scholarly journals Characterization of the Growth of Chlamydia trachomatis in In Vitro-Generated Stratified Epithelium

2017 ◽  
Vol 7 ◽  
Author(s):  
Ana T. Nogueira ◽  
Kristin M. Braun ◽  
Rey A. Carabeo
Keyword(s):  
Chlamydia Trachomatis ◽  
Stratified Epithelium

Characterization of Chlamydia trachomatis antigens with monoclonal and polyclonal antibodies

1988 ◽  
Vol 34 (2) ◽  
pp. 141-147 ◽  
Author(s):  
Ian W. Maclean ◽  
Rosanna W. Peeling ◽  
Robert C. Brunham
Keyword(s):  
Chlamydia Trachomatis ◽  
Polyclonal Antibodies ◽  
Antigenic Specificity ◽  
Immunoblot Assay ◽  
Neutralizing Activity ◽  
Molecular Masses ◽  
Monospecific Antisera ◽  
Biologic Function

We used monoclonal antibodies (MAbs) to examine the antigenic specificity and biologic function of several Chlamydia trachomatis antigens. Thirteen distinct MAbs to eight C. trachomatis antigens were produced. Six MAbs reacted with unique epitopes on the major outer membrane protein (MOMP) and two of these had neutralizing activity. MAbs were produced to each of the chlamydial antigens with molecular masses of 10, 29, 32, 57, 60, 70, and 75 kilodaltons (kDa). These MAbs showed species and genus specificity in an immunoblot assay. None of the MAbs had neutralizing activity. The epitopes recognized on MOMP, 29-, and 10-kDa (presumably lipopolysaccharide) antigens were surface exposed. MAbs to the 75-kDa, 57-kDa, and MOMP antigens were used for immunoaffinity purification of these antigens to produce monospecific antisera in mice. With polyclonal sera, we found that the 75-kDa antigen was also immunoaccessible and that antibody to MOMP and 75-kDa antigens neutralized C. trachomatis infectivity. We conclude that, in addition to MOMP and lipopolysaccharide, antigens with molecular masses of 75 and 29 kDa are surface exposed. Antibodies to MOMP and 75-kDa antigens can neutralize the organism in vitro.

scholarly journals Characterization of In Vitro Chlamydial Cultures in Low-Oxygen Atmospheres

2007 ◽  
Vol 189 (18) ◽  
pp. 6723-6726 ◽  
Author(s):  
Nicolai Juul ◽  
Helene Jensen ◽  
Malene Hvid ◽  
Gunna Christiansen ◽  
Svend Birkelund
Keyword(s):  
Chlamydia Trachomatis ◽  
Chlamydia Pneumoniae ◽  
Low Oxygen ◽  
Atmospheric Air ◽  
Chlamydial Infections

ABSTRACT To mimic in vivo conditions during chlamydial infections, Chlamydia trachomatis serovar D and Chlamydia pneumoniae CWL029 were cultured in low-oxygen atmospheres containing 4% O2, with parallel controls cultured in atmospheric air. Both were enriched with 5% CO2. The results showed a dramatic increase in the growth of C. pneumoniae but not of C. trachomatis.

scholarly journals The Chlamydia trachomatis Plasmid Is a Transcriptional Regulator of Chromosomal Genes and a Virulence Factor

2008 ◽  
Vol 76 (6) ◽  
pp. 2273-2283 ◽  
Author(s):  
John H. Carlson ◽  
William M. Whitmire ◽  
Deborah D. Crane ◽  
Luke Wicke ◽  
Kimmo Virtaneva ◽  
...  
Keyword(s):  
Chlamydia Trachomatis ◽  
Glycogen Synthase ◽  
Human Infection ◽  
Primary Role ◽  
Phenotypic Differences ◽  
Chromosomal Genes ◽  
Movement Characteristic

ABSTRACT Chlamydia trachomatis possesses a cryptic 7.5-kb plasmid of unknown function. Here, we describe a comprehensive molecular and biological characterization of the naturally occurring plasmidless human C. trachomatis strain L2(25667R). We found that despite minimal chromosomal polymorphisms, the LGV strain L2(25667R) was indistinguishable from plasmid-positive strain L2(434) with regard to its in vitro infectivity characteristics such as growth kinetics, plaquing efficiency, and plaque size. The only in vitro phenotypic differences between L2(434) and L2(25667R) were the accumulation of glycogen granules in the inclusion matrix and the lack of the typical intrainclusion Brownian-like movement characteristic of C. trachomatis strains. Conversely, we observed a marked difference between the two strains in their abilities to colonize and infect the female mouse genital tract. The 50% infective dose of plasmidless strain L2(25667R) was 400-fold greater (4 × 106 inclusion-forming units [IFU]) than that of plasmid-bearing strain L2(434) (1 × 104 IFU). Transcriptome analysis of the two strains demonstrated a decrease in the transcript levels of a subset of chromosomal genes for strain L2(25667R). Among those genes was glgA, encoding glycogen synthase, a finding consistent with the failure of L2(25667R) to accumulate glycogen granules. These findings support a primary role for the plasmid in in vivo infectivity and suggest that virulence is controlled, at least in part, by the plasmid's ability to regulate the expression of chromosomal genes. Our findings have important implications in understanding a role for the plasmid in the pathogenesis of human infection and disease.

scholarly journals Genomic and phenotypic characterization of in vitro-generated Chlamydia trachomatis recombinants

2013 ◽  
Vol 13 (1) ◽  
pp. 142 ◽  
Author(s):  
Brendan M Jeffrey ◽  
Robert J Suchland ◽  
Steven G Eriksen ◽  
Kelsi M Sandoz ◽  
Daniel D Rockey
Keyword(s):  
Chlamydia Trachomatis ◽  
Phenotypic Characterization

Assays for Phospholipid-Dependent Formation of Thrombin and Xa: A Potential Method for Quantifying Lupus Anticoagulant Activity

1991 ◽  
Vol 66 (04) ◽  
pp. 453-458 ◽  
Author(s):  
John T Brandt
Keyword(s):  
Specific Activity ◽  
Anticoagulant Activity ◽  
Factor Xa ◽  
Clinical Importance ◽  
Potential Method ◽  
Quantitative Expression ◽  
Increased Risk ◽  
Apparent Association

SummaryLupus anticoagulants (LAs) are antibodies which interfere with phospholipid-dependent procoagulant reactions. Their clinical importance is due to their apparent association with an increased risk of thrombo-embolic disease. To date there have been few assays for quantifying the specific activity of these antibodies in vitro and this has hampered attempts to purify and characterize these antibodies. Methods for determining phospholipid-dependent generation of thrombin and factor Xa are described. Isolated IgG fractions from 7 of 9 patients with LAs were found to reproducibly inhibit enzyme generation in these assay systems, permitting quantitative expression of inhibitor activity. Different patterns of inhibitory activity, based on the relative inhibition of thrombin and factor Xa generation, were found, further substantiating the known heterogeneity of these antibodies. These systems may prove helpful in further purification and characterization of LAs.

Characterization of the Original Christmas Disease Mutation (Cysteine 206 → Serine): From Clinical Recognition to Molecular Pathogenesis

1992 ◽  
Vol 67 (01) ◽  
pp. 063-065 ◽  
Author(s):  
Sherryl A M Taylor ◽  
Jacalyn Duffin ◽  
Cherie Cameron ◽  
Jerome Teitel ◽  
Bernadette Garvey ◽  
...  
Keyword(s):  
Nucleotide Substitution ◽  
Novel Mutation ◽  
Factor Ix ◽  
Causative Mutation ◽  
Coding Region ◽  
Body Of Knowledge ◽  
Polymerase Chain ◽  
Splice Junctions

SummaryChristmas disease was first reported as a distinct clinical entity in two manuscripts published in 1952 (1, 2). The eponym associated with this disorder, is the surname of the first patient examined in detail and reported by Biggs and colleagues in a paper describing the clinical and laboratory features of seven affected individuals (3). This patient has severe factor IX coagulant deficiency (less than 0.01 units/ml) and no detectable circulating factor IX antigen (less than 0.01 units/ml). Coding sequence and splice junctions of the factor IX gene from this patient have been amplified in vitro through the polymerase chain reaction (PCR). One nucleotide substitution was identified at nucleotide 30,070 where a guanine was replaced by a cytosine. This mutation alters the amino acid encoded at position 206 in the factor IX protein from cysteine to serine. The non conservative nature of this substitution, the absence of this change in more than 200 previously sequenced factor IX genes and the fact that the remainder of the coding region of this gene was normal, all provide strong circumstantial evidence in favour of this change being the causative mutation in this patient. The molecular characterization of this novel mutation in the index case of Christmas disease, contributes to the rapidly expanding body of knowledge pertaining to Christmas disease pathogenesis.

IN VITRO/IN SILICO CHARACTERIZATION OF ACTIVE AND PASSIVE STRESSES IN CARDIAC MUSCLE

2012 ◽  
Vol 10 (2) ◽  
pp. 171-188 ◽  
Author(s):  
Markus Boel ◽  
Oscar J. Abilez ◽  
Ahmed N Assar ◽  
Christopher K. Zarins ◽  
Ellen Kuhl
Keyword(s):  
Cardiac Muscle ◽  
In Silico ◽  
In Silico Characterization
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