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BTN3A2 expression in epithelial ovarian cancer is associated with higher tumor infiltrating T cells and a better prognosis.

Le Page C, Marineau A, Bonza PK, Rahimi K, Cyr L, Labouba I, Madore J, Delvoye N, Mes-Masson AM, Provencher DM, Cailhier JF - PLoS ONE (2012)

Bottom Line: BTN3A2/BT3.2 butyrophilin mRNA expression by tumoral cells was previously identified as a prognostic factor in a small cohort of high grade serous epithelial ovarian cancer (HG-EOC).In conclusion, BT3.2 protein is a potential prognostic biomarker for the identification of HG-EOC patients with better outcome.While the role of BT3.2 is still unknown, our result suggest that BT3.2 expression by epithelial cells may modulates the intratumoral infiltration of immune cells.

View Article: PubMed Central - PubMed

Affiliation: Centre de Recherche du Centre Hospitalier de l'Université de Montréal (CRCHUM), Montreal, Quebec, Canada.

ABSTRACT
BTN3A2/BT3.2 butyrophilin mRNA expression by tumoral cells was previously identified as a prognostic factor in a small cohort of high grade serous epithelial ovarian cancer (HG-EOC). Here, we evaluated the prognostic value of BT3.2 at the protein level in specimen from 199 HG-EOC patients. As the only known role of butyrophilin proteins is in immune regulation, we evaluated the association between BT3.2 expression and intratumoral infiltration of immune cells by immunohistochemistry with specific antibodies against BT3.2, CD3, CD4, CD8, CD20, CD68 and CD206. Epithelial BT3.2 expression was significantly associated with longer overall survival and lower risk of disease progression (HR=0.651, p=0.006 and HR=0.642, p=0.002, respectively) and significantly associated with a higher density of infiltrating T cells, particularly CD4+ cells (0.272, p<0.001). We also observed a strong association between the relative density of CD206+ cells, as evaluated by the ratio of intratumoral CD206+/CD68+ expression, and risk of disease progression (HR=1.355 p=0.044, respectively). In conclusion, BT3.2 protein is a potential prognostic biomarker for the identification of HG-EOC patients with better outcome. In contrast, high CD206+/CD68+ expression is associated with high risk of disease progression. While the role of BT3.2 is still unknown, our result suggest that BT3.2 expression by epithelial cells may modulates the intratumoral infiltration of immune cells.

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BT3.2 expression in ovarian cancer cells.A. Western-blot analysis of total protein extracts from TOV112D cell line infected with PLenti (vector control), BT3.1, BT3.2 or BT3.3 viral constructs. Extracts were loaded in triplicate on 10% SDS/PAGE gel and membranes were hybridized with anti-CD277 (eBioscience), anti-BT3.3 (Atlas Antibodies) and anti-BT3.2 (SDIX Inc.) as indicated at the bottom of the Figure. B. Immunohistochemistry analysis of paraffin-embedded cell pellets from TOV112D cell line infected with either PLenti (vector control), BT3.1, BT3.2 or the BT3.3 viral constructs. Only cells transfected with the BT3.2 construct stained positively with the anti-BT3.2 (SDIX Inc.). C. Immunohistochemistry of xenograft tumors from TOV112D transfected with either empty vector or BT3.2 construct. Only cells infected with the BT3.2 construct stained positively with the anti-BT3.2 (SDIX Inc.). D. Representative staining for immunohistochemistry of BT3.2 on a high-grade serous EOC TMA. From left to right: negative, low, moderate and high intensity.
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pone-0038541-g001: BT3.2 expression in ovarian cancer cells.A. Western-blot analysis of total protein extracts from TOV112D cell line infected with PLenti (vector control), BT3.1, BT3.2 or BT3.3 viral constructs. Extracts were loaded in triplicate on 10% SDS/PAGE gel and membranes were hybridized with anti-CD277 (eBioscience), anti-BT3.3 (Atlas Antibodies) and anti-BT3.2 (SDIX Inc.) as indicated at the bottom of the Figure. B. Immunohistochemistry analysis of paraffin-embedded cell pellets from TOV112D cell line infected with either PLenti (vector control), BT3.1, BT3.2 or the BT3.3 viral constructs. Only cells transfected with the BT3.2 construct stained positively with the anti-BT3.2 (SDIX Inc.). C. Immunohistochemistry of xenograft tumors from TOV112D transfected with either empty vector or BT3.2 construct. Only cells infected with the BT3.2 construct stained positively with the anti-BT3.2 (SDIX Inc.). D. Representative staining for immunohistochemistry of BT3.2 on a high-grade serous EOC TMA. From left to right: negative, low, moderate and high intensity.

Mentions: Tumor sections were scanned and digitally visualized. For BT3.2, epithelial zones were scored according to the staining intensity of the epithelial membrane (value of 0 for absence, 1 for weak, 2 for moderate, 3 for high intensity) as illustrated in Figure 1. In cores where staining was of variable intensity the average intensity was reported. The quantification of infiltrating lymphocytes was carried out by counting the number of nucleated cells with positive staining in intraepithelial islets. Results were then categorized as 0 (no cells), 1 (0<n<10), 2 (10<n<90) and 3 (n>90) accordingly of the number of cells counted. The quantification of macrophages was carried out by evaluating the percentage of stained cells in the intraepithelial area. The relative density of CD206+cells was calculated by the ratio CD206+/CD68+ stained cells. Each array was independently analyzed in a blind study by two independent observers. Inter-rating correlation was >75%. When strong differences in scoring between the two observers (more than 1 unit per core) occurred the core was re-evaluated to reach a concordant scoring between the two observers. The average of all cores with cancer from the same patient was used for analysis.


BTN3A2 expression in epithelial ovarian cancer is associated with higher tumor infiltrating T cells and a better prognosis.

Le Page C, Marineau A, Bonza PK, Rahimi K, Cyr L, Labouba I, Madore J, Delvoye N, Mes-Masson AM, Provencher DM, Cailhier JF - PLoS ONE (2012)

BT3.2 expression in ovarian cancer cells.A. Western-blot analysis of total protein extracts from TOV112D cell line infected with PLenti (vector control), BT3.1, BT3.2 or BT3.3 viral constructs. Extracts were loaded in triplicate on 10% SDS/PAGE gel and membranes were hybridized with anti-CD277 (eBioscience), anti-BT3.3 (Atlas Antibodies) and anti-BT3.2 (SDIX Inc.) as indicated at the bottom of the Figure. B. Immunohistochemistry analysis of paraffin-embedded cell pellets from TOV112D cell line infected with either PLenti (vector control), BT3.1, BT3.2 or the BT3.3 viral constructs. Only cells transfected with the BT3.2 construct stained positively with the anti-BT3.2 (SDIX Inc.). C. Immunohistochemistry of xenograft tumors from TOV112D transfected with either empty vector or BT3.2 construct. Only cells infected with the BT3.2 construct stained positively with the anti-BT3.2 (SDIX Inc.). D. Representative staining for immunohistochemistry of BT3.2 on a high-grade serous EOC TMA. From left to right: negative, low, moderate and high intensity.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3369854&req=5

pone-0038541-g001: BT3.2 expression in ovarian cancer cells.A. Western-blot analysis of total protein extracts from TOV112D cell line infected with PLenti (vector control), BT3.1, BT3.2 or BT3.3 viral constructs. Extracts were loaded in triplicate on 10% SDS/PAGE gel and membranes were hybridized with anti-CD277 (eBioscience), anti-BT3.3 (Atlas Antibodies) and anti-BT3.2 (SDIX Inc.) as indicated at the bottom of the Figure. B. Immunohistochemistry analysis of paraffin-embedded cell pellets from TOV112D cell line infected with either PLenti (vector control), BT3.1, BT3.2 or the BT3.3 viral constructs. Only cells transfected with the BT3.2 construct stained positively with the anti-BT3.2 (SDIX Inc.). C. Immunohistochemistry of xenograft tumors from TOV112D transfected with either empty vector or BT3.2 construct. Only cells infected with the BT3.2 construct stained positively with the anti-BT3.2 (SDIX Inc.). D. Representative staining for immunohistochemistry of BT3.2 on a high-grade serous EOC TMA. From left to right: negative, low, moderate and high intensity.
Mentions: Tumor sections were scanned and digitally visualized. For BT3.2, epithelial zones were scored according to the staining intensity of the epithelial membrane (value of 0 for absence, 1 for weak, 2 for moderate, 3 for high intensity) as illustrated in Figure 1. In cores where staining was of variable intensity the average intensity was reported. The quantification of infiltrating lymphocytes was carried out by counting the number of nucleated cells with positive staining in intraepithelial islets. Results were then categorized as 0 (no cells), 1 (0<n<10), 2 (10<n<90) and 3 (n>90) accordingly of the number of cells counted. The quantification of macrophages was carried out by evaluating the percentage of stained cells in the intraepithelial area. The relative density of CD206+cells was calculated by the ratio CD206+/CD68+ stained cells. Each array was independently analyzed in a blind study by two independent observers. Inter-rating correlation was >75%. When strong differences in scoring between the two observers (more than 1 unit per core) occurred the core was re-evaluated to reach a concordant scoring between the two observers. The average of all cores with cancer from the same patient was used for analysis.

Bottom Line: BTN3A2/BT3.2 butyrophilin mRNA expression by tumoral cells was previously identified as a prognostic factor in a small cohort of high grade serous epithelial ovarian cancer (HG-EOC).In conclusion, BT3.2 protein is a potential prognostic biomarker for the identification of HG-EOC patients with better outcome.While the role of BT3.2 is still unknown, our result suggest that BT3.2 expression by epithelial cells may modulates the intratumoral infiltration of immune cells.

View Article: PubMed Central - PubMed

Affiliation: Centre de Recherche du Centre Hospitalier de l'Université de Montréal (CRCHUM), Montreal, Quebec, Canada.

ABSTRACT
BTN3A2/BT3.2 butyrophilin mRNA expression by tumoral cells was previously identified as a prognostic factor in a small cohort of high grade serous epithelial ovarian cancer (HG-EOC). Here, we evaluated the prognostic value of BT3.2 at the protein level in specimen from 199 HG-EOC patients. As the only known role of butyrophilin proteins is in immune regulation, we evaluated the association between BT3.2 expression and intratumoral infiltration of immune cells by immunohistochemistry with specific antibodies against BT3.2, CD3, CD4, CD8, CD20, CD68 and CD206. Epithelial BT3.2 expression was significantly associated with longer overall survival and lower risk of disease progression (HR=0.651, p=0.006 and HR=0.642, p=0.002, respectively) and significantly associated with a higher density of infiltrating T cells, particularly CD4+ cells (0.272, p<0.001). We also observed a strong association between the relative density of CD206+ cells, as evaluated by the ratio of intratumoral CD206+/CD68+ expression, and risk of disease progression (HR=1.355 p=0.044, respectively). In conclusion, BT3.2 protein is a potential prognostic biomarker for the identification of HG-EOC patients with better outcome. In contrast, high CD206+/CD68+ expression is associated with high risk of disease progression. While the role of BT3.2 is still unknown, our result suggest that BT3.2 expression by epithelial cells may modulates the intratumoral infiltration of immune cells.

Show MeSH
Related in: MedlinePlus