women; all the women had C. trachomatis isolated at the time of LT assay. Using either plasma supplement, the mean LTSI with C. trachomatis antigen was significantly higher in nonpregnant women than in pregnant women, regardless of trimester (P less than 0.001). Among 12 women who were serially tested and remained culture positive for C. trachomatis throughout pregnancy and the postpartum period, the mean autologous LTSI rose from 1.9 in the third trimester to 7.8 postpartum (P = 0.0004). These data are the first to show that the immune response to an ongoing bacterial infection is depressed during pregnancy and to definitively document the depressed LT responses during human pregnancy.

Antigens, Bacterial↗