RESUMEN
OBJECTIVE: To evaluate and characterize macrophage populations (M1/M2) in the tumor microenvironment of oral cavity squamous cell carcinoma (OCSCC). The relationship between macrophages and clinicopathological factors, such as survival data, lymph node metastasis, tumoral proliferation, and WHO histological grading are also analyzed. MATERIALS AND METHODS: The samples consisted of surgically excised specimens from patients with non-metastatic and metastatic OCSCC and normal oral mucosa (control). Immunohistochemistry, flow cytometry, and qRT-PCR were used to evaluate macrophage populations and the expression of pro- (IL-12, IL-23, and INF-γ) and anti-inflammatory (IL-10 and TGF-ß) cytokines. The level required for statistical significance was defined as p<0.05. RESULTS: The data showed a predominance of M2 phenotype (high percentage of IL-10(+)TGF-ß(+)) macrophages in the tumor microenvironment of OCSCC. A higher percentage of macrophages expressing TGF-ß was seen in the OCSCC group when compared with healthy individuals. The assessment of mRNA expression also presented a greater expression of anti-inflammatory cytokines TGFß and IL10 in OCSCC when compared with the control group. The percentage of macrophages, demonstrated by immunohistochemistry, was significantly higher in the metastatic OCSCC group than in the non-metastatic and control groups. The log-rank test also showed that the mean survival time for patients with high levels of macrophages was less (44 months) when compared with patients with a low percentage of such cells (93 months). CONCLUSION: A predominance of the M2 phenotype in the tumor microenvironment of OCSCC could contribute to local immunosuppression, via TGF-ß production, and consequently greater lymph node involvement and reduced patient survival time.
Asunto(s)
Carcinoma de Células Escamosas/inmunología , Citocinas/análisis , Mediadores de Inflamación/análisis , Macrófagos/inmunología , Neoplasias de la Boca/inmunología , Adulto , Anciano , Anciano de 80 o más Años , Antígenos CD11/análisis , Carcinoma de Células Escamosas/patología , Carcinoma de Células Escamosas/secundario , Recuento de Células , Proliferación Celular , Femenino , Estudios de Seguimiento , Humanos , Tolerancia Inmunológica/inmunología , Interferón gamma/análisis , Interleucina-10/análisis , Interleucina-12/análisis , Interleucina-23/análisis , Metástasis Linfática/inmunología , Metástasis Linfática/patología , Macrófagos/clasificación , Macrófagos/patología , Masculino , Persona de Mediana Edad , Neoplasias de la Boca/patología , Clasificación del Tumor , Invasividad Neoplásica , Estudios Retrospectivos , Tasa de Supervivencia , Factor de Crecimiento Transformador beta/análisis , Microambiente Tumoral/inmunologíaRESUMEN
Cigarette smoking produces peripheral airway inflammation in all smokers, and chronic airways obstruction in approximately 20% of heavy smokers. The present study was designed to test the hypothesis that airways obstruction is related to changes in the expression of adhesion molecules involved in the recruitment of cells to sites of inflammation in the lung. Freshly resected lungs from heavy smokers with airways obstruction (n = 10) and from heavy smokers with normal lung function (n = 10) were collected in the operating room, inflated with optimal cutting temperature (OCT) medium and frozen over liquid nitrogen. Six micrometres thick cryostat sections cut from random samples of this tissue were stained, using immunohistochemistry, with monoclonal antibodies to the adhesion molecules on leucocytes: L-selectin, very late activation antigen-4 (VLA-4), CD11a/CD18, CD11b/CD18, CD11c/CD18; and on endothelial and epithelial surfaces: E-selectin, P-selectin, vascular cell adhesion molecule (VCAM-1), intercellular adhesion molecule (ICAM)-1 and ICAM-2 using the alkaline phosphatase anti-alkaline phosphatase (APAAP) technique. The slides were coded and the expression of each molecule scored by three observers using a semiquantitative grading system. Two inducible adhesion molecules, E-selectin on endothelium and CD11b on leucocytes, were also evaluated using quantitative morphometric analysis. The results showed a distribution of adhesion molecules that was consistent with the inflammatory response in the airways and parenchyma of all subjects but failed to show any differences between those with or without airways obstruction. We conclude that development of airways obstruction in heavy smokers cannot be explained by differences in the expression of adhesion molecules known to be involved in the control of cell traffic in the lung.
Asunto(s)
Moléculas de Adhesión Celular/genética , Regulación de la Expresión Génica , Enfermedades Pulmonares Obstructivas/metabolismo , Fumar/metabolismo , Anciano , Anticuerpos Monoclonales , Antígenos CD/análisis , Antígenos CD/genética , Antígenos CD11/análisis , Antígenos CD11/genética , Antígenos CD18/análisis , Antígenos CD18/genética , Moléculas de Adhesión Celular/análisis , Selectina E/análisis , Selectina E/genética , Endotelio/metabolismo , Endotelio/patología , Epitelio/metabolismo , Epitelio/patología , Femenino , Humanos , Inmunohistoquímica , Integrina alfa4beta1 , Integrina beta1/análisis , Integrina beta1/genética , Integrinas/análisis , Integrinas/genética , Molécula 1 de Adhesión Intercelular/análisis , Molécula 1 de Adhesión Intercelular/genética , Selectina L/análisis , Selectina L/genética , Leucocitos/metabolismo , Leucocitos/patología , Pulmón/metabolismo , Pulmón/patología , Enfermedades Pulmonares Obstructivas/genética , Enfermedades Pulmonares Obstructivas/patología , Masculino , Persona de Mediana Edad , Selectina-P/análisis , Selectina-P/genética , Neumonía/genética , Neumonía/metabolismo , Neumonía/patología , Receptores Mensajeros de Linfocitos/análisis , Receptores Mensajeros de Linfocitos/genética , Receptores de Antígeno muy Tardío/análisis , Receptores de Antígeno muy Tardío/genética , Humo , Fumar/genética , Fumar/patología , Molécula 1 de Adhesión Celular Vascular/análisis , Molécula 1 de Adhesión Celular Vascular/genéticaRESUMEN
Because adhesion properties of leukocytes are important for the influx and localization of leukocytes in sites of inflammation, we studied, the expression of intercellular adhesion molecule 1 (ICAM-1), lymphocyte-function-associated antigen 1 (LFA-1), vascular cell adhesion molecule- 1 (VCAM-1) and CD 11b in 14 kidney biopsies of PSGN patients, arbitrarily divided into early biopsies (less than 15 days after onset of PSGN) and late biopsies (17-90 days). In PSGN, intraglomerular ICAM-1 expression was increased in early biopsies (score 3.1 +/- SEM 0.2; P < 0.005) and decreased with time; in late biopsies the score (2.0 +/- 0.2) was similar to that of normal kidney (1.3 +/- 0.3). In the interstitium ICAM-1 was increased (early PSGN = 836 +/- 56 positive cells/mm2, late = 552 +/- 60.0; versus normal = 364 +/- 12.4; P < 0.05). LFA-1 expressing cells in glomeruli were also increased in early biopsies (10.0 +/- 2.1 positive cells per glomerular cross-section (gcs), versus normal 2.9 +/- 1.4; P < 0.05). In the interstitium, LFA-1 positive cells were increased (early PSGN = 221 +/- 79.6 cells/mm2, late PSGN = 134.5 +/- 45.1, normal = 21 +/- 8.7; P < 0.05). VCAM-1 in glomeruli and interstitium was not increased in PSGN. Our studies demonstrate increased expression of adhesion molecules ICAM-1 and LFA-1 in the kidney of PSGN patients, and these findings were more pronounced in early biopsies; adhesion molecules are probably involved in the inflammatory infiltration of this disease.