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Effect of 12-O-tetradecanoylphorbol-13-acetate-induced psoriasis-like skin lesions on systemic inflammation and atherosclerosis in hypercholesterolaemic apolipoprotein E deficient mice.

Madsen M, Hansen PR, Nielsen LB, Hartvigsen K, Pedersen AE, Christensen JP, Aarup A, Pedersen TX - BMC Dermatol. (2016)

Bottom Line: Systemic effects of the topical application of TPA were demonstrated by increased plasma concentration of serum amyloid A and splenic immune modulation, respectively.TPA-induced psoriasis-like skin inflammation in atherosclerosis-prone ApoE(-/-) mice evoked systemic immune-inflammatory effects, but did not affect atherogenesis.The results may question the role of psoriasis-induced inflammation in the pathogenesis of atherosclerosis in psoriasis patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical Sciences, University of Copenhagen, Copenhagen, Denmark.

ABSTRACT

Background: Risk of cardiovascular disease is increased in patients with psoriasis, but molecular mechanisms linking the two conditions have not been clearly established. Lack of appropriate animal models has hampered generation of new knowledge in this area of research and we therefore sought to develop an animal model with combined atherosclerosis and psoriasis-like skin inflammation.

Methods: Topical 12-O-tetradecanoylphorbol-13-acetate (TPA) was applied to the ears twice per week for 8 weeks in atherosclerosis-prone apolipoprotein E deficient (ApoE(-/-)) mice.

Results: TPA led to localized skin inflammation with increased epidermal thickness, infiltration of inflammatory-like cells and augmented tissue interleukin-17F levels. Systemic effects of the topical application of TPA were demonstrated by increased plasma concentration of serum amyloid A and splenic immune modulation, respectively. However, atherosclerotic plaque area and composition, and mRNA levels of several inflammatory genes in the aortic wall were not significantly affected by TPA-induced skin inflammation.

Conclusions: TPA-induced psoriasis-like skin inflammation in atherosclerosis-prone ApoE(-/-) mice evoked systemic immune-inflammatory effects, but did not affect atherogenesis. The results may question the role of psoriasis-induced inflammation in the pathogenesis of atherosclerosis in psoriasis patients.

No MeSH data available.


Related in: MedlinePlus

Topical 12-O-tetradecanoylphorbol-13-acetate (TPA) application induces systemic inflammation. a Serum amyloid A (SAA) levels (μg/ml) measured in plasma after 8 weeks of topical application on both ears with either TPA or vehicle (control). Note: the y-axis is displayed as a log10 scale. Values are depicted as median and statistical differences analysed by non-parametric t-test. b-f Spleen flow cytometry data. Symbols and horizontal bars indicate individual mice and group averages, respectively, and parametric t-tests were used to detect statistical differences. b Levels of CD11b+ cells, T-cells (CD3+), and B-cells (B220+). c Effector and memory CD4+ T-cells (CD44+CD62L− and CD44+CD62L+ cells, respectively), and d memory CD8+ T-cells (CD44+CD62L+ cells). e Tc1 cells (IFNγ+IL17−). f Regulatory T-cells (Foxp3+CD25−). Unfilled and filled triangles represent control and TPA mice, respectively, from study 2. N = 14–15 mice/group; one control mouse was omitted from flow cytometry data due to abnormal values (i.e., above 3xSD)
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Fig2: Topical 12-O-tetradecanoylphorbol-13-acetate (TPA) application induces systemic inflammation. a Serum amyloid A (SAA) levels (μg/ml) measured in plasma after 8 weeks of topical application on both ears with either TPA or vehicle (control). Note: the y-axis is displayed as a log10 scale. Values are depicted as median and statistical differences analysed by non-parametric t-test. b-f Spleen flow cytometry data. Symbols and horizontal bars indicate individual mice and group averages, respectively, and parametric t-tests were used to detect statistical differences. b Levels of CD11b+ cells, T-cells (CD3+), and B-cells (B220+). c Effector and memory CD4+ T-cells (CD44+CD62L− and CD44+CD62L+ cells, respectively), and d memory CD8+ T-cells (CD44+CD62L+ cells). e Tc1 cells (IFNγ+IL17−). f Regulatory T-cells (Foxp3+CD25−). Unfilled and filled triangles represent control and TPA mice, respectively, from study 2. N = 14–15 mice/group; one control mouse was omitted from flow cytometry data due to abnormal values (i.e., above 3xSD)

Mentions: To investigate whether topical application of TPA would induce not only a local immune response, but also systemic effects, we measured plasma levels of SAA and inflammatory cytokines (IL-1β, −2, −4, −5, −6, −10, and -12p70, and IFNγ, and TNFα), and performed flow cytometry of spleens from TPA and control mice. Plasma SAA levels were higher in TPA-treated vs. control mice (4.1 [3.1–6.7] μg/ml vs. 2.8 [2.7–3.0] μg/ml, p < 0.0001, Fig. 2a), whereas the other measured cytokines either were below ELISA detection limits, or showed no difference between the two groups (data not shown). TPA application caused larger spleens compared to vehicle application (5.4 ± 0.2 vs. 4.6 ± 0.2 mg wet weight/body weight, p = 0.0039); however, this difference was not reflected in absolute splenocyte numbers (105 ± 8 × 106 vs. 98 ± 7 × 106 cells, p > 0.05). Flow cytometry analyses revealed a significantly higher amount of CD11b+ cells in spleens from TPA-treated mice compared to control mice (12.0 ± 1.2 vs. 7.9 ± 0.7 × 106 cells, p = 0.009, Fig. 2b). In mouse spleen, CD11b is expressed primarily by inflammatory monocytes, macrophages, neutrophils, and some subpopulations of dendritic cells [22]. Additional flow cytometry analyses of the spleens did not reveal differences in cytotoxic (CD8+) or helper (CD4+) T-cell populations (data not shown). However, detailed analyses of activated CD4+ and CD8+ T-cell populations, based on expression pattern of CD62L and CD44, revealed significantly expanded effector (CD44+CD62L−) and memory (CD44+CD62L+) CD4+ T-cell populations in the TPA mice compared to control mice (Fig. 2c). In addition, the memory (CD44+CD62L+) CD8+ T-cell population was also expanded in the TPA mice (Fig. 2d). There were corresponding reductions of naïve CD4+ and CD8+ T-cell populations (data not shown). Using TPA/ionomycin-stimulation of splenocytes, we detected similar expression of intracellular IFN-γ, IL-4, and IL-17 (Th1, Th2, and Th17 signature cytokines, respectively) in CD4+ cells from TPA and control mice (data not shown). However, in the CD8+ cells, we found a significantly higher percentage of IFN-γ expression (Tc1-cells) in TPA mice (Fig. 2e). There was a corresponding reduction of uncommitted CD8+ cells, but no differences in IL-4 and IL-17 expression in the CD8+ cells (data not shown). In a separate analysis, we found significantly elevated percentages of splenic CD4+Foxp3+CD25− regulatory T-cells (Tregs) in TPA mice compared to control mice (Fig. 2f). There were no differences in natural CD4+Foxp3+CD25+ Tregs or activated CD4+CD25+Foxp3− T-cells (data not shown).Fig. 2


Effect of 12-O-tetradecanoylphorbol-13-acetate-induced psoriasis-like skin lesions on systemic inflammation and atherosclerosis in hypercholesterolaemic apolipoprotein E deficient mice.

Madsen M, Hansen PR, Nielsen LB, Hartvigsen K, Pedersen AE, Christensen JP, Aarup A, Pedersen TX - BMC Dermatol. (2016)

Topical 12-O-tetradecanoylphorbol-13-acetate (TPA) application induces systemic inflammation. a Serum amyloid A (SAA) levels (μg/ml) measured in plasma after 8 weeks of topical application on both ears with either TPA or vehicle (control). Note: the y-axis is displayed as a log10 scale. Values are depicted as median and statistical differences analysed by non-parametric t-test. b-f Spleen flow cytometry data. Symbols and horizontal bars indicate individual mice and group averages, respectively, and parametric t-tests were used to detect statistical differences. b Levels of CD11b+ cells, T-cells (CD3+), and B-cells (B220+). c Effector and memory CD4+ T-cells (CD44+CD62L− and CD44+CD62L+ cells, respectively), and d memory CD8+ T-cells (CD44+CD62L+ cells). e Tc1 cells (IFNγ+IL17−). f Regulatory T-cells (Foxp3+CD25−). Unfilled and filled triangles represent control and TPA mice, respectively, from study 2. N = 14–15 mice/group; one control mouse was omitted from flow cytometry data due to abnormal values (i.e., above 3xSD)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4940745&req=5

Fig2: Topical 12-O-tetradecanoylphorbol-13-acetate (TPA) application induces systemic inflammation. a Serum amyloid A (SAA) levels (μg/ml) measured in plasma after 8 weeks of topical application on both ears with either TPA or vehicle (control). Note: the y-axis is displayed as a log10 scale. Values are depicted as median and statistical differences analysed by non-parametric t-test. b-f Spleen flow cytometry data. Symbols and horizontal bars indicate individual mice and group averages, respectively, and parametric t-tests were used to detect statistical differences. b Levels of CD11b+ cells, T-cells (CD3+), and B-cells (B220+). c Effector and memory CD4+ T-cells (CD44+CD62L− and CD44+CD62L+ cells, respectively), and d memory CD8+ T-cells (CD44+CD62L+ cells). e Tc1 cells (IFNγ+IL17−). f Regulatory T-cells (Foxp3+CD25−). Unfilled and filled triangles represent control and TPA mice, respectively, from study 2. N = 14–15 mice/group; one control mouse was omitted from flow cytometry data due to abnormal values (i.e., above 3xSD)
Mentions: To investigate whether topical application of TPA would induce not only a local immune response, but also systemic effects, we measured plasma levels of SAA and inflammatory cytokines (IL-1β, −2, −4, −5, −6, −10, and -12p70, and IFNγ, and TNFα), and performed flow cytometry of spleens from TPA and control mice. Plasma SAA levels were higher in TPA-treated vs. control mice (4.1 [3.1–6.7] μg/ml vs. 2.8 [2.7–3.0] μg/ml, p < 0.0001, Fig. 2a), whereas the other measured cytokines either were below ELISA detection limits, or showed no difference between the two groups (data not shown). TPA application caused larger spleens compared to vehicle application (5.4 ± 0.2 vs. 4.6 ± 0.2 mg wet weight/body weight, p = 0.0039); however, this difference was not reflected in absolute splenocyte numbers (105 ± 8 × 106 vs. 98 ± 7 × 106 cells, p > 0.05). Flow cytometry analyses revealed a significantly higher amount of CD11b+ cells in spleens from TPA-treated mice compared to control mice (12.0 ± 1.2 vs. 7.9 ± 0.7 × 106 cells, p = 0.009, Fig. 2b). In mouse spleen, CD11b is expressed primarily by inflammatory monocytes, macrophages, neutrophils, and some subpopulations of dendritic cells [22]. Additional flow cytometry analyses of the spleens did not reveal differences in cytotoxic (CD8+) or helper (CD4+) T-cell populations (data not shown). However, detailed analyses of activated CD4+ and CD8+ T-cell populations, based on expression pattern of CD62L and CD44, revealed significantly expanded effector (CD44+CD62L−) and memory (CD44+CD62L+) CD4+ T-cell populations in the TPA mice compared to control mice (Fig. 2c). In addition, the memory (CD44+CD62L+) CD8+ T-cell population was also expanded in the TPA mice (Fig. 2d). There were corresponding reductions of naïve CD4+ and CD8+ T-cell populations (data not shown). Using TPA/ionomycin-stimulation of splenocytes, we detected similar expression of intracellular IFN-γ, IL-4, and IL-17 (Th1, Th2, and Th17 signature cytokines, respectively) in CD4+ cells from TPA and control mice (data not shown). However, in the CD8+ cells, we found a significantly higher percentage of IFN-γ expression (Tc1-cells) in TPA mice (Fig. 2e). There was a corresponding reduction of uncommitted CD8+ cells, but no differences in IL-4 and IL-17 expression in the CD8+ cells (data not shown). In a separate analysis, we found significantly elevated percentages of splenic CD4+Foxp3+CD25− regulatory T-cells (Tregs) in TPA mice compared to control mice (Fig. 2f). There were no differences in natural CD4+Foxp3+CD25+ Tregs or activated CD4+CD25+Foxp3− T-cells (data not shown).Fig. 2

Bottom Line: Systemic effects of the topical application of TPA were demonstrated by increased plasma concentration of serum amyloid A and splenic immune modulation, respectively.TPA-induced psoriasis-like skin inflammation in atherosclerosis-prone ApoE(-/-) mice evoked systemic immune-inflammatory effects, but did not affect atherogenesis.The results may question the role of psoriasis-induced inflammation in the pathogenesis of atherosclerosis in psoriasis patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical Sciences, University of Copenhagen, Copenhagen, Denmark.

ABSTRACT

Background: Risk of cardiovascular disease is increased in patients with psoriasis, but molecular mechanisms linking the two conditions have not been clearly established. Lack of appropriate animal models has hampered generation of new knowledge in this area of research and we therefore sought to develop an animal model with combined atherosclerosis and psoriasis-like skin inflammation.

Methods: Topical 12-O-tetradecanoylphorbol-13-acetate (TPA) was applied to the ears twice per week for 8 weeks in atherosclerosis-prone apolipoprotein E deficient (ApoE(-/-)) mice.

Results: TPA led to localized skin inflammation with increased epidermal thickness, infiltration of inflammatory-like cells and augmented tissue interleukin-17F levels. Systemic effects of the topical application of TPA were demonstrated by increased plasma concentration of serum amyloid A and splenic immune modulation, respectively. However, atherosclerotic plaque area and composition, and mRNA levels of several inflammatory genes in the aortic wall were not significantly affected by TPA-induced skin inflammation.

Conclusions: TPA-induced psoriasis-like skin inflammation in atherosclerosis-prone ApoE(-/-) mice evoked systemic immune-inflammatory effects, but did not affect atherogenesis. The results may question the role of psoriasis-induced inflammation in the pathogenesis of atherosclerosis in psoriasis patients.

No MeSH data available.


Related in: MedlinePlus