Tumor-infiltrating CD4+ and CD8+ lymphocytes and macrophages are associated with prognostic factors in triple-negative canine mammary complex type carcinoma.

This study aimed to evaluate the association of CD3+, CD4+, and CD8+ T cells and tumor-infiltrating macrophages (TIMs) with the clinical parameters of female dogs harboring mammary gland tumors. Thirty female dogs affected with mammary carcinomas were used, and all tumors were histologically classified as complex carcinoma and were triple-negative phenotype determined by immunohistochemistry. Freshly frozen sections were used to determine CD3+, CD4+ and CD8+ T cells by immunohistochemistry, and TIMs were determined by immunofluorescence assays. Ten out of the 30 dogs showed lymph node metastasis at diagnosis. Fifteen dogs had a tumor of grade I (15/30), nine (9/30) had a tumor of grade II and six (6/30) had a tumor of grade III. The mean overall survival was 680.5 days (± 200.4). Dogs with sentinel lymph node positivity (10/30) (P = .0035) and dogs that developed metastasis (P = .0001) showed a shorter survival time. In addition, dogs with a high level of inflammatory infiltrate in tumor tissues presented a shorter survival time (P = .0001) than that of other dogs. Dogs with tumors containing higher numbers of CD3+ T cells (P = .001), CD4+ T cells (P = .001), or TIM cells (P < .0001) showed a shorter survival time than that of other dogs. Our results suggested that characteristics of immune cell infiltrates, including CD3+ T cells, CD4+ T cells, and TIMs, can be used as potential prognostic indicators for predicting clinical outcomes in dogs with mammary gland tumors, particularly tumors with a complex histological subtype and triple-negative phenotype.

Tumor-infiltrating lymphocytes in Breast Cancer and implications for clinical practice.

Breast Cancer (BC) can be classified using pathologic features, such as grade and tumor size. It can be categorized based on the gene expression profile, which identifies the distinct molecular subtype. More recently, stromal tissue has been recognized as an important modulator of tumor cell growth, pathogenesis, and progression. Immune cells could drive important clinical characteristics that affect BC outcomes. Subgroups of patients who have tumor-infiltrating lymphocytes in the stroma may have better response to chemotherapy and favorable long-term prognosis. Accumulating evidence shows that the immune system plays a crucial role in the outcomes of some BC subgroups, especially more aggressive, proliferative ones such as triple-negative and HER2-positive BC. This review article will present data on the role of lymphocyte infiltration in BC prognosis and response to therapy. This review will also introduce the reader to the challenges of applying this promising prognostic and predictive biomarker in clinical practice.

Indicadores de riesgo de recidiva local en el tratamiento conservador del cáncer de mama / Risk indicators of local relapse in the conservative treatment of the breast cancer

Fueron evaluados los posibles factores de riesgo para recidiva mamaria en pacientes con estadio I y II de adenocarcinoma ductal invasor, sometidas a tratamiento conservador. Luego de un seguimiento promedio de 60 meses, se registraron 20 recidivas en 211 pacientes con tratamiento conservador. Sobre un total de 152 pacientes se realizó un análisis estadístico, uni y multivariado, para hallar los factores de significación respecto al riesgo de recidiva. Igual metodología de análisis fue implementada en pacientes pre y postmenopáusicas por separado. Se identificaron dos tipos de factores, aquellos asociados con la agresividad tumoral y los relacionados con la enfermedad residual. Tanto para pacientes premenopáusicas como postmenopáusicas, el factor de riesgo más significativo del primer tipo, fue la invasión vascular. Entre los factores asociados a enfermedad residual. Tanto para pacientes premenopáusicas como postmenopáusicas, el factor de riesgo más significativo del primer tipo, fue la invasión vascular. Entre los factores asociados a enfermedad residual, resultó significativo la presencia de componente intraductal extenso. Los bordes quirúrgicos comprometidos resultaron estadísticamente significativos sólo en pacientes postmenopáusicas.

Indicadores de riesgo de recidiva local en el tratamiento conservador del cáncer de mama / Risk indicators of local relapse in the conservative treatment of the breast cancer

Fueron evaluados los posibles factores de riesgo para recidiva mamaria en pacientes con estadio I y II de adenocarcinoma ductal invasor, sometidas a tratamiento conservador. Luego de un seguimiento promedio de 60 meses, se registraron 20 recidivas en 211 pacientes con tratamiento conservador. Sobre un total de 152 pacientes se realizó un análisis estadístico, uni y multivariado, para hallar los factores de significación respecto al riesgo de recidiva. Igual metodología de análisis fue implementada en pacientes pre y postmenopáusicas por separado. Se identificaron dos tipos de factores, aquellos asociados con la agresividad tumoral y los relacionados con la enfermedad residual. Tanto para pacientes premenopáusicas como postmenopáusicas, el factor de riesgo más significativo del primer tipo, fue la invasión vascular. Entre los factores asociados a enfermedad residual. Tanto para pacientes premenopáusicas como postmenopáusicas, el factor de riesgo más significativo del primer tipo, fue la invasión vascular. Entre los factores asociados a enfermedad residual, resultó significativo la presencia de componente intraductal extenso. Los bordes quirúrgicos comprometidos resultaron estadísticamente significativos sólo en pacientes postmenopáusicas. (AU)

Usage of T-cell receptor V beta chain genes in fresh and cultured tumor-infiltrating lymphocytes from human melanoma.

Tumor-infiltrating lymphocytes (TIL) freshly obtained from human malignant melanomas as well as the same TIL grown in the presence of interleukin 2 (IL2) were studied for gene expression of the T-cell receptor (TCR) variable beta regions (V beta). To perform the TCR-V beta analysis, total RNA was isolated from TIL and reverse-transcribed into cDNA, which was then amplified by PCR using 22 different 5' primers specifically recognizing the sequences of 20 V beta gene families and a 3' primer annealing to the constant region of the beta chain. The TCR-alpha constant region (C alpha) gene was co-amplified as a standard for the calculation of the percentage of each TCR-V beta gene expressed. The frequency of individual V beta regions expressed on TIL was computed from the ratio of cpm V beta to cpm C alpha for each V beta region in relation to the total of all 22 ratios. With fresh TIL obtained from 8 different melanomas, oligoclonal distribution of V beta genes expressed on TIL was observed, in comparison with a broader and unrestricted distribution seen with peripheral-blood T cells of 8 normal individuals. The oligoclonal patterns of V beta-gene expression in fresh melanoma TIL were distinct in every tumor. Several of the V beta-genes usually expressed in normal PBL were not expressed in fresh TIL in melanoma TIL cultured in the presence of IL2 and IL4 and in the absence of autologous tumor (AuTu) or antigen-presenting cells for 23 to 65 days, selection of T-cell lines expressing a restricted number of V beta genes occurred. Although in 4/5 TIL cultures this selection involved the V beta 7 gene, no relationship could be established between V beta gene expression in fresh TIL and that in T-cell lines outgrowing in long-term cultures. Selection in culture of CD3+CD8+ T-cell lines with V beta-gene expression restricted to 1 or 2 V beta families did not correlate with the presence or level of AuTu cytotoxicity mediated by these T cells. The results indicate that in TIL cultures random selection of T-cell lines with reactivity not relevant to AuTu may account for poor expression or loss of AuTu cytotoxicity by most TIL cultured long-term in the presence of cytokines and in the absence of specific antigenic stimulation.

Clonal analysis of tumor-infiltrating lymphocytes from human primary and metastatic liver tumors.

Phenotypic and functional characteristics of tumor-infiltrating lymphocytes (TIL) obtained from human primary and metastatic liver tumors were studied. Lymphocytes isolated from 18 tumors and autologous (A) peripheral blood (6 cases) were phenotyped by 2-color flow cytometry and cloned in a limiting dilution system, which allows virtually all normal T lymphocytes to proliferate; 70-80% of fresh TIL were T cells (i.e., CD3+), and the ratio of CD4+/CD8+ cells was 1.2 in both primary and metastatic liver tumors. TIL contained significantly more CD56+ (NKHI+) cells, half of which were CD3+CD56+, CD3+CD25+ cells and CD3+HLA-DR+ cells, than A-PBL. The frequencies of proliferating T-cell precursors (PTL-p) and cytolytic T-lymphocyte precursors (CTL-p) reactive with K562, allogeneic tumor cells and autologous tumor cells, were determined. Mean PTL-p frequencies for TIL from hepatocellular carcinomas, cholangiocarcinomas and metastatic liver tumors were 0.52 (0.22-0.83), 0.10 (0.05-0.16) and 0.16 (0.01-0.30), respectively. The frequency of CTL-p with natural-killer-like activity was lower in TIL than in A-PBL. The frequency of CTL-p for autologous tumor cells in fresh TIL isolated from primary liver tumors was 0.02-0.13 and 12/81 clones were reactive against autologous tumor. In contrast, only 1/66 TIL clones obtained from colon carcinomas metastatic to liver showed autotumor reactivity. No clones reactive with autologous tumor were obtained from peripheral blood of patients with liver cancer. These data indicate that substantial differences in anti-tumor functions of TIL between primary and metastatic liver tumors exist, which can be detected at a clonal level.