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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 expression levels of survivin splice variants in peripheral blood mononuclear cells (PBMCs) from systemic sclerosis (SSc) patients. a The scheme of survivin splice variants. BIRC5 gene could generate several splice variants. b The reverse transcription polymerase chain reaction (RT-PCR) analysis of PBMCs from healthy controls (HCs) (n = 5) and SSc patients with interstitial pneumonia (IP) (n = 5). The figure shown is representative of the samples. c Relative expression levels of survivin (-WT, -ΔEx3 and -2B) in PBMCs from HCs (n = 5) and SSc patients (n = 5) using real-time PCR. *Mann–Whitney U test, p < 0.05. d PBMCs were collected from HCs (n = 4) and SSc patients (n = 3), and fluorescence-activated cell sorting (FACS) analyses were performed for the measurement of intracellular survivin expression. The SSc patients were designated as numbers 1, 3 and 5 in Table 2. Stained PBMCs were gated using the forward and side scatters and CD45+CD14+ cells were analyzed (gray line, normal immunoglobulin G (IgG) control; green line, HC; red line, SSc). Representative images and their mean fluorescence intensity (MFI) are shown. e Estimated survivin-wild type (WT) expression levels of peripheral CD45+CD14+ cells in HCs (n = 4) and SSc patients (n = 3). Mean ± SEM, *t test, p < 0.05
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Fig3: The expression levels of survivin splice variants in peripheral blood mononuclear cells (PBMCs) from systemic sclerosis (SSc) patients. a The scheme of survivin splice variants. BIRC5 gene could generate several splice variants. b The reverse transcription polymerase chain reaction (RT-PCR) analysis of PBMCs from healthy controls (HCs) (n = 5) and SSc patients with interstitial pneumonia (IP) (n = 5). The figure shown is representative of the samples. c Relative expression levels of survivin (-WT, -ΔEx3 and -2B) in PBMCs from HCs (n = 5) and SSc patients (n = 5) using real-time PCR. *Mann–Whitney U test, p < 0.05. d PBMCs were collected from HCs (n = 4) and SSc patients (n = 3), and fluorescence-activated cell sorting (FACS) analyses were performed for the measurement of intracellular survivin expression. The SSc patients were designated as numbers 1, 3 and 5 in Table 2. Stained PBMCs were gated using the forward and side scatters and CD45+CD14+ cells were analyzed (gray line, normal immunoglobulin G (IgG) control; green line, HC; red line, SSc). Representative images and their mean fluorescence intensity (MFI) are shown. e Estimated survivin-wild type (WT) expression levels of peripheral CD45+CD14+ cells in HCs (n = 4) and SSc patients (n = 3). Mean ± SEM, *t test, p < 0.05

Mentions: It was reported that the BIRC5 gene could generate survivin splice variants, which result from alternative splicing (Fig. 3a) [20]. RT-PCR revealed that the expressions of survivin-WT, -ΔEx3 and -2B were detectable in PBMCs from SSc patients (n = 5) and HCs (n = 5) (Fig. 3b). Subsequently, we performed quantification of the survivin splicing variants using real-time PCR. As a result, the expression levels of only survivin-WT in PBMCs from SSc patients were higher than those from the controls (p = 0.016) (Fig. 3c). There were no significant differences in the expression levels of survivin-ΔEx3 or -2B. Moreover, we also measured the intracellular expression levels of survivin proteins in PBMCs using flow cytometry. As shown in Fig. 3d, flow cytometry using anti-survivin antibody, which can detect all survivin variants (Fig. 2e), revealed that the survivin expression levels in monocytes from SSc patients (CD45+CD14+ cells) were higher than those from HCs (Fig. 3d). However, the expression levels of survivin-ΔEx3 and -2B in CD45+CD14+ cells were similar between SSc patients and HCs. The estimated survivin-WT expression levels of CD45+CD14+ cells in SSc patients (n = 3) were higher than those in HCs (n = 4) (p = 0.043) (Fig. 3e).Fig. 3


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 expression levels of survivin splice variants in peripheral blood mononuclear cells (PBMCs) from systemic sclerosis (SSc) patients. a The scheme of survivin splice variants. BIRC5 gene could generate several splice variants. b The reverse transcription polymerase chain reaction (RT-PCR) analysis of PBMCs from healthy controls (HCs) (n = 5) and SSc patients with interstitial pneumonia (IP) (n = 5). The figure shown is representative of the samples. c Relative expression levels of survivin (-WT, -ΔEx3 and -2B) in PBMCs from HCs (n = 5) and SSc patients (n = 5) using real-time PCR. *Mann–Whitney U test, p < 0.05. d PBMCs were collected from HCs (n = 4) and SSc patients (n = 3), and fluorescence-activated cell sorting (FACS) analyses were performed for the measurement of intracellular survivin expression. The SSc patients were designated as numbers 1, 3 and 5 in Table 2. Stained PBMCs were gated using the forward and side scatters and CD45+CD14+ cells were analyzed (gray line, normal immunoglobulin G (IgG) control; green line, HC; red line, SSc). Representative images and their mean fluorescence intensity (MFI) are shown. e Estimated survivin-wild type (WT) expression levels of peripheral CD45+CD14+ cells in HCs (n = 4) and SSc patients (n = 3). Mean ± SEM, *t test, p < 0.05
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Fig3: The expression levels of survivin splice variants in peripheral blood mononuclear cells (PBMCs) from systemic sclerosis (SSc) patients. a The scheme of survivin splice variants. BIRC5 gene could generate several splice variants. b The reverse transcription polymerase chain reaction (RT-PCR) analysis of PBMCs from healthy controls (HCs) (n = 5) and SSc patients with interstitial pneumonia (IP) (n = 5). The figure shown is representative of the samples. c Relative expression levels of survivin (-WT, -ΔEx3 and -2B) in PBMCs from HCs (n = 5) and SSc patients (n = 5) using real-time PCR. *Mann–Whitney U test, p < 0.05. d PBMCs were collected from HCs (n = 4) and SSc patients (n = 3), and fluorescence-activated cell sorting (FACS) analyses were performed for the measurement of intracellular survivin expression. The SSc patients were designated as numbers 1, 3 and 5 in Table 2. Stained PBMCs were gated using the forward and side scatters and CD45+CD14+ cells were analyzed (gray line, normal immunoglobulin G (IgG) control; green line, HC; red line, SSc). Representative images and their mean fluorescence intensity (MFI) are shown. e Estimated survivin-wild type (WT) expression levels of peripheral CD45+CD14+ cells in HCs (n = 4) and SSc patients (n = 3). Mean ± SEM, *t test, p < 0.05
Mentions: It was reported that the BIRC5 gene could generate survivin splice variants, which result from alternative splicing (Fig. 3a) [20]. RT-PCR revealed that the expressions of survivin-WT, -ΔEx3 and -2B were detectable in PBMCs from SSc patients (n = 5) and HCs (n = 5) (Fig. 3b). Subsequently, we performed quantification of the survivin splicing variants using real-time PCR. As a result, the expression levels of only survivin-WT in PBMCs from SSc patients were higher than those from the controls (p = 0.016) (Fig. 3c). There were no significant differences in the expression levels of survivin-ΔEx3 or -2B. Moreover, we also measured the intracellular expression levels of survivin proteins in PBMCs using flow cytometry. As shown in Fig. 3d, flow cytometry using anti-survivin antibody, which can detect all survivin variants (Fig. 2e), revealed that the survivin expression levels in monocytes from SSc patients (CD45+CD14+ cells) were higher than those from HCs (Fig. 3d). However, the expression levels of survivin-ΔEx3 and -2B in CD45+CD14+ cells were similar between SSc patients and HCs. The estimated survivin-WT expression levels of CD45+CD14+ cells in SSc patients (n = 3) were higher than those in HCs (n = 4) (p = 0.043) (Fig. 3e).Fig. 3

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