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Infection of Human Retinal Pigment Epithelium with Chlamydia trachomatis.

Boiko E, Maltsev D, Savicheva A, Shalepo K, Khusnutdinova T, Pozniak A, Kvetnoi I, Polyakova V, Suetov A - PLoS ONE (2015)

Bottom Line: All eight clinical isolates demonstrated ability to infect hRPE cells.At 24h PI, the percentage of inclusion-containing cells varied from 1.5 ± 0.52 to 14.6 ± 3.3% in hRPE cell culture infected at MOI of 2.0 against 0.37 ± 0.34 to 8.9 ± 0.2% in McCoy cell culture infected at MOI of 0.3.Collagen type I, collagen type IV, basic fibroblast growth factor, transforming growth factor-beta and interleukin-8 expression at 48h PI were maximally increased, by 2.1-, 1.3-, 1.5-, 1.5- and 1.6-fold, respectively, in the Chlamydia trachomatis-infected compared with control hRPE cell culture specimens (P < 0.05).

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Military Medical Academy, St Petersburg, Russia.

ABSTRACT

Purpose: Little is known about the susceptibility of posterior segment tissues, particularly the human retinal pigment epithelium (hRPE), to Chlamydia trachomatis. The purpose of the study was to investigate the possibility of infecting the hRPE with Chlamydia trachomatis, and to examine the infectivity of different Chlamydia trachomatis clinical isolates for hRPE cells and the hRPE cell response to the infection.

Methods: Cultured hRPE and McCoy cells were inoculated with eight Chlamydia trachomatis (serovar E) clinical isolates at multiplicity of infection (MOI) of 2.0 or 0.3. To detect Chlamydia trachomatis, samples were stained immunohistochemically with anti-major outer membrane protein antibodies at 24h, 48h, and 72h postinoculation (PI). The changes in the expression of signaling molecules and proteins of cytoskeleton and extracellular matrix in hRPE cells were examined immunohistochemically.

Results: All eight clinical isolates demonstrated ability to infect hRPE cells. At equal MOI of 0.3, the infectivity of Chlamydia trachomatis clinical isolates for RPE culture was found to be at least as high as that for McCoy cell culture. At 24h PI, the percentage of inclusion-containing cells varied from 1.5 ± 0.52 to 14.6 ± 3.3% in hRPE cell culture infected at MOI of 2.0 against 0.37 ± 0.34 to 8.9 ± 0.2% in McCoy cell culture infected at MOI of 0.3. Collagen type I, collagen type IV, basic fibroblast growth factor, transforming growth factor-beta and interleukin-8 expression at 48h PI were maximally increased, by 2.1-, 1.3-, 1.5-, 1.5- and 1.6-fold, respectively, in the Chlamydia trachomatis-infected compared with control hRPE cell culture specimens (P < 0.05).

Conclusions: This study, for the first time, proved the possibility of infecting hRPE cultured cells with Chlamydia trachomatis, which leads to proproliferative and proinflammatory changes in the expression of signaling molecules and extracellular matrix components.

No MeSH data available.


Related in: MedlinePlus

Immunohistochemistry and direct immunofluorescent staining of hRPE and McCoy cell cultures at different time-points postinoculation with Chlamydia trachomatis (clinical isolate No.24032).In both types of cultures, both techniques reveal highly immunoreactive inclusions (arrowheads). Twenty-four to 72 h postinoculation, a reduction in the number of intracellular inclusions is observed due to the release of a new generation of EBs to the extracellular environment. Scale bar: 50 μm.
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pone.0141754.g001: Immunohistochemistry and direct immunofluorescent staining of hRPE and McCoy cell cultures at different time-points postinoculation with Chlamydia trachomatis (clinical isolate No.24032).In both types of cultures, both techniques reveal highly immunoreactive inclusions (arrowheads). Twenty-four to 72 h postinoculation, a reduction in the number of intracellular inclusions is observed due to the release of a new generation of EBs to the extracellular environment. Scale bar: 50 μm.

Mentions: In all non-control samples of RPE and McCoy cell cultures, the pathogen was detected both by immunohistochemistry and by direct immunofluorescent staining (Fig 1). Twenty-four hours postinoculation (hpi), clinical isolates (at equal multiplicity of infection (MOI) of 2.0) demonstrated a wide variation in infectivity, with the percentage of RPE culture inclusion-containing cells ranging from 1.5±0.52% (isolate No.3438) to 14.6±3.3% (isolate No.24032), whereas that of McCoy culture cells infected at equal MOI of 2.0 was 0.37±0.34% (isolate No.1609) to 8.9±2.4% (isolate No.24032) (Fig 2). Twenty-four hpi, the variation in the percentage of RPE culture inclusion-containing cells among five clinical isolates used for infection at MOI 2.0 was almost 10-fold (1.5–14.6%). All the five clinical isolates used at a higher MOI for infection of RPE cells than for infection of McCoy cells showed a significantly higher (P<0.05) percentage of inclusion-containing cells for hRPE cell culture than for McCoy cell culture. At equal MOI of 0.3, no statistically significant difference in the percentage of inclusion-containing cells was found between McCoy cells and RPR cells.


Infection of Human Retinal Pigment Epithelium with Chlamydia trachomatis.

Boiko E, Maltsev D, Savicheva A, Shalepo K, Khusnutdinova T, Pozniak A, Kvetnoi I, Polyakova V, Suetov A - PLoS ONE (2015)

Immunohistochemistry and direct immunofluorescent staining of hRPE and McCoy cell cultures at different time-points postinoculation with Chlamydia trachomatis (clinical isolate No.24032).In both types of cultures, both techniques reveal highly immunoreactive inclusions (arrowheads). Twenty-four to 72 h postinoculation, a reduction in the number of intracellular inclusions is observed due to the release of a new generation of EBs to the extracellular environment. Scale bar: 50 μm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4633144&req=5

pone.0141754.g001: Immunohistochemistry and direct immunofluorescent staining of hRPE and McCoy cell cultures at different time-points postinoculation with Chlamydia trachomatis (clinical isolate No.24032).In both types of cultures, both techniques reveal highly immunoreactive inclusions (arrowheads). Twenty-four to 72 h postinoculation, a reduction in the number of intracellular inclusions is observed due to the release of a new generation of EBs to the extracellular environment. Scale bar: 50 μm.
Mentions: In all non-control samples of RPE and McCoy cell cultures, the pathogen was detected both by immunohistochemistry and by direct immunofluorescent staining (Fig 1). Twenty-four hours postinoculation (hpi), clinical isolates (at equal multiplicity of infection (MOI) of 2.0) demonstrated a wide variation in infectivity, with the percentage of RPE culture inclusion-containing cells ranging from 1.5±0.52% (isolate No.3438) to 14.6±3.3% (isolate No.24032), whereas that of McCoy culture cells infected at equal MOI of 2.0 was 0.37±0.34% (isolate No.1609) to 8.9±2.4% (isolate No.24032) (Fig 2). Twenty-four hpi, the variation in the percentage of RPE culture inclusion-containing cells among five clinical isolates used for infection at MOI 2.0 was almost 10-fold (1.5–14.6%). All the five clinical isolates used at a higher MOI for infection of RPE cells than for infection of McCoy cells showed a significantly higher (P<0.05) percentage of inclusion-containing cells for hRPE cell culture than for McCoy cell culture. At equal MOI of 0.3, no statistically significant difference in the percentage of inclusion-containing cells was found between McCoy cells and RPR cells.

Bottom Line: All eight clinical isolates demonstrated ability to infect hRPE cells.At 24h PI, the percentage of inclusion-containing cells varied from 1.5 ± 0.52 to 14.6 ± 3.3% in hRPE cell culture infected at MOI of 2.0 against 0.37 ± 0.34 to 8.9 ± 0.2% in McCoy cell culture infected at MOI of 0.3.Collagen type I, collagen type IV, basic fibroblast growth factor, transforming growth factor-beta and interleukin-8 expression at 48h PI were maximally increased, by 2.1-, 1.3-, 1.5-, 1.5- and 1.6-fold, respectively, in the Chlamydia trachomatis-infected compared with control hRPE cell culture specimens (P < 0.05).

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Military Medical Academy, St Petersburg, Russia.

ABSTRACT

Purpose: Little is known about the susceptibility of posterior segment tissues, particularly the human retinal pigment epithelium (hRPE), to Chlamydia trachomatis. The purpose of the study was to investigate the possibility of infecting the hRPE with Chlamydia trachomatis, and to examine the infectivity of different Chlamydia trachomatis clinical isolates for hRPE cells and the hRPE cell response to the infection.

Methods: Cultured hRPE and McCoy cells were inoculated with eight Chlamydia trachomatis (serovar E) clinical isolates at multiplicity of infection (MOI) of 2.0 or 0.3. To detect Chlamydia trachomatis, samples were stained immunohistochemically with anti-major outer membrane protein antibodies at 24h, 48h, and 72h postinoculation (PI). The changes in the expression of signaling molecules and proteins of cytoskeleton and extracellular matrix in hRPE cells were examined immunohistochemically.

Results: All eight clinical isolates demonstrated ability to infect hRPE cells. At equal MOI of 0.3, the infectivity of Chlamydia trachomatis clinical isolates for RPE culture was found to be at least as high as that for McCoy cell culture. At 24h PI, the percentage of inclusion-containing cells varied from 1.5 ± 0.52 to 14.6 ± 3.3% in hRPE cell culture infected at MOI of 2.0 against 0.37 ± 0.34 to 8.9 ± 0.2% in McCoy cell culture infected at MOI of 0.3. Collagen type I, collagen type IV, basic fibroblast growth factor, transforming growth factor-beta and interleukin-8 expression at 48h PI were maximally increased, by 2.1-, 1.3-, 1.5-, 1.5- and 1.6-fold, respectively, in the Chlamydia trachomatis-infected compared with control hRPE cell culture specimens (P < 0.05).

Conclusions: This study, for the first time, proved the possibility of infecting hRPE cultured cells with Chlamydia trachomatis, which leads to proproliferative and proinflammatory changes in the expression of signaling molecules and extracellular matrix components.

No MeSH data available.


Related in: MedlinePlus