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CD1a+ survivin+ dendritic cell infiltration in dermal lesions of systemic sclerosis.

Mokuda S, Miyazaki T, Ubara Y, Kanno M, Sugiyama E, Takasugi K, Masumoto J - Arthritis Res. Ther. (2015)

Bottom Line: PBMCs and monocytes from SSc patients also overexpressed survivin; therefore, dermal survivin+ DC may be derived from peripheral blood monocytes.Additionally, survivin may be involved in dermal CD1a+ DC proliferation through cell cycle activation and resistance to apoptosis.Survivin may be an important molecule for the pathogenesis of SSc.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima, 734-8551, Japan. sho-mokuda@hiroshima-u.ac.jp.

ABSTRACT

Introduction: Proto-oncogene survivin is a member of the inhibitor of apoptosis (IAP) family of proteins. The presence of serous antibodies against survivin in patients with systemic sclerosis has been previously reported; however, there are few reports regarding the pathophysiological relationship between survivin and systemic sclerosis. We herein investigated the expression and function of survivin in SSc patients.

Methods: We performed immunohistochemistry analyses to determine the expression of XIAP, cIAP and survivin in skin lesions from patients with SSc and non-SSc. The expression levels of survivin in peripheral blood mononuclear cells (PBMCs) obtained from SSc patients and healthy controls were evaluated using RT-PCR and flow cytometry. Additionally, the function of survivin was verified with overexpression experiments using monocyte-derived dendritic cells (Mo-DCs).

Results: The expression patterns of both XIAP and cIAP were similar, while only the survivin expression differed between the SSc and non-SSc skin lesions. Survivin-overexpressing cells were detected in the SSc dermis frequently. The positive rate of survivin in SSc dermis (64.3%, 9/14) was higher than that in non-SSc dermis (11.2%, 1/9). Furthermore, survivin+ cells expressed CD1a, one of the DC markers. Real-time PCR and FACS analyses revealed that the survivin-WT (wild type) expression levels in PBMCs, in particular CD14+ monocytes, from SSc patients were higher than that from healthy controls. Additionally, the overexpression experiments showed that survivin-WT-overexpressing CD1a+ Mo-DCs have the characteristics of promoting cell cycle progression and decreasing apoptotic cells.

Conclusions: These findings suggest that dermal survivin+ CD1a+ cell infiltration may be a potential biomarker of SSc skin lesions. PBMCs and monocytes from SSc patients also overexpressed survivin; therefore, dermal survivin+ DC may be derived from peripheral blood monocytes. Additionally, survivin may be involved in dermal CD1a+ DC proliferation through cell cycle activation and resistance to apoptosis. Survivin may be an important molecule for the pathogenesis of SSc.

No MeSH data available.


Related in: MedlinePlus

The role of survivin in CD1a+ monocyte-derived dendritic cells (Mo-DCs). a CD14+ monocytes change into CD1a+CD14− DCs by culturing with granulocyte macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4. b A Western blot analysis showed that BIRC5 mRNA expressed the survivin-WT protein in Mo-DCs. c Mo-DCs were transfected with survivin-wild type (WT) and mock at day 0, and were subsequently stimulated with GM-CSF and IL-4. At 25 and 50 hours, the samples were analyzed using fluorescence-activated cell sorting (FACS) analyses. The percentages in the figures indicate the ratio of CD1a+CD14− cells. d-f Mo-DCs were transfected with survivin-WT and mock at day 6, and the following FACS analyses were performed at 24 hours. d Harvested Mo-DCs were stained with FVD520 and anti-Ki-67 antibodies. The positive rate was calculated as the number of Ki-67-positive cells among FVD520-negative (living) cells. Data are presented as the percentage of control cells. Mean ± SEM. n = 4 (both groups). *t test, p < 0.05. e, f Harvested Mo-DCs were stained with 7-amino-actinomycinD (7-AAD). The percentages of G2/M phase cells (e) and apoptotic cells (f) were calculated. Data are presented as the percentage of control cells. Mean ± SEM, n = 5 (both groups). t test, *p < 0.05, **p < 0.01
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Fig4: The role of survivin in CD1a+ monocyte-derived dendritic cells (Mo-DCs). a CD14+ monocytes change into CD1a+CD14− DCs by culturing with granulocyte macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4. b A Western blot analysis showed that BIRC5 mRNA expressed the survivin-WT protein in Mo-DCs. c Mo-DCs were transfected with survivin-wild type (WT) and mock at day 0, and were subsequently stimulated with GM-CSF and IL-4. At 25 and 50 hours, the samples were analyzed using fluorescence-activated cell sorting (FACS) analyses. The percentages in the figures indicate the ratio of CD1a+CD14− cells. d-f Mo-DCs were transfected with survivin-WT and mock at day 6, and the following FACS analyses were performed at 24 hours. d Harvested Mo-DCs were stained with FVD520 and anti-Ki-67 antibodies. The positive rate was calculated as the number of Ki-67-positive cells among FVD520-negative (living) cells. Data are presented as the percentage of control cells. Mean ± SEM. n = 4 (both groups). *t test, p < 0.05. e, f Harvested Mo-DCs were stained with 7-amino-actinomycinD (7-AAD). The percentages of G2/M phase cells (e) and apoptotic cells (f) were calculated. Data are presented as the percentage of control cells. Mean ± SEM, n = 5 (both groups). t test, *p < 0.05, **p < 0.01

Mentions: CD1a+ DCs can be generated from PBMCs by culturing in vitro [21] (Fig. 4a). We obtained Mo-DCs from healthy donors, established a transfection method (Fig. 4b), and investigated the function of overexpressed survivin. We selected survivin-WT among the splice variants, according to the results from PBMCs and IHC studies (as mentioned above). When Mo-DCs were transfected to overexpress survivin at day 0 (Fig. 4c), survivin overexpression did not affect the differentiation of DCs until 50 hours.Fig. 4


CD1a+ survivin+ dendritic cell infiltration in dermal lesions of systemic sclerosis.

Mokuda S, Miyazaki T, Ubara Y, Kanno M, Sugiyama E, Takasugi K, Masumoto J - Arthritis Res. Ther. (2015)

The role of survivin in CD1a+ monocyte-derived dendritic cells (Mo-DCs). a CD14+ monocytes change into CD1a+CD14− DCs by culturing with granulocyte macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4. b A Western blot analysis showed that BIRC5 mRNA expressed the survivin-WT protein in Mo-DCs. c Mo-DCs were transfected with survivin-wild type (WT) and mock at day 0, and were subsequently stimulated with GM-CSF and IL-4. At 25 and 50 hours, the samples were analyzed using fluorescence-activated cell sorting (FACS) analyses. The percentages in the figures indicate the ratio of CD1a+CD14− cells. d-f Mo-DCs were transfected with survivin-WT and mock at day 6, and the following FACS analyses were performed at 24 hours. d Harvested Mo-DCs were stained with FVD520 and anti-Ki-67 antibodies. The positive rate was calculated as the number of Ki-67-positive cells among FVD520-negative (living) cells. Data are presented as the percentage of control cells. Mean ± SEM. n = 4 (both groups). *t test, p < 0.05. e, f Harvested Mo-DCs were stained with 7-amino-actinomycinD (7-AAD). The percentages of G2/M phase cells (e) and apoptotic cells (f) were calculated. Data are presented as the percentage of control cells. Mean ± SEM, n = 5 (both groups). t test, *p < 0.05, **p < 0.01
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Fig4: The role of survivin in CD1a+ monocyte-derived dendritic cells (Mo-DCs). a CD14+ monocytes change into CD1a+CD14− DCs by culturing with granulocyte macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4. b A Western blot analysis showed that BIRC5 mRNA expressed the survivin-WT protein in Mo-DCs. c Mo-DCs were transfected with survivin-wild type (WT) and mock at day 0, and were subsequently stimulated with GM-CSF and IL-4. At 25 and 50 hours, the samples were analyzed using fluorescence-activated cell sorting (FACS) analyses. The percentages in the figures indicate the ratio of CD1a+CD14− cells. d-f Mo-DCs were transfected with survivin-WT and mock at day 6, and the following FACS analyses were performed at 24 hours. d Harvested Mo-DCs were stained with FVD520 and anti-Ki-67 antibodies. The positive rate was calculated as the number of Ki-67-positive cells among FVD520-negative (living) cells. Data are presented as the percentage of control cells. Mean ± SEM. n = 4 (both groups). *t test, p < 0.05. e, f Harvested Mo-DCs were stained with 7-amino-actinomycinD (7-AAD). The percentages of G2/M phase cells (e) and apoptotic cells (f) were calculated. Data are presented as the percentage of control cells. Mean ± SEM, n = 5 (both groups). t test, *p < 0.05, **p < 0.01
Mentions: CD1a+ DCs can be generated from PBMCs by culturing in vitro [21] (Fig. 4a). We obtained Mo-DCs from healthy donors, established a transfection method (Fig. 4b), and investigated the function of overexpressed survivin. We selected survivin-WT among the splice variants, according to the results from PBMCs and IHC studies (as mentioned above). When Mo-DCs were transfected to overexpress survivin at day 0 (Fig. 4c), survivin overexpression did not affect the differentiation of DCs until 50 hours.Fig. 4

Bottom Line: PBMCs and monocytes from SSc patients also overexpressed survivin; therefore, dermal survivin+ DC may be derived from peripheral blood monocytes.Additionally, survivin may be involved in dermal CD1a+ DC proliferation through cell cycle activation and resistance to apoptosis.Survivin may be an important molecule for the pathogenesis of SSc.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima, 734-8551, Japan. sho-mokuda@hiroshima-u.ac.jp.

ABSTRACT

Introduction: Proto-oncogene survivin is a member of the inhibitor of apoptosis (IAP) family of proteins. The presence of serous antibodies against survivin in patients with systemic sclerosis has been previously reported; however, there are few reports regarding the pathophysiological relationship between survivin and systemic sclerosis. We herein investigated the expression and function of survivin in SSc patients.

Methods: We performed immunohistochemistry analyses to determine the expression of XIAP, cIAP and survivin in skin lesions from patients with SSc and non-SSc. The expression levels of survivin in peripheral blood mononuclear cells (PBMCs) obtained from SSc patients and healthy controls were evaluated using RT-PCR and flow cytometry. Additionally, the function of survivin was verified with overexpression experiments using monocyte-derived dendritic cells (Mo-DCs).

Results: The expression patterns of both XIAP and cIAP were similar, while only the survivin expression differed between the SSc and non-SSc skin lesions. Survivin-overexpressing cells were detected in the SSc dermis frequently. The positive rate of survivin in SSc dermis (64.3%, 9/14) was higher than that in non-SSc dermis (11.2%, 1/9). Furthermore, survivin+ cells expressed CD1a, one of the DC markers. Real-time PCR and FACS analyses revealed that the survivin-WT (wild type) expression levels in PBMCs, in particular CD14+ monocytes, from SSc patients were higher than that from healthy controls. Additionally, the overexpression experiments showed that survivin-WT-overexpressing CD1a+ Mo-DCs have the characteristics of promoting cell cycle progression and decreasing apoptotic cells.

Conclusions: These findings suggest that dermal survivin+ CD1a+ cell infiltration may be a potential biomarker of SSc skin lesions. PBMCs and monocytes from SSc patients also overexpressed survivin; therefore, dermal survivin+ DC may be derived from peripheral blood monocytes. Additionally, survivin may be involved in dermal CD1a+ DC proliferation through cell cycle activation and resistance to apoptosis. Survivin may be an important molecule for the pathogenesis of SSc.

No MeSH data available.


Related in: MedlinePlus