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Role of Fas/FasL in regulation of inflammation in vaginal tissue during HSV-2 infection.

Krzyzowska M, Shestakov A, Eriksson K, Chiodi F - Cell Death Dis (2011)

Bottom Line: Despite upregulation of Fas and FasL, HSV-2-infected keratinocytes and epithelial cells showed a moderate level of apoptosis due to upregulated expression of the anti-apoptotic factors Bcl-2, Akt kinase and NF-κB.Inflammatory lesions within the HSV-2-infected epithelium of C57BL6 mice consisted of infected cells upregulating Fas, FasL and Bcl-2, uninfected cells upregulating Fas and neutrophils expressing both Fas and FasL.We conclude that the Fas pathway participates in regulation of inflammatory response in the vaginal epithelium at the initial stage of HSV-2 infection.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden. krzyzowskam@yahoo.com

ABSTRACT
To assess the role of Fas in lesion development during genital HSV-2 infection, we used a well-established HSV-2 murine model applied to MRL-Fas(lpr)/J (Fas-/-) and C3-Fasl(gld)/J (FasL-/-) C57BL6 mice. In vitro infection of murine keratinocytes and epithelial cells was used to clarify molecular details of HSV-2 infection. Despite upregulation of Fas and FasL, HSV-2-infected keratinocytes and epithelial cells showed a moderate level of apoptosis due to upregulated expression of the anti-apoptotic factors Bcl-2, Akt kinase and NF-κB. Inflammatory lesions within the HSV-2-infected epithelium of C57BL6 mice consisted of infected cells upregulating Fas, FasL and Bcl-2, uninfected cells upregulating Fas and neutrophils expressing both Fas and FasL. Apoptosis was detected in HSV-2-infected cells and to even higher extent in non-infected cells surrounding HSV-2 infection sites. HSV-2 infection of Fas- and FasL-deficient mice led to increased apoptosis and stronger recruitment of neutrophils within the infection sites. We conclude that the Fas pathway participates in regulation of inflammatory response in the vaginal epithelium at the initial stage of HSV-2 infection.

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(a) Kinetics of total percentage of M30-positive (apoptotic) cells in epithelial Hepa 1–6 and keratinocyte 03C cell cultures during 48 h after infection with HSV-2, (b) the total cell number in cultures of HSV-2-infected epithelial cells and keratinocytes (at 24 h p.i.) and in control, uninfected cultures, (c) the total percentage of HSV-2-infected cells in epithelial and keratinocyte cell cultures at 24 and 48 h p.i., (d) double-positive M30/HSV-2 cells in HSV-2-infected epithelial and keratinocyte cultures during 48 h of infection with HSV-2. (e) Percentage of M30-positive cells in epithelial Hepa 1–6 and keratinocyte 03C cell cultures subjected to ST (5 μM), infected or not, with HSV-2 for 18 or 22 h (including 4 h infection, before staurosporine treatment) at MOI=1. Each bar represents the mean of five independent experiments (N=5)±S.E.M.
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fig1: (a) Kinetics of total percentage of M30-positive (apoptotic) cells in epithelial Hepa 1–6 and keratinocyte 03C cell cultures during 48 h after infection with HSV-2, (b) the total cell number in cultures of HSV-2-infected epithelial cells and keratinocytes (at 24 h p.i.) and in control, uninfected cultures, (c) the total percentage of HSV-2-infected cells in epithelial and keratinocyte cell cultures at 24 and 48 h p.i., (d) double-positive M30/HSV-2 cells in HSV-2-infected epithelial and keratinocyte cultures during 48 h of infection with HSV-2. (e) Percentage of M30-positive cells in epithelial Hepa 1–6 and keratinocyte 03C cell cultures subjected to ST (5 μM), infected or not, with HSV-2 for 18 or 22 h (including 4 h infection, before staurosporine treatment) at MOI=1. Each bar represents the mean of five independent experiments (N=5)±S.E.M.

Mentions: A significantly higher number of cells, with caspase-3-specific cytokeratin 18 cleavage (M30-positive cells), was found in HSV-2-infected mouse keratinocytes (291.03C) and epithelial cells (Hepa 1–6) at 24 and 48 h postinfection (p.i.) in comparison with uninfected cultures (P≤0.05) (Figure 1a). Interestingly, in spite of the fact that both infected epithelial and keratinocyte cultures underwent apoptosis, the total cell number in these cultures was comparable to the control cell cultures (Figure 1b).


Role of Fas/FasL in regulation of inflammation in vaginal tissue during HSV-2 infection.

Krzyzowska M, Shestakov A, Eriksson K, Chiodi F - Cell Death Dis (2011)

(a) Kinetics of total percentage of M30-positive (apoptotic) cells in epithelial Hepa 1–6 and keratinocyte 03C cell cultures during 48 h after infection with HSV-2, (b) the total cell number in cultures of HSV-2-infected epithelial cells and keratinocytes (at 24 h p.i.) and in control, uninfected cultures, (c) the total percentage of HSV-2-infected cells in epithelial and keratinocyte cell cultures at 24 and 48 h p.i., (d) double-positive M30/HSV-2 cells in HSV-2-infected epithelial and keratinocyte cultures during 48 h of infection with HSV-2. (e) Percentage of M30-positive cells in epithelial Hepa 1–6 and keratinocyte 03C cell cultures subjected to ST (5 μM), infected or not, with HSV-2 for 18 or 22 h (including 4 h infection, before staurosporine treatment) at MOI=1. Each bar represents the mean of five independent experiments (N=5)±S.E.M.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3101811&req=5

fig1: (a) Kinetics of total percentage of M30-positive (apoptotic) cells in epithelial Hepa 1–6 and keratinocyte 03C cell cultures during 48 h after infection with HSV-2, (b) the total cell number in cultures of HSV-2-infected epithelial cells and keratinocytes (at 24 h p.i.) and in control, uninfected cultures, (c) the total percentage of HSV-2-infected cells in epithelial and keratinocyte cell cultures at 24 and 48 h p.i., (d) double-positive M30/HSV-2 cells in HSV-2-infected epithelial and keratinocyte cultures during 48 h of infection with HSV-2. (e) Percentage of M30-positive cells in epithelial Hepa 1–6 and keratinocyte 03C cell cultures subjected to ST (5 μM), infected or not, with HSV-2 for 18 or 22 h (including 4 h infection, before staurosporine treatment) at MOI=1. Each bar represents the mean of five independent experiments (N=5)±S.E.M.
Mentions: A significantly higher number of cells, with caspase-3-specific cytokeratin 18 cleavage (M30-positive cells), was found in HSV-2-infected mouse keratinocytes (291.03C) and epithelial cells (Hepa 1–6) at 24 and 48 h postinfection (p.i.) in comparison with uninfected cultures (P≤0.05) (Figure 1a). Interestingly, in spite of the fact that both infected epithelial and keratinocyte cultures underwent apoptosis, the total cell number in these cultures was comparable to the control cell cultures (Figure 1b).

Bottom Line: Despite upregulation of Fas and FasL, HSV-2-infected keratinocytes and epithelial cells showed a moderate level of apoptosis due to upregulated expression of the anti-apoptotic factors Bcl-2, Akt kinase and NF-κB.Inflammatory lesions within the HSV-2-infected epithelium of C57BL6 mice consisted of infected cells upregulating Fas, FasL and Bcl-2, uninfected cells upregulating Fas and neutrophils expressing both Fas and FasL.We conclude that the Fas pathway participates in regulation of inflammatory response in the vaginal epithelium at the initial stage of HSV-2 infection.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden. krzyzowskam@yahoo.com

ABSTRACT
To assess the role of Fas in lesion development during genital HSV-2 infection, we used a well-established HSV-2 murine model applied to MRL-Fas(lpr)/J (Fas-/-) and C3-Fasl(gld)/J (FasL-/-) C57BL6 mice. In vitro infection of murine keratinocytes and epithelial cells was used to clarify molecular details of HSV-2 infection. Despite upregulation of Fas and FasL, HSV-2-infected keratinocytes and epithelial cells showed a moderate level of apoptosis due to upregulated expression of the anti-apoptotic factors Bcl-2, Akt kinase and NF-κB. Inflammatory lesions within the HSV-2-infected epithelium of C57BL6 mice consisted of infected cells upregulating Fas, FasL and Bcl-2, uninfected cells upregulating Fas and neutrophils expressing both Fas and FasL. Apoptosis was detected in HSV-2-infected cells and to even higher extent in non-infected cells surrounding HSV-2 infection sites. HSV-2 infection of Fas- and FasL-deficient mice led to increased apoptosis and stronger recruitment of neutrophils within the infection sites. We conclude that the Fas pathway participates in regulation of inflammatory response in the vaginal epithelium at the initial stage of HSV-2 infection.

Show MeSH
Related in: MedlinePlus