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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

Immunohistochemical staining of oral leukoplakia with the anti-STAT1, anti-pSTAT1 and anti-CXCL9/Mig antibodies Immunoreactivity against anti-STAT1 (a, b), anti-tyrosine (Try701)-phosphorylated- STAT1 (pSTAT1) (d, e) and anti-CXCL9/Mig (f, g) antibodies for moderate grades of oral leukoplakia (original magnification: A, D, F: ×100; B, E, G: ×400). STAT1+ and CXCL9+ cells were mainly distributed in the subepithelial lesion. Scale bar = 300 μm (a, d), 100 μm (f), 50 μm (e, g) and 30 μm (b). Correlation between infiltrated CXCR3+ and STAT1+ cells in leukoplakia (c). Statistically significant differences were determined using Spearman’s rank correlation coefficient analysis (p = 0.0465)
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Fig4: Immunohistochemical staining of oral leukoplakia with the anti-STAT1, anti-pSTAT1 and anti-CXCL9/Mig antibodies Immunoreactivity against anti-STAT1 (a, b), anti-tyrosine (Try701)-phosphorylated- STAT1 (pSTAT1) (d, e) and anti-CXCL9/Mig (f, g) antibodies for moderate grades of oral leukoplakia (original magnification: A, D, F: ×100; B, E, G: ×400). STAT1+ and CXCL9+ cells were mainly distributed in the subepithelial lesion. Scale bar = 300 μm (a, d), 100 μm (f), 50 μm (e, g) and 30 μm (b). Correlation between infiltrated CXCR3+ and STAT1+ cells in leukoplakia (c). Statistically significant differences were determined using Spearman’s rank correlation coefficient analysis (p = 0.0465)

Mentions: Because Th1 cells produce IFN, which induces M1 macrophages, we next assessed whether the IFN-inducible gene products STAT1 [35] and CXCL9/Mig, a chemokine for Th1 [36], were expressed in leukoplakia (Fig. 4). STAT1+ cells were widely distributed in the subepithelial lesions of leukoplakia. The percentages of CXCR3+ cells positively correlated with the percentages of STAT1+ cells (p = 0.0465; Fig. 4c). Tyrosine-phosphorylated STAT1 (pSTAT1), an active form of STAT1, was also detected in the lesions (Fig. 4d, e), though the frequency of pSTAT1-positive cells was lower than that of STAT1-positive cells. Cells positive for the IFN-inducible chemokine CXCL9 were also observed in the subepithelial lesion of leukoplakia (Fig. 4f, g). Taken together, these results indicate that the leukoplakia lesions form a Th1-dominated microenvironment and suggest that Th1-derived IFN affects the infiltrated macrophages to polarize the M1 phenotype.Fig. 4


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)

Immunohistochemical staining of oral leukoplakia with the anti-STAT1, anti-pSTAT1 and anti-CXCL9/Mig antibodies Immunoreactivity against anti-STAT1 (a, b), anti-tyrosine (Try701)-phosphorylated- STAT1 (pSTAT1) (d, e) and anti-CXCL9/Mig (f, g) antibodies for moderate grades of oral leukoplakia (original magnification: A, D, F: ×100; B, E, G: ×400). STAT1+ and CXCL9+ cells were mainly distributed in the subepithelial lesion. Scale bar = 300 μm (a, d), 100 μm (f), 50 μm (e, g) and 30 μm (b). Correlation between infiltrated CXCR3+ and STAT1+ cells in leukoplakia (c). Statistically significant differences were determined using Spearman’s rank correlation coefficient analysis (p = 0.0465)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4525742&req=5

Fig4: Immunohistochemical staining of oral leukoplakia with the anti-STAT1, anti-pSTAT1 and anti-CXCL9/Mig antibodies Immunoreactivity against anti-STAT1 (a, b), anti-tyrosine (Try701)-phosphorylated- STAT1 (pSTAT1) (d, e) and anti-CXCL9/Mig (f, g) antibodies for moderate grades of oral leukoplakia (original magnification: A, D, F: ×100; B, E, G: ×400). STAT1+ and CXCL9+ cells were mainly distributed in the subepithelial lesion. Scale bar = 300 μm (a, d), 100 μm (f), 50 μm (e, g) and 30 μm (b). Correlation between infiltrated CXCR3+ and STAT1+ cells in leukoplakia (c). Statistically significant differences were determined using Spearman’s rank correlation coefficient analysis (p = 0.0465)
Mentions: Because Th1 cells produce IFN, which induces M1 macrophages, we next assessed whether the IFN-inducible gene products STAT1 [35] and CXCL9/Mig, a chemokine for Th1 [36], were expressed in leukoplakia (Fig. 4). STAT1+ cells were widely distributed in the subepithelial lesions of leukoplakia. The percentages of CXCR3+ cells positively correlated with the percentages of STAT1+ cells (p = 0.0465; Fig. 4c). Tyrosine-phosphorylated STAT1 (pSTAT1), an active form of STAT1, was also detected in the lesions (Fig. 4d, e), though the frequency of pSTAT1-positive cells was lower than that of STAT1-positive cells. Cells positive for the IFN-inducible chemokine CXCL9 were also observed in the subepithelial lesion of leukoplakia (Fig. 4f, g). Taken together, these results indicate that the leukoplakia lesions form a Th1-dominated microenvironment and suggest that Th1-derived IFN affects the infiltrated macrophages to polarize the M1 phenotype.Fig. 4

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