Restriction endonuclease analysis of DNA from Chlamydia trachomatis biovars

Peterson, Ellena M.; Maza, Luis M. de la

doi:10.1128/jcm.26.4.625-629.1988

Cited by 34 publications

(19 citation statements)

References 16 publications

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“…In Table 2 are the neutralization data from the high-titer patient no. 4 convalescent-phase sera against various serotypes and strains of LGV L2 UCI-20 6 9 ± 17 9 ± 9 0 ± 12 L2 ATCC-434 4 a Both the isolate and serum were collected on the same day from the patient. All assays were performed twice on different days, with the exception of those for strain MI-2225.…”

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confidence: 99%

“…In animal models they are more lethal and in cell cultures they are easier to propagate than the trachoma biovar strains, probably due to their invasive properties (2,8). These two biovars also differ in their DNA structure as shown by restriction endonuclease analysis (3,4) and their outer membrane proteins with regard to charge (60 and 12 kilodaltons) and molecular mass (12 kilodaltons) (1).…”

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Differences in susceptibilities of the lymphogranuloma venereum and trachoma biovars of Chlamydia trachomatis to neutralization by immune sera

et al. 1990

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Sera from seven patients from whom a C. trachomatis serovar L2 strain was isolated were tested in vitro for their ability to neutralize the infectivity of this organism. In one patient an inguinal lymph node was culture positive, whereas the remaining six patients had positive rectal biopsies. Sera from four of the patients, including the patient with the lymph node isolate, failed to neutralize serovar L2(434). In addition, the homologous strain recovered from the inguinal lymph node was available and was resistant to neutralization by the homologous sera. However, the same sera effectively neutralized a trachoma serovar, E(Bour). All four sera had inclusion immunofluorescent-antibody titers to C. trachomatis serovar L2 of 2,048 to 16,384 and microimmunofluorescent-antibody titers to the lymphogranuloma venereum biovar were equal or higher in all cases than to the 12 serovars of the trachoma biovar. The three remaining sera, while neutralizing the infectivity of the L2 strains tested, neutralized serovar E to a greater extent. These sera had the same inclusion immunofluorescent antibody titers as the sera that failed to neutralize serovar L2. To see whether this difference in the sensitivity of the biovars toward neutralization could be characterized, sera were obtained from mice immunized with different doses of both serovars L2 and E. Sera obtained from mice immunized with serovar E were able to effectively neutralize the homologous strain. In contrast, neutralization of the immunizing strain, L2(UCI-20), was not seen with sera obtained on days 7, 14, and 21 after immunization from animals receiving 8 x 105 and 8 x 104 inclusion-forming units of L2(UCI-20); however, these same sera neutralized serovar E. However, with a higher immunizing dose of L2 (107 IFUs), both E and L2 were neutralized with sera obtained 7 and 14 days after immunization. Therefore, the relative resistance to neutralization by serovar L2 compared with that of serovar E in the mouse model was inoculum dependent. All stock strains were raised in either HeLa 229 cells or McCoy cells and frozen in a sucrose-phosphate glutamic acid buffer (2-SPG) at -70°C (6). Clinical isolates were collected, placed in 2-SPG, and cultured on McCoy or HeLa 229 cells. To obtain high titers, strains were passed at least four times on HeLa 229 or McCoy cells before being frozen in 2-SPG at -70°C. Clinical isolates were serotyped by microimmunofluorescence (MIF) with monoclonal antibodies (11).Chlamydia serology. Sera obtained from patients were assayed by two different indirect immunofluorescent assays. The MIF assay used elementary bodies of the 15 serovars of C. trachomatis as the antigens as previously described (5). In the other method, which will be referred to as the inclusion indirect immunofluorescence assay (IFA), acetone-fixed McCoy cells infected with serovar L2(434) were used (5). Dilutions of sera in phosphate-buffered saline (0.01 M, pH 7.4) were placed on the antigen-containing slides and allowed to incubate at 37°C for 30 min, followed by washing...

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Section: Resultsmentioning

confidence: 99%

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confidence: 99%

Differences in susceptibilities of the lymphogranuloma venereum and trachoma biovars of Chlamydia trachomatis to neutralization by immune sera

et al. 1990

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“…C. trachomatis serovars Li (440), L2 (434), L3 (404), A (G-17), B (TW-5), Ba (Apache-2), C (TW-3), D (IC-Cal-8), E (Bour), F (UW-6), G (UW-57), H (UW-4), I (UW-12), J (UW-36), and K (UW-31); Chlamydia psittaci (Texas turkey); and TWAR-183 were grown in HeLa 229 cells (American Type Culture Collection, Rockville, Md.) as previously described (17)(18)(19)22).…”

Section: Methodsmentioning

confidence: 99%

“…C. trachomatis L3 EBs were purified as described by Caldwell et al (3). The DNA was extracted from purified EBs by using the method of Peterson and de la Maza (17,18), and a library was constructed in Agtll (27). The L3 DNA was sonicated for 10 s (Braun Sonic, 2,000; B. Braun Instruments, Burlingame, Calif.), blunt ended with Escherichia coli polymerase I (New England BioLabs, Inc., Beverly, Mass.…”

Section: Methodsmentioning

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Cloning and characterization of a Chlamydia trachomatis L3 DNA fragment that codes for an antigenic region of the major outer membrane protein and specifically hybridizes to the C- and C-related-complex serovars

1989

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Chlamydia trachomatis L3 DNA was cloned and expressed in Agtll. A recombinant plaque that expressed an antigen that reacted with rabbit polyclonal antichlamydial L3 serum and with two monoclonal antibodies specific for serovars L3 and I was selected from this Chiamydia genomic library. The I-galactosidase Chlamydia fusion protein was purified by immunoaffinity chromatography and injected into mice to produce monoclonal antibodies. These monoclonal antibodies reacted by Western (immuno-) blot with both the fusion protein and the major outer membrane protein from purified L3 elementary bodies. The chlamydial DNA fragment was shown by DNA sequence analysis to be 168 base pairs in length and to correspond to the constant regions 1 and 2 and the variable segment 1 of the major outer membrane protein gene. The recombinant chlamydial DNA fragment hybridized under stringent conditions by Southern and dot blot analysis exclusively with the DNA from the Cand C-related-complex C. trachomatis serovars.Production of L3 rabbit hyperimmune antisera and mouse monoclonal antibodies to C. trachomatis elementary bodies (EBs). L3 antiserum was produced in a female New Zealand White rabbit (Simmonsen Laboratories, Inc., Gilroy, Calif.) by using Renografin 76 (E. R. Squibb and Sons, Inc., Princeton, N.J.)-purified L3 EBs and the injection schedule outlined by Caldwell and Schachter (4). Monoclonal antibodies 487 on August 3, 2020 by guest http://iai.asm.org/ Downloaded from LITERATURE CITED 1. Allan, I., T. M. Cunningham, and M. A. Lovett. 1984. Molecular cloning of the major outer membrane protein of Chlamydia trachomatis. Infect. Immun. 45:637-641. 2. Baehr

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Effects of interferon gamma onChlamydia trachomatis serovar A and L2 protein expression investigated by two-dimensional gel electrophoresis

1999

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Restriction endonuclease analysis of DNA from Chlamydia trachomatis biovars

Cited by 34 publications

References 16 publications

Differences in susceptibilities of the lymphogranuloma venereum and trachoma biovars of Chlamydia trachomatis to neutralization by immune sera

Differences in susceptibilities of the lymphogranuloma venereum and trachoma biovars of Chlamydia trachomatis to neutralization by immune sera

Cloning and characterization of a Chlamydia trachomatis L3 DNA fragment that codes for an antigenic region of the major outer membrane protein and specifically hybridizes to the C- and C-related-complex serovars

Effects of interferon gamma onChlamydia trachomatis serovar A and L2 protein expression investigated by two-dimensional gel electrophoresis

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