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Tumor-associated macrophages in oral premalignant lesions coexpress CD163 and STAT1 in a Th1-dominated microenvironment.

Mori K, Haraguchi S, Hiori M, Shimada J, Ohmori Y - BMC Cancer (2015)

Bottom Line: Although CCR4(+) cells rarely infiltrated, CXCR3(+) and CCR5(+) cells were observed in these lesions.Cells positive for STAT1 and chemokine CXCL9, interferon- (IFN)-induced gene products, and pSTAT1 were also observed in the same lesions.Double immunofluorescence staining demonstrated that the cells that were positive for CD163 were also positive for STAT1.

View Article: PubMed Central - PubMed

Affiliation: Division of Oral and Maxillofacial Surgery, Department of Diagnosis and Therapeutics Sciences, Meikai University of School of Dentistry, 1-1 Keyakidai, Sakado, Saitama, 350-0283, Japan. kazu-mori@dent.meikai.ac.jp.

ABSTRACT

Background: Tumor-associated macrophages (TAMs) are implicated in the growth, invasion and metastasis of various solid tumors. However, the phenotype of TAMs in premalignant lesions of solid tumors has not been clarified. In the present study, we identify the phenotype of TAMs in leukoplakia, an oral premalignant lesion, by immunohistochemical analysis and investigate the involvement of infiltrated T cells that participate in the polarization of TAMs.

Methods: The subjects included 30 patients with oral leukoplakia and 10 individuals with normal mucosa. Hematoxylin and eosin slides were examined for the histological grades, and immunohistochemical analysis was carried out using antibodies against CD68 (pan-MΦ), CD80 (M1 MΦ), CD163 (M2 MΦ), CD4 (helper T cells: Th), CD8 (cytotoxic T cells), CXCR3, CCR5 (Th1), CCR4 (Th2), signal transducer and activator of transcription (STAT1), phosphorylated STAT1 (pSTAT1) and chemokine CXCL9. The differences in the numbers of positively stained cells among the different histological grades were tested for statistical significance using the Kruskal-Wallis test. Correlations between different types of immune cells were determined using Spearman's rank analysis.

Results: An increase in the rate of CD163(+) TAM infiltration was observed in mild and moderate epithelial dysplasia, which positively correlated with the rate of intraepithelial CD4(+) Th cell infiltration. Although CCR4(+) cells rarely infiltrated, CXCR3(+) and CCR5(+) cells were observed in these lesions. Cells positive for STAT1 and chemokine CXCL9, interferon- (IFN)-induced gene products, and pSTAT1 were also observed in the same lesions. Double immunofluorescence staining demonstrated that the cells that were positive for CD163 were also positive for STAT1.

Conclusions: CD163(+) TAMs in oral premalignant lesions coexpress CD163 and STAT1, suggesting that the TAMs in oral premalignant lesions possess an M1 phenotype in a Th1-dominated micromilieu.

No MeSH data available.


Related in: MedlinePlus

Colocalization of CD163+ TAMS with STAT1 in oral leukoplakia. Double-labeled fluorescent immunostaining for CD163 (green) and STAT1 or pSTAT1 (red) in oral leukoplakia with moderate dysplasia. The cells double-stained for both the anti-CD163 and anti-STAT1 antibodies are shown (a, yellow). These cells colocalized to the subepithelial lesion (original magnification: ×200). Scale bar = 100 μm. b Correlation between infiltrated STAT1+ and CD163+ cells in oral leukoplakia. Statistically significant differences were determined using Spearman’s rank correlation coefficient analysis (p = 0.0034). c The cells double-stained for both the anti-CD163 and anti-pSTAT1 antibodies are shown (arrow heads)
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Fig5: Colocalization of CD163+ TAMS with STAT1 in oral leukoplakia. Double-labeled fluorescent immunostaining for CD163 (green) and STAT1 or pSTAT1 (red) in oral leukoplakia with moderate dysplasia. The cells double-stained for both the anti-CD163 and anti-STAT1 antibodies are shown (a, yellow). These cells colocalized to the subepithelial lesion (original magnification: ×200). Scale bar = 100 μm. b Correlation between infiltrated STAT1+ and CD163+ cells in oral leukoplakia. Statistically significant differences were determined using Spearman’s rank correlation coefficient analysis (p = 0.0034). c The cells double-stained for both the anti-CD163 and anti-pSTAT1 antibodies are shown (arrow heads)

Mentions: To further characterize the CD163+ macrophages in leukoplakia, we examined the coexpression of CD163 and STAT1 or pSTAT1 using double-labeling immunofluorescence (Fig. 5). CD163+ macrophages were distributed in the subepithelial lesion, and the majority of CD163+ cells located in the papillary dermis colocalized with STAT1 (Fig. 5a). The percentages of CD163+ macrophages and STAT1+ cells were positively correlated (p = 0.0034; Fig. 5b). Although the percentages of single-stained cells for CD163 and STAT1 were 16.4 % and 32.1 %, respectively, the percentage of double-stained cells was 51.5 % (n = 4). The CD163+ cells also coexpressed pSTAT1 (Fig. 5c). These results indicate that CD163+ macrophages in oral leukoplakia coexpress active STAT1 and suggest that the CD163+ macrophages possess an M1 phenotype in a Th1-dominated microenvironment.Fig. 5


Tumor-associated macrophages in oral premalignant lesions coexpress CD163 and STAT1 in a Th1-dominated microenvironment.

Mori K, Haraguchi S, Hiori M, Shimada J, Ohmori Y - BMC Cancer (2015)

Colocalization of CD163+ TAMS with STAT1 in oral leukoplakia. Double-labeled fluorescent immunostaining for CD163 (green) and STAT1 or pSTAT1 (red) in oral leukoplakia with moderate dysplasia. The cells double-stained for both the anti-CD163 and anti-STAT1 antibodies are shown (a, yellow). These cells colocalized to the subepithelial lesion (original magnification: ×200). Scale bar = 100 μm. b Correlation between infiltrated STAT1+ and CD163+ cells in oral leukoplakia. Statistically significant differences were determined using Spearman’s rank correlation coefficient analysis (p = 0.0034). c The cells double-stained for both the anti-CD163 and anti-pSTAT1 antibodies are shown (arrow heads)
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4525742&req=5

Fig5: Colocalization of CD163+ TAMS with STAT1 in oral leukoplakia. Double-labeled fluorescent immunostaining for CD163 (green) and STAT1 or pSTAT1 (red) in oral leukoplakia with moderate dysplasia. The cells double-stained for both the anti-CD163 and anti-STAT1 antibodies are shown (a, yellow). These cells colocalized to the subepithelial lesion (original magnification: ×200). Scale bar = 100 μm. b Correlation between infiltrated STAT1+ and CD163+ cells in oral leukoplakia. Statistically significant differences were determined using Spearman’s rank correlation coefficient analysis (p = 0.0034). c The cells double-stained for both the anti-CD163 and anti-pSTAT1 antibodies are shown (arrow heads)
Mentions: To further characterize the CD163+ macrophages in leukoplakia, we examined the coexpression of CD163 and STAT1 or pSTAT1 using double-labeling immunofluorescence (Fig. 5). CD163+ macrophages were distributed in the subepithelial lesion, and the majority of CD163+ cells located in the papillary dermis colocalized with STAT1 (Fig. 5a). The percentages of CD163+ macrophages and STAT1+ cells were positively correlated (p = 0.0034; Fig. 5b). Although the percentages of single-stained cells for CD163 and STAT1 were 16.4 % and 32.1 %, respectively, the percentage of double-stained cells was 51.5 % (n = 4). The CD163+ cells also coexpressed pSTAT1 (Fig. 5c). These results indicate that CD163+ macrophages in oral leukoplakia coexpress active STAT1 and suggest that the CD163+ macrophages possess an M1 phenotype in a Th1-dominated microenvironment.Fig. 5

Bottom Line: Although CCR4(+) cells rarely infiltrated, CXCR3(+) and CCR5(+) cells were observed in these lesions.Cells positive for STAT1 and chemokine CXCL9, interferon- (IFN)-induced gene products, and pSTAT1 were also observed in the same lesions.Double immunofluorescence staining demonstrated that the cells that were positive for CD163 were also positive for STAT1.

View Article: PubMed Central - PubMed

Affiliation: Division of Oral and Maxillofacial Surgery, Department of Diagnosis and Therapeutics Sciences, Meikai University of School of Dentistry, 1-1 Keyakidai, Sakado, Saitama, 350-0283, Japan. kazu-mori@dent.meikai.ac.jp.

ABSTRACT

Background: Tumor-associated macrophages (TAMs) are implicated in the growth, invasion and metastasis of various solid tumors. However, the phenotype of TAMs in premalignant lesions of solid tumors has not been clarified. In the present study, we identify the phenotype of TAMs in leukoplakia, an oral premalignant lesion, by immunohistochemical analysis and investigate the involvement of infiltrated T cells that participate in the polarization of TAMs.

Methods: The subjects included 30 patients with oral leukoplakia and 10 individuals with normal mucosa. Hematoxylin and eosin slides were examined for the histological grades, and immunohistochemical analysis was carried out using antibodies against CD68 (pan-MΦ), CD80 (M1 MΦ), CD163 (M2 MΦ), CD4 (helper T cells: Th), CD8 (cytotoxic T cells), CXCR3, CCR5 (Th1), CCR4 (Th2), signal transducer and activator of transcription (STAT1), phosphorylated STAT1 (pSTAT1) and chemokine CXCL9. The differences in the numbers of positively stained cells among the different histological grades were tested for statistical significance using the Kruskal-Wallis test. Correlations between different types of immune cells were determined using Spearman's rank analysis.

Results: An increase in the rate of CD163(+) TAM infiltration was observed in mild and moderate epithelial dysplasia, which positively correlated with the rate of intraepithelial CD4(+) Th cell infiltration. Although CCR4(+) cells rarely infiltrated, CXCR3(+) and CCR5(+) cells were observed in these lesions. Cells positive for STAT1 and chemokine CXCL9, interferon- (IFN)-induced gene products, and pSTAT1 were also observed in the same lesions. Double immunofluorescence staining demonstrated that the cells that were positive for CD163 were also positive for STAT1.

Conclusions: CD163(+) TAMs in oral premalignant lesions coexpress CD163 and STAT1, suggesting that the TAMs in oral premalignant lesions possess an M1 phenotype in a Th1-dominated micromilieu.

No MeSH data available.


Related in: MedlinePlus