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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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Percentage of keratinocytes expressing CXCL1/2 (a), TNF-α (b), IL-1β (c) and epithelial cells expressing CXCL1/2 (d) in control and HSV-2-infected keratinocyte cell cultures at 18 h p.i. exposed or not to anti-Fas cytotoxic antibody (10 μg/ml) and anti-FasL blocking antibody (10 μg/ml). (e) Transwell migration assay with control and HSV-2-infected keratinocyte cell cultures at 18 h p.i. exposed or not to anti-Fas cytotoxic antibody (10 μg/ml) and anti-FasL blocking antibody (10 μg/ml). (f) Numbers of infected CXCL1/2 and IL-1β positive cells per 100 cells found at infected sites in the HSV-2-infected Fas (−/−), FasL (−/−) and WT (C57BL/6) mice at 3 days of infection (g) Numbers of uninfected CXCL1/2 and IL-1β-positive cells per 100 cells found at infected sites in the HSV-2-infected Fas (−/−), FasL (−/−) and WT (C57BL/6) mice at 3 days of infection. Each bar represents the mean from three experiments (N=3)±S.E.M. *Represents significant differences with P≤0.05, whereas **P≤0.001
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fig7: Percentage of keratinocytes expressing CXCL1/2 (a), TNF-α (b), IL-1β (c) and epithelial cells expressing CXCL1/2 (d) in control and HSV-2-infected keratinocyte cell cultures at 18 h p.i. exposed or not to anti-Fas cytotoxic antibody (10 μg/ml) and anti-FasL blocking antibody (10 μg/ml). (e) Transwell migration assay with control and HSV-2-infected keratinocyte cell cultures at 18 h p.i. exposed or not to anti-Fas cytotoxic antibody (10 μg/ml) and anti-FasL blocking antibody (10 μg/ml). (f) Numbers of infected CXCL1/2 and IL-1β positive cells per 100 cells found at infected sites in the HSV-2-infected Fas (−/−), FasL (−/−) and WT (C57BL/6) mice at 3 days of infection (g) Numbers of uninfected CXCL1/2 and IL-1β-positive cells per 100 cells found at infected sites in the HSV-2-infected Fas (−/−), FasL (−/−) and WT (C57BL/6) mice at 3 days of infection. Each bar represents the mean from three experiments (N=3)±S.E.M. *Represents significant differences with P≤0.05, whereas **P≤0.001

Mentions: To assess whether stimulation through the Fas/FasL pathway influences the induction of inflammatory responses by HSV-2-infected keratinocytes and epithelial cells, we evaluated the expression of CXCL1/2 chemokines, TNF-α and IL-1β cytokines. HSV-2 infection of keratinocytes in vitro led to a significant upregulation of CXCL1/2, TNF-α and IL-1β expression at 18 h of infection (P≤0.001) (Figures 7a–c), whereas HSV-2-infected epithelial cells upregulated CXCL1/2 (Figure 7d), TNF-α and IL-1β (data not shown). Addition of anti-Fas cytotoxic antibody to control uninfected keratinocytes led to a significant upregulation of CXCL1/2 and TNF-α expression at 18 h of incubation (P≤0.001) (Figures 7a and b). Interestingly, addition of the cytotoxic anti-Fas antibody to HSV-2-infected keratinocyte and epithelial cultures significantly abrogated CXCL1/2 expression, whereas, upon these conditions, TNF-α and IL-1β expression was abrogated only in keratinocytes (P≤0.05, Figures 7a–d).


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)

Percentage of keratinocytes expressing CXCL1/2 (a), TNF-α (b), IL-1β (c) and epithelial cells expressing CXCL1/2 (d) in control and HSV-2-infected keratinocyte cell cultures at 18 h p.i. exposed or not to anti-Fas cytotoxic antibody (10 μg/ml) and anti-FasL blocking antibody (10 μg/ml). (e) Transwell migration assay with control and HSV-2-infected keratinocyte cell cultures at 18 h p.i. exposed or not to anti-Fas cytotoxic antibody (10 μg/ml) and anti-FasL blocking antibody (10 μg/ml). (f) Numbers of infected CXCL1/2 and IL-1β positive cells per 100 cells found at infected sites in the HSV-2-infected Fas (−/−), FasL (−/−) and WT (C57BL/6) mice at 3 days of infection (g) Numbers of uninfected CXCL1/2 and IL-1β-positive cells per 100 cells found at infected sites in the HSV-2-infected Fas (−/−), FasL (−/−) and WT (C57BL/6) mice at 3 days of infection. Each bar represents the mean from three experiments (N=3)±S.E.M. *Represents significant differences with P≤0.05, whereas **P≤0.001
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fig7: Percentage of keratinocytes expressing CXCL1/2 (a), TNF-α (b), IL-1β (c) and epithelial cells expressing CXCL1/2 (d) in control and HSV-2-infected keratinocyte cell cultures at 18 h p.i. exposed or not to anti-Fas cytotoxic antibody (10 μg/ml) and anti-FasL blocking antibody (10 μg/ml). (e) Transwell migration assay with control and HSV-2-infected keratinocyte cell cultures at 18 h p.i. exposed or not to anti-Fas cytotoxic antibody (10 μg/ml) and anti-FasL blocking antibody (10 μg/ml). (f) Numbers of infected CXCL1/2 and IL-1β positive cells per 100 cells found at infected sites in the HSV-2-infected Fas (−/−), FasL (−/−) and WT (C57BL/6) mice at 3 days of infection (g) Numbers of uninfected CXCL1/2 and IL-1β-positive cells per 100 cells found at infected sites in the HSV-2-infected Fas (−/−), FasL (−/−) and WT (C57BL/6) mice at 3 days of infection. Each bar represents the mean from three experiments (N=3)±S.E.M. *Represents significant differences with P≤0.05, whereas **P≤0.001
Mentions: To assess whether stimulation through the Fas/FasL pathway influences the induction of inflammatory responses by HSV-2-infected keratinocytes and epithelial cells, we evaluated the expression of CXCL1/2 chemokines, TNF-α and IL-1β cytokines. HSV-2 infection of keratinocytes in vitro led to a significant upregulation of CXCL1/2, TNF-α and IL-1β expression at 18 h of infection (P≤0.001) (Figures 7a–c), whereas HSV-2-infected epithelial cells upregulated CXCL1/2 (Figure 7d), TNF-α and IL-1β (data not shown). Addition of anti-Fas cytotoxic antibody to control uninfected keratinocytes led to a significant upregulation of CXCL1/2 and TNF-α expression at 18 h of incubation (P≤0.001) (Figures 7a and b). Interestingly, addition of the cytotoxic anti-Fas antibody to HSV-2-infected keratinocyte and epithelial cultures significantly abrogated CXCL1/2 expression, whereas, upon these conditions, TNF-α and IL-1β expression was abrogated only in keratinocytes (P≤0.05, Figures 7a–d).

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