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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-CXCR3 and anti-CCR5 antibodies. Immunoreactivity against anti-CXCR3 (a, b) and anti-CCR5 (c, d) antibodies for moderate grades of oral leukoplakia (original magnification: A, C: ×100; B, D: ×400). CXCR3+ and CCR5+ Th1 cells were mainly distributed in the subepithelial lesion. Scale bar = 100 μm (a), 300 μm (c), and 30 μm (b, d)
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Fig3: Immunohistochemical staining of oral leukoplakia with the anti-CXCR3 and anti-CCR5 antibodies. Immunoreactivity against anti-CXCR3 (a, b) and anti-CCR5 (c, d) antibodies for moderate grades of oral leukoplakia (original magnification: A, C: ×100; B, D: ×400). CXCR3+ and CCR5+ Th1 cells were mainly distributed in the subepithelial lesion. Scale bar = 100 μm (a), 300 μm (c), and 30 μm (b, d)

Mentions: Th1-derived IFN reportedly induces classically activated M1 macrophages, whereas Th2-derived IL-4 and IL-13 induce alternatively activated M2 macrophages [12, 19]. To further analyze the subset of infiltrated T cells that affect the phenotype of TAMs, we immunohistochemically examined the infiltrated CD4+ T cells using antibodies to chemokine receptor CXCR3 and CCR5, markers for Th1 cells, and antibodies to CCR4, a marker for Th2 cells [34]. Although CCR4+ T cells were rare in normal mucosa and in leukoplakia lesions (data not shown), CXCR3+ (Fig. 3a, b) and CCR5+ (Fig. 3c, d) T cells abundantly infiltrated the subepithelial stroma of leukoplakia. These results indicate that CD4+ Th1 cells are the predominant subset of T cells that infiltrate leukoplakia.Fig. 3


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-CXCR3 and anti-CCR5 antibodies. Immunoreactivity against anti-CXCR3 (a, b) and anti-CCR5 (c, d) antibodies for moderate grades of oral leukoplakia (original magnification: A, C: ×100; B, D: ×400). CXCR3+ and CCR5+ Th1 cells were mainly distributed in the subepithelial lesion. Scale bar = 100 μm (a), 300 μm (c), and 30 μm (b, d)
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Related In: Results  -  Collection

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Fig3: Immunohistochemical staining of oral leukoplakia with the anti-CXCR3 and anti-CCR5 antibodies. Immunoreactivity against anti-CXCR3 (a, b) and anti-CCR5 (c, d) antibodies for moderate grades of oral leukoplakia (original magnification: A, C: ×100; B, D: ×400). CXCR3+ and CCR5+ Th1 cells were mainly distributed in the subepithelial lesion. Scale bar = 100 μm (a), 300 μm (c), and 30 μm (b, d)
Mentions: Th1-derived IFN reportedly induces classically activated M1 macrophages, whereas Th2-derived IL-4 and IL-13 induce alternatively activated M2 macrophages [12, 19]. To further analyze the subset of infiltrated T cells that affect the phenotype of TAMs, we immunohistochemically examined the infiltrated CD4+ T cells using antibodies to chemokine receptor CXCR3 and CCR5, markers for Th1 cells, and antibodies to CCR4, a marker for Th2 cells [34]. Although CCR4+ T cells were rare in normal mucosa and in leukoplakia lesions (data not shown), CXCR3+ (Fig. 3a, b) and CCR5+ (Fig. 3c, d) T cells abundantly infiltrated the subepithelial stroma of leukoplakia. These results indicate that CD4+ Th1 cells are the predominant subset of T cells that infiltrate leukoplakia.Fig. 3

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