Tumoral inflammatory infiltrate does not predict metastases in thin primary cutaneous melanomas

Abstract Background In cutaneous melanomas in general, tumor inflammatory infiltrate (TII) can protect against distant metastases, but there is no consensus when only thin primary cutaneous melanomas (TPCM) are considered. Objective To investigate the presence of TII in TPCM and the relationship between TII and the occurrence of metastases. Methods Case-control study including 50 patients with TPCM, 22 metastatic (MC group) and 28 non-metastatic (NMC group). The presence of TII was evaluated and, if present, qualified as mild, moderate or marked. Results The mean age was 50.7 years in the MC and 56.2 years in the NMC group (p = 0.234), and the male sex predominated in the MC group (63.6%). The average Breslow thickness was higher in the MC when compared to that observed in the NMC (respectively 0.8 vs. 0.6 mm, p = 0.012). The presence of ulceration occurred in 22.7% of the MC and 17.9% of the NMC (p = 0.732). TII was present in all 50 TPCM, being marked or moderate in 67.9% of the NMC and 54.5% in the MC group (p = 0.503). In the multivariate analysis, the presence of moderate and marked TII had an Odds Ratio (OR) of 0.57 (95% Confidence Interval [CI]: 0.18‒1.8) and adjusted OR of 0.68 (95% CI 0.13‒3.99). Study limitations Small sample size. Conclusions TII was present in all TPCM (with and without metastases), and it was not possible to demonstrate a protective effect of TII against the appearance of metastases.

Research advances in tumor-infiltrating lymphocytes in treatment of biliary tract cancer / 临床肝胆病杂志

Tumor-infiltrating lymphocytes (TILs) are a heterogeneous subset of lymphocytes, mainly T cells, present in tumor parenchyma and stroma. After being digested and isolated from tumor tissue and then cultured in vitro for activation and multiplication, it can be infused back into the patient's body to kill tumor cells. TILs have the advantages of high diversity of TCR, excellent ability to infiltrate into tumor sites, and low toxicity and are considered promising for the treatment of malignant solid tumors. At present, TIL therapy has been tested as a second-line treatment in a variety of solid tumors and has achieved preliminary results. Although there is still no clinical cohort report on the application of TILs in biliary tract cancer (BTC), recent clinical reports on multiple cancers have provided information on the efficacy of TIL therapy in a small number of BTC patients, which preliminarily confirmed the safety and efficacy of TIL therapy. However, since BTC is generally considered an immunologically repulsive tumor in which most effector T cells are sequestered at the tumor edge, the antitumor effect of TILs in BTC remains difficult to predict. Combination therapy with different anti-tumor methods and the development of new techniques to modify cells to enhance the anti-tumor ability of TILs are possible directions for breakthrough in the future.

Clinical significance of CD8 positive tumor-infiltrating lymphocytes density and programmed-death receptor ligand 1 expression in rectal cancer after neoadjuvant chemoradiotherapy / 肿瘤研究与临床

Objective:To investigate the correlation of CD8 positive tumor-infiltrating lymphocytes (CD8 + TIL) density and programmed-death receptor ligand 1 (PD-L1) expression in rectal cancer with clinicopathological characteristics and prognosis of patients after neoadjuvant chemoradiotherapy. Methods:The clinicopathological data of 166 patients with locally advanced rectal cancer (LARC) who received neoadjuvant therapy before surgery in the Beijing Chao-Yang Hospital, Capital Medical University from January 2015 to December 2018 were retrospectively analyzed. CD8 + TIL density and PD-L1 expression were detected by using immunohistochemistry. The correlation of CD8 + TIL density and PD-L1 expression with clinicopathological characteristics of patients after neoadjuvant chemoradiotherapy was analyzed. Kaplan-Meier method was used to analyze the disease-free survival (DFS) and Cox regression risk model was used to make univariate and multivariate analysis of the influencing factors for DFS. Results:Among 166 LARC patients, 81 cases (48.8%) had high density of CD8 + TIL, 85 cases (51.2%) had low density of CD8 + TIL; 63 cases (38.0%) had PD-L1 expression, and 103 cases (62.0%) had non-expression of CD8 + TIL. The expression rate of PD-L1 in CD8 + TIL high density group was higher than that in CD8 + TIL low density group [50.6% (41/81) vs. 25.9%(22/85), χ2 = 10.78, P < 0.001]. According to the density of CD8 + TIL and PD-L1 expression, immunophenotype was divided among 4 groups; the 3-year DFS rate of the CD8 + TIL high density /PD-L1 expression group was 87.1%, which was higher than that of the other groups (CD8 + TIL low density /PD-L1 expression group was 72.8%, CD8 + TIL high density /PD-L1 non-expression group was 67.0%, CD8 + TIL low density /PD-L1 non-expression group was 64.3%), and the difference was statistically significant ( P < 0.05). Univariate analysis showed that tumor differentiation degree, TNM stage, CD8 + TIL density, PD-L1 expression and CD8 + TIL density /PD-L1 expression were correlated with the DFS of patients (all P < 0.05). Multivariate analysis results showed that CD8 + TIL high density /PD-L1 expression was an independent protective factor for DFS ( HR = 0.049, 95% CI 0.005-0.497, P = 0.011), while TNM stage 3 was an independent risk factor for DFS ( HR = 2.752,95% CI 1.300-5.825, P = 0.008). Conclusions:In LARC after neoadjuvant therapy, CD8 + TIL density is positively correlated with the expression of PD-L1, and the high density of CD8 + TIL/PD-L1 expression is an independent influencing factor for good prognosis, suggesting that these patients may benefit from the immunotherapy.

SPECT/CT imaging of tumor-infiltrating CD8 + T cell to predict the efficacy of anti-PD-1 immunotherapy in mice / 中华核医学与分子影像杂志

Objective:To prepare 99Tc m-hydrazinonicotinamide(HYNIC)-αCD8/Fab ( 99Tc m-αCD8/Fab), and explore the predictive value of 99Tc m-αCD8/Fab SPECT/CT imaging for the efficacy of anti-programmed death-1 (PD-1) immunotherapy. Methods:The αCD8/Fab was modified with HYNIC- N-hydroxysuccinimide (NHS) and IRDye800-NHS to form HYNIC-αCD8/Fab and IRDye800-αCD8/Fab (Dye-αCD8/Fab), respectively. 99Tc m-αCD8/Fab was prepared in sodium bicarbonate buffer (pH=8.5), with SnCl 2 being used as the reducing agent. The labeling yield and radiochemical purity of 99Tc m-αCD8/Fab and its stability in PBS and fetal bovine serum (FBS) were tested in vitro. The mouse spleen and human peripheral blood lymphocytes were isolated for cell-specific binding and blocking experiments of 99Tc m-αCD8/Fab in vitro. SPECT/CT imaging was used to analyze the specific binding ability of the 99Tc m-αCD8/Fab probe in CT26 colon cancer mouse models (BALB/c). The near infrared fluorescence imaging and SPECT/CT imaging were performed to detect the intra-tumoral CD8 + T cell infiltration after anti-PD-1 therapy in tumor bearing mice, and the results were further verified by HE and immunofluorescence staining. CD8 + T cell depletion study was performed to determine the role of CD8 + T cells in the tumor responses to anti-PD-1 therapy. Two-way analysis of variance was used to compare the data difference. Results:The labeling yield of 99Tc m-αCD8/Fab was 90% with a high radiochemical purity (95%) and good stability in vitro (radiochemical purity still more than 80% after 720 min in PBS and FBS). 99Tc m-αCD8/Fab could specifically bind to mouse CD8 + T cells ((10.30±0.81) percent added radioactive dose (%AD)/10 6 cells), compared with the binding ability in human peripheral blood lymphocytes group and CD8 antibody blocking group ((1.78±0.61) and (1.59±0.25) %AD/10 6 cells; F=10.07, P<0.001). SPECT/CT imaging showed that 99Tc m-αCD8/Fab had markedly higher tumor uptake in the CT26 colon cancer mouse models. Near-infrared fluorescence imaging showed that the tumor uptake of 99Tc m-αCD8/Fab in the responsive group was significantly higher than in the nonresponsive group after anti-PD-1 treatment ((8.9±1.1)% vs (7.1±0.8)%; F=4.69, P=0.024), and SPECT/CT imaging found the similar result. HE and immunofluorescence staining of tumor and lymph nodes showed that the proportion of lymphocyte infiltration was higher in the responsive group. Furthermore, CD8 + T cell depletion significantly reversed the therapeutic effect of anti-PD-1 immunotherapy in tumor-bearing mice. Conclusions:In this study, 99Tc m-αCD8/Fab was successfully obtained. CD8-specific SPECT imaging could sensitively visualize the tumor-infiltrating CD8 + T cells, suggesting the potential application value to predict and evaluate the efficacy of immunotherapy in the clinical settings.

Establishment of a prediction model for colorectal cancer immune cell infiltration based on the cancer genome atlas (TCGA) database / 北京大学学报(医学版)

OBJECTIVE@#To study the correlation between immune cell infiltration in colorectal cancer tissue and clinical prognosis and to explore the levels of some immune cell genes for predicting the prognosis of patients with glioma colorectal cancer.@*METHODS@#In this study, we extracted colorectal cancer data from the cancer genome atlas (TCGA). Based on a deconvolution algorithm (called CIBERSORT) and clinically annotated expression profiles, the analysis assessed the infiltration patterns of 22 immune cells in colorectal cancer tissue to determine the association between each cell type and survival. Differences in five-year survival rate effectively illustrate the clinical prognostic value of each immune cell proportion in colorectal cancer, using a bar graph, correlation-based heatmap to represent the proportion of immune cells in each colorectal cancer sample.@*RESULTS@#A total of 473 colorectal cancer tissues and 41 normal control tissues were extracted from the TCGA database, and the comparative analysis showed that there were differences in the proportion of various TIICs in colorectal cancer tissues, which could characterize individual differences and have prognostic value. Among the cell subsets studied, the proportions of memory B cells, plasma cells, CD4+ T cells, natural killer (NK) cells, M0 macrophages, M2 macrophages, and activated mast cells were significantly different between normal and cancer tissues. Resting NK cells, CD8+ T cells, and plasma cells were associated with T phase, activated dendritic cells were associated with N phase, and eosinophils, M1 macrophages, and activated mast cells were associated with M phase. Survival analysis showed that activated dendritic cells were positively associated with five-year survival rate in colorectal cancer patients. Naive CD4+ T cells were inversely associated with five-year survival rate.@*CONCLUSION@#There are different degrees of immune cell infiltration in colorectal cancer tissues, and these differences may be important determinants of prognosis and treatment response. We conducted a new gene expression-based study of immune cell subtype levels and prognosis in colorectal cancer, which has potential clinical prognostic value in colorectal cancer patients.

Systematic evaluation of the relationship between tumor-infiltrating lymphocytes and prognosis of triple negative breast cancer / 国际外科学杂志

Objective:To investigate the prognostic value of tumor infiltrating lymphocytes (TILs) and their phenotypes, CD4 + TILs, CD8 + TILs and FOXP3 + TILs, in short-term prognosis (PCR) and long-term prognosis (DFS, OS) of triple negative breast cancer (TNBC), and to provide theoretical support for the immunotherapy of TNBC. Methods:A systematic surch of PubMed, Medline, Embase, Web of Science, CBM, CNKI was conducted to identified eligible articles. All prospective trials and observational controlled trials published before August 2020 were retrieved.The two authors independently evaluated the quality of the included literature, extracted the data, and conducted systematic evaluation and analysis using RevMan5.3 software.Results:A total of 48 literatures were included.Every 10% increase in TILs and TILs expression in the high-expression group predicted a higher PCR, OR were 1.81 (95% CI: 1.44-2.28) and 1.09 (95% CI: 1.02-1.15). For longer survival, the HR for DFS and OS were 0.99 (95% CI: 0.98-0.99) and 0.90 (95% CI: 0.86-0.94), 0.87 (95% CI: 0.79-0.96) and 0.88 (95% CI: 0.83-0.92). Phenotypic CD4 + TILs, CD8 + TILs, and FOXP3 + TILs did not show statistically significant differences in PCR among patients, but in long-term prognosis, higher infiltration predicted longer survival, with HR for DFS of 0.54 (95% CI: 0.36-0.80), 0.46(95% CI: 0.33-0.64) and 0.45(95% CI: 0.31-0.68). HR of OS were 0.49 (95% CI: 0.32-0.76), 0.59 (95% CI: 0.54-0.66) and 0.44 (95% CI: 0.27-0.71). Conclusion:The infiltration degree of TILs and the increase of TILs expression by 10% can be used as a prognostic index of TNBC.High levels of phenotypic infiltration of CD4 + TILs, CD8 + TILs, and FOXP3 + TILs predicted better long-term survival, but there was no statistically significant difference in short-term survival.

Expression and clinical value of glycogen synthase kinase-3β, Dickkopf-1 and β-catenin in proximal gastric cancer / 中国基层医药

Objective@#To study the expression and clinical value of glycogen synthase kinase-3β(GSK-3β), Dickkopf-1(DKK-1) and β-catenin in patients with proximal gastric cancer.@*Methods@#From April 2016 to April 2017, 47 patients with proximal gastric cancer in Chengfeng Hospital of Daqing Oilfield Group were enrolled in this study.Immunohistological tests were performed in all patients' lesions, adjacent tissues and healthy tissues.The effects of different factors on the expression of GSK-3β and DKK-1 were compared.@*Results@#The positive expression rate of GSK-3β in the normal tissues of patients with proximal gastric cancer was 70.21% (33/47), which was higher than those in the lesions and adjacent tissues[21.28% (10/47), 21.28%(10/47)](χ2=31.985, P<0.01). The positive expression rates of DKK-1 and β-catenin in the gastric cancer tissues were 38.30% (18/47), 82.98% (39/47), respectively, which were higher than those in the adjacent tissues and normal tissues[8.51% (4/47), 8.51% (4/47), and 6.38% (3/47), 2.13% (1/47)](χ2=20.517, 88.471, all P<0.01). The GSK-3β positive expression rate was higher in patients with tumor node metastasis (TNM) stage I-II, moderately well differentiated, infiltrated more than 50%, non-metastatic lymph nodes (P<0.05). The DKK-1 positive expression rate was higher in patients with TNM stage III-IV, moderately well differentiated and less than 50% infiltrated lymph nodes (P<0.05).@*Conclusion@#GSK-3β, DKK-1 and β-catenin are important indicators in the development and metastasis of gastric cancer.The above indicators have clinical value in the diagnosis and evaluation of curative effect.

Association between tumor-infiltrating lymphocytes and sentinel lymph node positivity in thin melanoma

Abstract: Background: Sentinel lymph node biopsy in thin invasive primary cutaneous melanoma (up to 1mm thick) is a controversial subject. The presence of tumor-infiltrating lymphocytes could be a factor to be considered in the decision to perform this procedure. Objective: To evaluate the association between the presence of tumor-infiltrating lymphocytes and lymph node metastases caused by thin primary cutaneous melanoma. Methods: Cross-sectional study with 137 records of thin invasive primary cutaneous melanoma submitted to sentinel lymph node biopsy from 2003 to 2015. The clinical variables considered were age, sex and topography of the lesion. The histopathological variables assessed were: tumor-infiltrating lymphocytes, melanoma subtype, Breslow thickness, Clark levels, number of mitoses per mm2, ulceration, regression and satellitosis. Univariate analyzes and logistic regression tests were performed as well the odds ratio and statistical relevance was considered when p <0.05. Results: Among the 137 cases of thin primary cutaneous melanoma submitted to sentinel lymph node biopsy, 10 (7.3%) had metastatic involvement. Ulceration on histopathology was positively associated with the presence of metastatic lymph node, with odds ratio =12.8 (2.77-59.4 95% CI, p=0.001). The presence of moderate/marked tumor-infiltrating lymphocytes was shown to be a protective factor for the presence of metastatic lymph node, with OR=0.20 (0.05-0.72 95% CI, p=0.014). The other variables - clinical and histopathological - were not associated with the outcome. Study limitations: The relatively small number of positive sentinel lymph node biopsy may explain such an expressive association of ulceration with metastatization. Conclusions: In patients with thin invasive primary cutaneous melanoma, few or absent tumor-infiltrating lymphocytes, as well as ulceration, represent independent risk factors for lymph node metastasis.

Clinical significance of tumor infiltrating lymphocytes in high-grade serous ovarian carcinoma / 中华病理学杂志

Objective@#To compare intratumoral tumor-infiltrating lymphocytes (TIL) in a cohort of high-grade serous ovarian carcinoma patients between those who treated with neoadjuvant chemotherapy (NACT) and primary debulking surgery (PDS), and to determine the clinical and prognostic significance of TIL in high-grade serous ovarian carcinoma.@*Methods@#Tissue microarrays and immunohistochemistry were used to assess the quantity of CD3+, CD4+ and CD8+ lymphocytes in tumor tissue from 138 high-grade serous ovarian carcinoma (PDS n=84, NACT n=54) which were collected from January 2013 to January 2016 at West China Second University Hospital, Sichuan University. TIL was analysed in two predefined groups of low and high TIL. The associations between clinical features and TIL were evaluated by χ2 test or Fisher′s exact test, and Kaplan-Meier survival analysis and Cox proportional hazards regression model were used for the association between the amounts of TIL and progression free survival.@*Results@#There was no difference in TIL/HPF (high-power field) counting in tumor tissue between PDS and NACT (P>0.05), and in the PDS cohort, CD3+, CD4+ and CD8+TIL were not associated with any clinical features like age, FIGO stage, tumor size and chemotherapy sensitivity, however, in the NACT cohort, CD8+TIL was strongly associated with chemotherapy sensitivity. The univariable analysis supported that high CD8+TIL in tumor tissue was associated with longer progression free survival both in the PDS and NACT cohort(P=0.030, P=0.032), but not CD4+TIL, in the NACT cohort, high CD3+TIL were also associated with longer progression free survival (P=0.019). Finally, in multivariate analysis, only the high CD8+TIL had prognostic significance (HR=0.369, 95%CI=0.176-0.772, P=0.008).@*Conclusion@#High CD8+lymphocytes density in tumor tissue was significantly associated with improved progression free survival in high-grade serous ovarian carcinoma. Besides, the CD8+lymphocytes density could be served as a potential marker of the chemotherapy sensitivity in patients who treated with neoadjuvant chemotherapy.

Association between p53 Expression and Amount of Tumor-Infiltrating Lymphocytes in Triple-Negative Breast Cancer

BACKGROUND: Most triple-negative breast cancers (TNBCs) have a high histologic grade, are associated with high endoplasmic stress, and possess a high frequency of TP53 mutations. TP53 missense mutations lead to the production of mutant p53 protein and usually show high levels of p53 protein expression. Tumor-infiltrating lymphocytes (TILs) accumulate as part of the anti-tumor immune response and have a strong prognostic and predictive significance in TNBC. We aimed to elucidate the association between p53 expression and the amount of TILs in TNBC. METHODS: In 678 TNBC patients, we evaluated TIL levels and expression of endoplasmic stress molecules. Immunohistochemical examination of p53 protein expression was categorized into three groups: no, low, and high expression. RESULTS: No, low, and high p53 expression was identified in 44.1% (n = 299), 20.1% (n = 136), and 35.8% (n = 243) of patients, respectively. Patients with high p53 expression showed high histologic grade (p < .001), high TIL levels (p = .009), and high expression of endoplasmic reticulum stress-associated molecules (p-eIF2a, p = .013; XBP1, p = .007), compared to patients with low p53 expression. There was no significant difference in disease-free (p = .406) or overall survival rates (p = .444) among the three p53 expression groups. CONCLUSIONS: High p53 expression is associated with increased expression of endoplasmic reticulum stress molecules and TIL influx.

Loss of Human Leukocyte Antigen Class I Expression Is Associated with Poor Prognosis in Patients with Advanced Breast Cancer

BACKGROUND: Human leukocyte antigen class I (HLA-I) molecules play important roles in regulating immune responses. Loss or reduction of HLA-I expression has been shown to be associated with prognosis in several cancers. Regulatory T-cells (Tregs) also play critical functions in immune response regulation. Evaluation of HLA-I expression status by the EMR8-5 antibody and its clinical impact in breast cancer have not been well studied, and its relationship with Tregs remains unclear. METHODS: We evaluated HLA-I expression and Treg infiltration by immunohistochemistry in 465 surgically resected breast cancer samples. We examined the correlation between HLA-I expression and Treg infiltration and clinicopathologic characteristics and survival analyses were performed. RESULTS: Total loss of HLA-I expression was found in 84 breast cancer samples (18.1%). Univariate survival analysis revealed that loss of HLA-I expression was significantly associated with worse disease-specific survival (DSS) (p = .029). HLA-I was not an independent prognostic factor in the entire patient group, but it was an adverse independent prognostic factor for DSS in patients with advanced disease (stage II–IV) (p = .031). Treg numbers were significantly higher in the intratumoral stroma of HLA-I–positive tumors than in HLA-I–negative tumors (median 6.3 cells/high power field vs 2.1 cells/high power field, p < .001). However, Tregs were not an independent prognostic factor in our cohort. CONCLUSIONS: Our findings suggest that the loss of HLA-I expression is associated with poor prognosis in breast cancer patients, highlighting the role of HLA-I alterations in immune evasion mechanisms of breast cancer. HLA-I could be a promising marker that enables the application of more effective and precise immunotherapies for patients with advanced breast cancer.

Expression of PD-1/PD-L1 in triple-negative breast carcinoma and its significance / 中华病理学杂志

Objective@#To investigate the correlation between the expression of PD-1, PD-L1 and clinicopathologic parameters in triple negative breast carcinoma (TNBC).@*Methods@#Samples from 151 patients with TNBC and 65 cases of other breast carcinomas (non-TNBC) were examined for PD-L1 and PD-1 expression by immunohistochemical staining on tissue microarray.@*Results@#The expression of PD-L1 and PD-1 in the tumor cells and interstitial lymphocytes in TNBC was significantly (P<0.05)higher than that in non-TNBC.In TNBC, the expression rates of PD-L1 in the cancer nests and stroma were 16.6%(25/151) and 25.2%(38/151)respectively.The former was positively correlated with tumor histological grade and lymph node metastasis (P<0.05), and the latter only with tumor histological grade (P<0.05). The expression rate of PD-1 was 27.2% (41/151), and correlated with PD-L1 expression in stromal tumor-infiltrating lymphocytes (P<0.05).@*Conclusion@#The expression of PD-L1 in TNBC is significantly correlated with high-grade histology, lymph node metastasis and PD-L1 expression in stromal tumor-infiltrating lymphocytes.

Correlation analysis of PD-L1 expression and prognosis in triple-negative breast cancers / 中华病理学杂志

Objective@#To investigate the relationship between PD-L1 expression and the clinicopathologic features and prognosis in triple-negative breast carcinomas (TNBC).@*Methods@#All 142 cases of TNBC were collected from the First Affiliated Hospital of Nanjing Medical University from February 2011 to December 2014, and the surgical excision or biopsy specimens from patients without chemotherapy and radiotherapy were included. Histopathologic analysis of stromal tumor infiltrating lymphocyte (sTIL) was performed on HE sections, and PD-L1 immunohistochemical staining was done with MaxVision.@*Results@#The PD-L1 expression rate was 34.5% (49/142) in tumor cells, and was 62.0% (88/142) in sTIL. The PD-L1 expression in tumor cells was positively correlated with tumor size (r=0.181, P=0.031), Ki-67 index (r=0.211, P=0.012), sTIL (r=0.380, P<0.01) and PD-L1 expression in sTIL (r=0.447, P<0.01). The PD-L1 expression in sTIL was positively correlated with tumor grade (r=0.215, P=0.01), Ki-67 index (r=0.253, P=0.002) and sTIL (r=0.370, P<0.01). The high stromal CD8+ /FOXP3+ ratio was significantly associated with improved overall survival (χ2=4.186, P=0.041). The high percentage of sTIL was significantly associated with improved overall survival (χ2=12.427, P<0.01) and progression-free survival (χ2=4.057, P=0.044).@*Conclusions@#In TNBC, PD-L1 expression is positively correlated with Ki-67 and sTIL; the stromal CD8+ /FOXP3+ ratio and sTIL are significantly associated with prognosis. The PD-L1 expression, stromal CD8+ /FOXP3+ ratio and sTIL are biologically important in TNBC, and all these correlative factors are important potential parameters in assessing immunotherapy for TNBC.

Identificação de subtipos moleculares baseada no perfil imunoistoquímico de carcinomas mamários triplo-negativos em mulheres com idade até 45 anos e sua distribuição nas diferentes regiões geográficas do Brasil / Identification of molecular subtypes based on immunohistochemical profile of triple-negative breast cancer (TNBCs) in women aged up to 45 years and its distribution in different geographical regions of Brazil

Carcinomas mamários triplo-negativos correspondem ao grupo heterogêneo de neoplasias mamárias caracterizadas pela ausência de expressão dos receptores de estrogênio e progesterona e sem amplificação ou superexpressão do HER2. São mais prevalentes em mulheres jovens e em afrodescendentes. Eles se associam frequentemente ao fenótipo basal-símile determinado geneticamente, entretanto, incluem também outros tipos moleculares intrínsecos. Metodologias de análise genética de novas gerações têm permitido sua estratificação em subgrupos distintos, o que justifica a heterogeneidade clínica deste grupo de neoplasias. A identificação desses subgrupos através de marcadores imunoistoquímicos de aplicação prática ainda é pouco explorada, embora seja uma ferramenta promissora na sua estratificação e determinação de alvos terapêuticos. OBJETIVOS: Nosso objetivo é explorar os perfis imunoistoquímicos dos carcinomas mamários triplo-negativos em mulheres com idade até 45 anos e investigar possíveis diferenças entre as cinco regiões geográficas brasileiras. MÉTODOS: Selecionamos 118 amostras de tumores de pacientes com idade até 45 anos, com carcinoma invasivo, blocos de parafina disponíveis e perfil imunoistoquímico triplo-negativo, procedentes das cinco regiões geográficas. Estes casos foram revisados quanto à determinação de tipo e grau histológico e as seguintes características anatomopatológicas: contorno do tumor, presença e fração do componente "in situ", embolização vascular peritumoral, tipo e grau da reação estromal, presença de necrose tumoral e formação de túbulos pela neoplasia. Foram selecionadas áreas representativas do tumor para construção de blocos de microarranjos de tecido para estudo imunoistoquímico. Foram pesquisados os seguintes marcadores: citoqueratinas basais 5/6 e 14, citoqueratinas luminais 8 e 18, receptor do fator de crescimento epidérmico (EGFR ou HER1), receptor de androgênio, e-caderina, catenina-beta,...

Triple-negative breast carcinomas (TNBC) correspond to a heterogeneous group of neoplasia characterized by the lack of expression of estrogen and progesterone receptors, and by the absence of amplification or overexpression of HER2. They are more prevalent among African descendants and younger women. They are often associated with the basal-like genetic phenotype; however, other intrinsic molecular types are included. Genomic analyses of next-generation methods have allowed stratification of TN breast carcinomas into distinct subgroups, explaining the clinical heterogeneity of this group of neoplasias. The identification of these subgroups by immunohistochemical markers is not well explored, although it represents a potential useful tool for the stratification and determination of therapeutic targets. OBJECTIVES: Our aim is to explore the TNBC immunohistochemical profile in patients of 45 year-old or younger, and to investigate possible differences among the five Brazilian geographic regions. METHODS: We've selected 118 samples of tumors from patients up to 45 years-old from five Brazilian geographic regions, with invasive carcinoma, available paraffin blocks, and triple-negative immunohistochemical profile. All the cases were reviewed as for determination of histologic type and grading, and the following pathological features: tumor contour, presence and percentage of in situ component, peritumoral vascular embolization, type and grade of stromal reaction, presence of tumoral necrosis, and tubule formation by neoplasia. Representative tumor areas were selected for tissue microarray construction for immunohistochemical study. The following markers were studied: Basal cytokeratins 5/6 and 14; luminal cytokeratins 8 and 18; Epidermal growth factor receptor (EGFR or HER1); androgen receptor; e-cadherin; catenin-beta; claudins (3,4 and 7); vimentin; smooth-muscle actin; p63, ALDH1, and Ki-67. According to the expression of the markers, the tumors...

Current Issues and Clinical Evidence in Tumor-Infiltrating Lymphocytes in Breast Cancer

With the advance in personalized therapeutic strategies in patients with breast cancer, there is an increasing need for biomarker-guided therapy. Although the immunogenicity of breast cancer has not been strongly considered in research or practice, tumor-infiltrating lymphocytes (TILs) are emerging as biomarkers mediating tumor response to treatments. Earlier studies have provided evidence that the level of TILs has prognostic value and the potential for predictive value, particularly in triple-negative and human epidermal growth factor receptor 2-positive breast cancer. Moreover, the level of TILs has been associated with treatment outcome in patients undergoing neoadjuvant chemotherapy. To date, no standardized methodology for measuring TILs has been established. In this article, we review current issues and clinical evidence for the use of TILs in breast cancer.

Association between Chemotherapy-Response Assays and Subsets of Tumor-Infiltrating Lymphocytes in Gastric Cancer: A Pilot Study

PURPOSE: The purpose of this pilot study was to evaluate the association between adenosine triphosphate-based chemotherapy response assays (ATP-CRAs) and subsets of tumor infiltrating lymphocytes (TILs) in gastric cancer. MATERIALS AND METHODS: In total, 15 gastric cancer tissue samples were obtained from gastrectomies performed between February 2007 and January 2011. Chemotherapy response assays were performed on tumor cells from these samples using 11 chemotherapeutic agents, including etoposide, doxorubicin, epirubicin, mitomycin, 5-fluorouracil (5-FU), oxaliplatin, irinotecan, docetaxel, paclitaxel, methotrexate, and cisplatin. TILs in the tissue samples were evaluated using antibodies specific for CD3, CD4, CD8, Foxp3, and Granzyme B. RESULTS: The highest cancer cell death rates were induced by etoposide (44.8%), 5-FU (43.1%), and mitomycin (39.9%). Samples from 10 patients who were treated with 5-FU were divided into 5-FU-sensitive and -insensitive groups according to median cell death rate. No difference was observed in survival between the two groups (P=0.216). Only two patients were treated with a chemotherapeutic agent determined by an ATP-CRA and there was no significant difference in overall survival compared with that of patients treated with their physician's choice of chemotherapeutic agent (P=0.105). However, a high number of CD3 TILs was a favorable prognostic factor (P=0.008). Pearson's correlation analyses showed no association between cancer cell death rates in response to chemotherapeutic agents and subsets of TILs. CONCLUSIONS: Cancer cell death rates in response to specific chemotherapeutic agents were not significantly associated with the distribution of TIL subsets.

The characteristics of lymph node metastasis and prognostic factors for carcinoma of papilla of Vater after pancreaticoduodenectomy / 中华肝胆外科杂志

Objective To analyze the characteristics ot lymph node metastasis and prognostic factors for carcinoma of papilla of Vater (CPV) after pancreaticoduodenectomy (PD).Methods From January 2005 to December 2010,94 patients with CPV underwent PD and dissection of regional lymph nodes at the Eastern Hepatobiliary Surgery Hospital.We carefully evaluated nodal involvement in the patients to determine the lymphatic spread of CPV and analyzed the clinicopathological variables in relation to prognosis.Results The overall rate of nodal involvement was 46.8％.Using the UICC staging (7th edition),lymphatic invasion in pT1,pT2,pT3 and pT4 were 15.4％ (2/13),62.7％ (32/51),80.0％ (8/10) and 100％ (2/2),respectively.The metastatic rates in the posterior pancreaticoduodenal lymph nodes,the mesopancreatic lymph nodes,the hepatoduodenal ligamental lymph nodes and the proper hepatic periarterial lymph nodes were 30.9％ (29/94),21.3％ (20/94),11.7％(11/94) and 6.4％ (6/94),respectively.Significant prognostic factors were tumor pT stage (P＜0.01),duodenal wall infiltration (P =0.001),liver metastasis (P =0.001),pancreatic paren chymal invasion (P=0.004),nodal involvement (P＜0.01) and different regional lymph nodes invasion (the posterior pancreaticoduodenal,P＜0.01; the mesopancreatic,P＜0.01; the hepatoduodenal ligamental,P＜0.01; the proper hepatic periarterial,P=0.010).Cox regression analysis for overall survival revealed that the posterior pancreaticoduodenal nodal involvement (P＜0.01),the mesopancreatic nodal involvement (P＜0.01) and duodenal wall infiltration (P=0.019) were significant independent prognostic risk factors.Conclusions The mesopancreatic lymph nodes and the posterior pancreaticoduodenal lymph nodes should equally be regarded as the first stop in lymphatic spread of CPV.Therefore,we should pay much attention to these regional lymph node dissections,especially to ensure complete resection of the uncinate process and the mesopancreas.

Significance of Foxp3 Positive Regulatory T Cell and Tumor Infiltrating T Lymphocyte in Triple Negative Breast Cancer

BACKGROUND: Triple negative breast cancer (TNBC) is defined as a lack of the expression of estrogen receptor, progesterone receptor and human epidermal growth factor receptor 2 in breast cancer. Many TNBCs show a profound infiltration of tumor infiltrating lymphocytes (TILs). It is still uncertain whether these TILs are protumoral or antitumoral. Regulatory T cells (Tregs) play a role in inducing immune tolerance to antigens, and they may be selectively recruited by cancer cells. This study was conducted to evaluate the significance of TILs with an emphasis on forkhead box p3 (Foxp3), which is a marker for CD25+CD4+ Treg in TNBC. METHODS: We investigated the Foxp3, CD8 and CD4 expressions in 100 cases of TNBC by immunohistochemistry and using a tissue microarray. The Foxp3 expression was divided as the high and low infiltration groups (cut-off value=20). RESULTS: The high infiltration group was correlated with higher histologic and nuclear grades. However, Foxp3+ Tregs were decreased in the T3 and T4 TNBCs as compared to that of the T1 and T2 TNBCs. No significant differences were found for the nodal status, lymphovascular invasion, stage, recurrence and overall survival. CONCLUSIONS: High Foxp3+ Treg infiltration in TNBC is correlated with the nuclear and histologic grades, but there was no relation to recurrence and overall survival.

Preoperative evaluation of brain invasion by diffusion tensor tractography in gliomas / 中国基层医药

Objective To evaluate the brain invasion in gliomas by diffusion tensor tractography(DTT).Methods Diffusion tensor imaging was preoperatively performed in 35 patients who histologically confirmed gliomas.13 of the 35 tumors were low-grade gliomas and 22 were high-grade gliomas. Then the spatial relationship between the lesions and white matter fiber tracts around tumor was analyzed. displacement, continuity and injured conditions of white matter fiber were observed. Results White matter fiber tract in all lesions could be observed clearly. Three patterns of white matter fibers involvement were identified:displaced,infiltrated and destructed. White matter fiber tracts around low-grade gliomas were primarily displaced ,but were mainly infiltrated and destructed around high-grade gliomas. Conclusion DTT was useful for showing white matter fiber tracts,observing the shape changes stereographically,and evaluating the relationship with gliomas in vivo.

Hepatocellular Carcinoma with Immature T-Cell (T-lymphoblastic) Proliferation

Indolent T-lymphoblastic proliferation has been rarely reported in the upper aerodigestive tract. The lymphoid cells associated with this condition have the morphological and phenotypical features of immature thymocytes. However, their pathogenesis and biology are unknown. We present an unusual type of tumor infiltrating lymphocytes in a case with hepatocellular carcinoma, presumed to be a T-lymphoblastic proliferation. A 58-yr-old female patient presented with indigestion and a palpable epigastric mass. The abdominal computed tomography revealed a mass in the S6 region of the liver. A hepatic segmentectomy was performed. Microscopic examination showed dense isolated nests of monomorphic lymphoid cells within the tumor. Immunohistochemically, the lymphoid cells were positive for CD3, terminal deoxymucleotide transferase (TdT) and CD1a. In addition, they showed dual expression of CD4 and CD8. The polymerase chain reaction used to examine the T-cell antigen receptor gamma gene rearrangement showed polyclonal T-cell proliferation. This is the second case of hepatocellular carcinoma combined with indolent T-lymphoblastic proliferation identified by an unusual tumor infiltrating lymphocytes.