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PURPOSE: This study sought to investigate associations among Yonsei criteria (tumor confined to the pancreas, intact fascia layer between the distal pancreas and the left adrenal gland and kidney, and tumor located more than 1–2 cm from the celiac axis) and tumor infiltrating lymphocytes in pancreatic cancer.MATERIALS AND METHODS: Patients who underwent curative distal pancreatectomy due to left-sided pancreatic cancer from January 2000 to December 2011 were enrolled. Follow-up was completed September 30, 2015.RESULTS: Fifty patients were enrolled. Having ≥ two metastatic lymph nodes (LNs, p=0.002), intraoperative transfusion (p=0.011), low levels of tumor infiltrating CD8⁺ T-cells (p=0.001), and a high Foxp3⁺/CD8⁺ ratio (p=0.009) were independent risk factors for disease-free survival. Not satisfying the Yonsei criteria (p=0.021), having ≥ two metastatic LNs (p=0.032), low levels of tumor infiltrating CD8⁺ T-cells (p=0.040) and a high Foxp3⁺/CD8⁺ ratio (p=0.032) were associated with unfavorable overall survival. High levels of CA19-9 and not satisfying the Yonsei criteria were significantly associated with a high Foxp3⁺/CD8⁺ ratio [Exp(β)=3.558; 95% confidence inverval: 1.000–12.658; p=0.050].CONCLUSION: Yonsei criteria may be clinically detectable biologic marker with which to predict immunologic status and survival in pancreatic cancer patients.
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Aim: Tumor-infiltrating lymphocytes (TILs) have a prognostic value in breast cancer (BC); however, because of the lack of standard evaluation methods, we aimed to assess the interobserver agreement of stromal TILs (sTILs) and intratumoral TILs (iTILs) as well as the effect of hot spot areas and molecular subtyping on the overall agreement. Methods: The study consisted of 121 haematoxylin and eosin (H and E)-stained slides of invasive BC samples obtained from the pathology archives. The TIL assessment was based on the International TIL Working Group recommendations for the percentage of sTILs and was conducted by four pathologists. The percentage of iTILs, the number of lymphocytes in hot spot areas (iTILs-HS), and the overall interobserver agreement for the molecular subtypes were evaluated. The interclass correlation coefficient (ICC) was used to assess interobserver agreement among the four pathologists. Results: The ICC score among the observers for the sTIL percentages was 0.74, and the individual ICC values for each molecular subtype were 0.55, 0.88, and 0.79 for luminal, HER2-positive, and triple-negative tumors, respectively. The compliance value for the iTILs was 0.29 (95% confidence interval (CI) = 0.06–0.48], whereas the compliance value for the iTILs-HS was 0.63 (95% CI = 0.49–0.71). The compliance values for the iTILs-HS subtypes were 0.72, 0.43, and 0.55 for luminal, HER2-positive, and triple-negative tumors, respectively. Conclusion: The IWTILG recommendations are reproducible and reliable. The interobserver agreement of the sTIL percentages was considerably higher for the triple-negative and HER2-positive cases than the luminal cases, whereas the interobserver agreement for the assessment of iTILs-HS in tumors was higher for the luminal subtype.
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Objective:To investigate the clinicopathological characteristics and immunohistochemical features of tumor-infiltrating lymphocyte(TIL) in ovarian serous adenocarcinoma,and their clinical value.Methods:The immunohistochemistry was used to observe 68 ovarian serous adenocarcinomacases'immunohistochemical features,and their correlations with clinicopathological characteristics were analyzed.Results:There were 43 cases with a large amount of TIL infiltrating in the 68 cases of serous adenocarcinoma,account for 63.24％.There were significant differences on the with or without a large amount of TIL infiltrating among the different tumor grade,different clinical stage,and different CA125 level (P ＜ 0.05).The numbers of CD3 +,CD4 + and CD8 + TIL in the cancer nest were significantly less than those in the stroma,with significantly statistical difference (P ＜ 0.05),Meanwhile the ratio of CD4 +/CD8 + in the cancer nest was significantly less than that in the stroma(P＜0.05).The ratio of CD4 +/CD8 + in the cancer nest and stroma at stage Ⅲ and with poor differentiation was significantly lower than in those at stage Ⅰ-Ⅱ and with good differentiation(P＜0.05).Meanwhile the ratio of CD8 +/FoxP3 + Treg in the cancer nest was less than that in the stroma(P＜0.05).The ratio of CD8 +/FoxP3 + Treg in the cancer nest at stage Ⅲ and with poor differentiation tumor was significantly lower than in those at stage Ⅰ-Ⅱ and with good differentiation(P＜0.05).The expression of GzmB in the cancer nest at stage Ⅲ and with poor differentiation tumor was significantly lower than that in the cancer nest at stage Ⅰ-Ⅱ and with good differentiation tumor(P＜0.05).Conclusions:With or without a large amount of TIL is related to tumor grade,clinical stage,and CA125 level inovarian serous adenocarcinoma.The numbers of TIL in ovarian serous adenocarcinoma increase considerably,but mainly in the stroma,and which may be related to several factors such as tumor cell differentiation and clinical stage.
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Objective The objectives of this study were to investigate the clinical significance of CD39,CD73,double positive subgroups for CD39 and CD73,and other lymphocytes with clinicopathological parameters in the microenvironment of colorectal cancer.Methods Tumor infiltrating lymphocyte(TIL)was collected from 24 patients with colorectal cancer after radical resection.The expression of CD39+,CD73+ or CD39+ with CD73+ in T cells were measured by flow cytometry.The association between these subgroups and clinicopathologic parameters was analyzed.Results The CD73+ and CD39+ with CD73+ subgroups were associated with lymph node metastasis and poor degree of differentiation,and this mechanism was closely related to tumor-associated inflammation.Conclusion CD39+ with CD73+ colorectal tumor infiltration Treg has a more unique biological activity than other Treg group.This study provides a new idea and theoretical basis for predicting the prognosis of colorectal cancer.
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Salmonella is a gram-negative bacterium that has an ability of tumor-targeting growth and proliferation. Attenuated Salmonella VNP20009 is a virulence genes-knockout bacterial strain based on Salmonella typhimurium, and it has an advantage of good therapeutic effect and low toxicity. One of the mechanisms of anti-tumor effect of VNP20009 is the induction of inflammatory reaction within tumor tissues. We used B16F10 melanoma model to investigate the mechanism of the anti-tumor effect of VNP20009. VNP20009 treatment effectively inhibited tumor growth and promoted the apoptosis and necrosis of tumor cells. VNP20009 increased the accumulation or infiltration of CD8+ T cells and CD11b+ monocytes within tumor tissue by raising the level of immune response and thus, induce the production of tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) to kill tumor cells by breaking the immuno-evasion barrier in the tumor microenvironment.
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Objective:To identify the signature of tumor-infiltrating lymphocyte (TIL) subtypes that may affect cytokine expres-sion between different outcomes of hepatocellular carcinoma (HCC) patients by analyzing the CD molecular expression profiles of non-cancerous hepatic tissues. Methods:Surface markers of TIL in noncancerous hepatic tissues from 146 HCC patients were determined by using immunohistochemical method and flow cytometry. Univariate and multivariate Cox proportional hazards models and Kaplan-Meier method were used to analyze the association of their expression levels with tumor recurrence and survival. Results:More than 86.4%of TILs in patients were quiescent, as measured via CD4+or Foxp3 expression. Meanwhile, more than 90%of CD3+T cells ex-pressed CD8+. The proportion of T cells was low compared with CD8+T cells. The proportion of CD19 and CD20 in distant nontumor tissues almost was zero. The proportion of T cell subgroups isolated from HCC circulating whole blood did not show a significant shift compared with the normal control, as follows:CD4+T/CD8+T=1.167 ± 1.04, CD8+T/CD3+T=0.288 ± 0.116, and CD4+T/CD3+T=0.429 ± 0.178. The proportion of CD8+T cells in noncancerous hepatic tissues was higher than that in blood (P<0.001).Conclusion:TILs in HCC noncancerous hepatic tissues are increased and contain a subpopulation of CD3+CD8+T cells. CD8+T cells in cancerous tissues, rather than noncancerous tissues, show significant differences between different prognostic groups.
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Objective To investigate the correlation of chemokine interleukin-8(IL-8) and tumor infiltrating lymphocyte.neutrophil in gastric cancer.Study the effect of chemokine IL-8 on local region immunocompetence and the mechanism of IL-8 in gastric cancer. Methods The distribution and quantitation of the subpopulations of tumor infiltrating lymphocyte including T_3、T_4、T_8 and neutrophil were studied with SABC immunhistochemistry and myeloperoxidase (MPO) kits respectively. Results Interleukin-8 and T_4/T_8 has a negative correlations (?=-0.52,P
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Pure choriocarcinoma is very rare in the testes, and host immune responses including tumor infiltrating lymphocytes are unusual in choriocarcinoma. This study reports a case of pure testicular choriocarcinoma with extensive lymphocytic infiltrate and granulomatous inflammation. Scrotal ultrasonography revealed a heterogeneous, hyperechoic intratesticular mass. -human chorionic gonadotropin levels were elevated in a radioimmunoassay. The hemorrhagic and necrotic solid mass was composed of two cell populations - mononuclear pleomorphic cells and intimately admixed multinucleated smudged cells. The tumor cells were positive for cytokeratin 7, epidermal growth factor receptors, human placental lactogen and p57. Many inflammatory cells were present within the tumor. The majority of infiltrating cells were CD8-positive cytotoxic cells, which also expressed granzyme-B and TIA-1. The tumor cells were positive for FasL, but negative for Fas. Therefore, this case seemed to escape the host defense response to the tumor due to the loss of Fas, although the cellular host immune response was still active.
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Male , Humans , Adult , Biomarkers, Tumor/analysis , Testicular Neoplasms/pathology , Lymphocytes, Tumor-Infiltrating/pathology , Granuloma/pathology , Chorionic Gonadotropin, beta Subunit, Human/metabolism , Choriocarcinoma/pathologyABSTRACT
Pure choriocarcinoma is very rare in the testes, and host immune responses including tumor infiltrating lymphocytes are unusual in choriocarcinoma. This study reports a case of pure testicular choriocarcinoma with extensive lymphocytic infiltrate and granulomatous inflammation. Scrotal ultrasonography revealed a heterogeneous, hyperechoic intratesticular mass. -human chorionic gonadotropin levels were elevated in a radioimmunoassay. The hemorrhagic and necrotic solid mass was composed of two cell populations - mononuclear pleomorphic cells and intimately admixed multinucleated smudged cells. The tumor cells were positive for cytokeratin 7, epidermal growth factor receptors, human placental lactogen and p57. Many inflammatory cells were present within the tumor. The majority of infiltrating cells were CD8-positive cytotoxic cells, which also expressed granzyme-B and TIA-1. The tumor cells were positive for FasL, but negative for Fas. Therefore, this case seemed to escape the host defense response to the tumor due to the loss of Fas, although the cellular host immune response was still active.
Subject(s)
Male , Humans , Adult , Biomarkers, Tumor/analysis , Testicular Neoplasms/pathology , Lymphocytes, Tumor-Infiltrating/pathology , Granuloma/pathology , Chorionic Gonadotropin, beta Subunit, Human/metabolism , Choriocarcinoma/pathologyABSTRACT
PURPOSE: Neoadjuvant chemotherapy for breast cancer creates new possibilities for the analysis of biological factors in the tumor and/or host, which may play a role in the response to treatment. In this study, whether changes in tumor infiltrating lymphocytes take place after neoadjuvant chemotherapy, and if these correlate with the response to treatment in breast cancer were analyzed. METHODS: Neoadjuvant chemotherapy (Adriamycin: 50 mg/m2 +docetaxel: 75 mg/m2, q3wks, 3 treatments) was followed by definitive surgical management. Histological sections from the pre-treatment core-needle biopsy and post-treatment surgical specimens of 17 patients were analyzed for the extent of lymphocytes infiltration. Infiltrated lymphocytes have been recognized as tumor-infiltrating lymphocytes (TILs; lymphocytes present within tumor cell nests). The modified Black's scoring system was used for grading the extent of lymphocytes infiltration, with immunohistochemical (IHC) staining used to characterize the TILs. RESULTS: Pre-treatment lymphocytic infiltrates within the tumor were minimal in the majority of patients, and showed no relationship with the response. A marked increase of TILs after chemotherapeutic treatment was noted in patients according to the following response; complete response: 4/4 (100%), partial response: 2/6 (33.3%), stable disease: 0/7 (0%) (P=0.004). Histological sections stained with IHC staining revealed the increased TILs were CD8 positive cytotoxic T-lymphocytes. CONCLUSION: These results suggest that the development of TILs after treatment correlate with the response to neoadjuvant chemotherapy. Despite the limitation of this preliminary study, the extent of TILs change might be used as a predictor for the therapeutic efficacy of neoadjuvant chemotherapy in breast cancer.
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Humans , Biological Factors , Biopsy , Breast Neoplasms , Breast , Drug Therapy , Lymphocytes , Lymphocytes, Tumor-Infiltrating , T-Lymphocytes, CytotoxicABSTRACT
Objective:To select an appropriate dosage of IL-2 for tumor infiltrating lymphocyte(TIL).Methods:Isolated by the attachment method we established,TIL was cultivated in self-supernatant of malignant pleura,with three different concentrations of IL-2,such as 6000u per ml,6000u per ml for the first administration followed by 1000u per ml,or 1000u per ml.The expansion,killing activity and phenotype changes of TIL cultured in different cultures were assessed.Results:As cultured in self-supernatant of malignant pleura fluid,the concentrations of IL-2,such as 6000u per ml or 6000u per ml for the first administration followed by 1000u per ml seemed benefit for TIL proliferation.Conclusion:6000u per ml of IL-2 for the first administration was very important.It could help TIL to activate and proliferate early.The study described here offers the possibility for TIL cultured in vitro self-supernatant of malignant pleura fluid in vitro for a short time,and then reinfuse to thorax for further expanding and controlling the malignant pleura effusion in the presence of IL-2,and thereby minimizing the risks of contamination.
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Objective:To study the effect of autologous supernatant of malignant pleura effusion and RPMI1640 with 10% AB+ serum on the culture of tumor infiltrating lymphocytes (TIL) in vitro. Methods:Isolated by the attachment method we established, TIL was cultured in either autologous supernatant of malignant pleura fluid or RPMI1640 with 10% AB+ serum, and various types of cytokines such as (IL-2,) PHA and antibody against CD3 (OKT3) were added into both cultures. The proliferation as well as the killing activity and the phenotype changes of TIL cultured in the two kinds of cultures were compared. Results:There was no difference in the proliferation or the killing activity in vitro as well as the phenotype changes of TIL between the two kinds of cultures. Conclusion: Autologous supernatant of malignant pleura fluid could be used as TIL culture medium. The method described here made it possible for TIL to be cultured in vitro for a short time, and then reinfused back to thorax for further expanding and controlling the malignant pleura effusion in the presence of IL-2, and it alsominimized the risks of contamination.
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0.05) except that IFN-? secretion of L-SEA group was lower than that of SEA group at 4th day (P
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Objective To investigate the killing activity of tumor infiltrating lymphocytes (TIL) stimulated by dendritic cells (DC) on gastric carcinoma cells in vitro. Since DC is the strongest antigen presenting cell (APC) which has been known, it can present antigens to T lymphocytes, including TIL, and can induce cytotoxic T lymphocyte (CTL) responded in vivo and vitro. Methods DC were isolated from the peripheral blood of patients with gastric carcinoma and stimulated by granulocyte/macrophage colony stimulating factor (GM CSF), interleukin 4 (IL 4) and tumor antigen. Then TIL were stimulated by DC and their killing activity on autogenous gastric carcinoma cells and SGC 7901 cells in vitro were observed. Results TIL stimulated by DC had very high killing activity on autogenous gastric carcinoma cells and the killing rate was (89.39?3.05)%, which was much higher than those of TIL not stimulated by DC and T lymphocytes stimulated by DC as well as T lymphocytes not stimulated by DC on autogenous gastric carcinoma cells. The killing rates of the last three ones were (54.37?1.50)%, (53.92?1.46)% and (3.55? 0.25) % respectively. However, their killing activity on SGC 7901 cells was lower. Conclusions The results indicate that DC from patients with gastric carcinoma can induce efficient and specific anti gastric carcinoma immune responses.
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Objective To study the effects of staphylococcal enterotoxin B(SEB) on the proliferation of tumor infiltrating lymphocytes(TILs) from rectum adenocacinoma.Methods The TILs from patients with rectum adenocacinoma were stimulated with SEB and interleukin 2(IL-2) respectively,and then the proliferation of TILs,the secretion of IL-2 and tumor necrosis factor-?(TNF-?) were determined.Results SEB presented profound stimulating effect on the TILs from rectum adenocarcinoma both the proliferation of TILs and the secretion of cytokines.Compared with the IL-2,SEB stimulated TILs more quickly,and SEB acted more effectively in the early stage but weakly in the late stage.Conclution SEB was an effective TIL stimulator.
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PURPOSE: This study was to observe whether the apoptotic function of tumor-infiltrating lymphocytes (TIL) is induced in human gastric epithelial dysplasia and gastric adenocarcinoma according to the role of FasL expression. MATENRIALS AND METHODS: A total of 56 gastric epithelial dysplasia and gastric adenocarcinoma patients were enrolled in this study: 9 cases of gastric epithelial dysplasia, 18 cases of early gastric carcinomas (EGC) and 29 cases of advanced gastric carcinomas (AGC). Immunohistochemical staining was performed for FasL and CD45, and the terminal deoxynucleotidyl transferase mediated dUTP nick end labelling (TUNEL) method was used to detect cell death in tumor-infiltrating lymphocytes. RESULTS: 1) Positive reactions of FasL to neoplastic cells were 88.9% (8/9) in gastric epithelial dysplasia, 83.3% (15/18) in EGC, and 75.9% (22/29) in AGC. 2) Expression of TIL was decreased in the FasL positive region and was increased in the FasL negative region, and significant expression of TIL was observed in the AGC group (P=0.001). 3) Expression of apoptotic TIL was very similar to the FasL expression, and 100% expression was observed in gastric epithelial dysplasia group. 4) Expression of apoptotic TIL was increased in the FasL positive region and decreased in the FasL negative region, and significant apoptotic expression was observed in the gastric epithelial dysplasia and EGC groups (P=0.0420, P=0.0263, respectively). CONCLUSION: These results suggest that FasL is a prevalent mediator of immune privilege in epithelial dysplasia and cancer of the stomach.
Subject(s)
Humans , Adenocarcinoma , Apoptosis , Cell Death , DNA Nucleotidylexotransferase , Lymphocytes, Tumor-Infiltrating , Stomach NeoplasmsABSTRACT
PURPOSE: The purpose of this study was to determine whether Fas-L expression is associated with increased apoptotic induction of tumor-infiltrating lymphocytes (TIL) in human gastric carcinomas. MATERIALS AND METHODS: The author analysed 38 cases of early gastric carcinoma (EGC) and 61 cases of advanced gastric carcinoma (AGC) who received gastric resection, in whom the number of diffuse type was 38 cases and the number of intestinal type was 61 cases. The author used immunohistochemical staining for Fas, Fas-L and CD45, and TUNEL in situ apoptosis detection kit. TIL were detected by CD45 and apoptosis of TIL were detected by CD45 expression and TUNEL positivity on serial histologic sections. RESULTS: Fas-L was localized to neoplastic cells in 61% (23/38) of EGC group and 66% (40/61) of AGC group. The extent of Fas-L expression was variable, with both Fas-L positive and negative neoplastic region occuring within tumors. TIL adjacent to Fas-L expressing tumor region were decreased in number and TIL adjacent to FasL-negative tumor region were increased in number; apoptotic induction of TIL showed just the opposite pattern (p<0.05). Fas expression was found essentially homogeneously throughout the tumor mass independent of tumor stage. Fas expression showed 64% (39/61) of intestinal type and 68% (26/38) of diffuse type. Labeling indices for tumoral apoptosis in EGC and AGC were 6.72% and 7.13%, respectively and this difference was statistically insignificant. Co-expression of Fas-L and Fas, which occurred over large areas of the tumors, did not result in an enhanced rate of tumor cell apoptosis. In addition, factors such as tumor stage and other prognostic factors were not concerned in Fas and Fas-L expression, number of TIL and apoptotic induction. CONCLUSION: These findings suggest Fas-mediated apoptotic depletion of TIL in response to Fas-L expression by stomach cancers, and provide the evidence to support the Fas counterattack as a mechanism of immune escape in gastric cancer. In addition, gastric carcinoma cells of the intestinal and diffuse type did not differ in their expression of the apoptotic receptor Fas.
Subject(s)
Humans , Apoptosis , In Situ Nick-End Labeling , Lymphocytes, Tumor-Infiltrating , Stomach Neoplasms , United NationsABSTRACT
PURPOSE: To increase the potency of TIL(tumor infiltrating lymphocytes) in immunotheraphy, the effect of TNF(tumor necrosis factor) and IL-2(interleukin-2) were investigated in vitro on proliferation and cytotoxic effect of TIL to the various kinds of tumor lines and fresh frozen tumor cells from renal cell carcinoma patient. MATERIALS AND METHOD: For the target cell of the TIL cytotoxicity was used M-l 4(Melanoma tumor line) for investigation of LAK activity and erythroleukemic cell line(K562) for investigation of natural killer(NK) cell activity. To investigate the nonspecific cytotoxicity on renal cell carcinoma, autologous fresh frozen renal carcinoma cells were used as the autologous target, and allogeneic fresh frozen renal carcinoma cells and renal cell carcinoma line(444) as the allogeneic target cells. The dosage of TNF was 500univm1 and IL-2 was 1000Cetus unit/ml. The proliferation effect of TIL was checked by {H}Thymidine uptake assay and its cytotoxicity was checked with 4 hour Chromium release cytotoxicity assay. RESULTS: The proliferation erect of TIL, treated with IL-2 at 20ays of culture was 11486+/-591cpm and 23817+/-1069cpm with combination of IL-2 and TNF. The nonspecific cytotoxicity of TIL for the 444 at 20th day of culture was 8.2Lytic unit(LU) in IL-2 only and 18,5LU with combination of 1L-2 and TNF. The cytotoxicity of IL for M-14 at 20th day of culture was 8.2LU in IL-2 only and 19.0LU with combination of IL-2 and TNF. The cytotoxicity of TIL for the K562 at 20th day of culture was 37.3LU in IL-2 only and 60.4LU with combination of IL-2 and TNF. CONCLUSIONS: These results suggest that the combination of TNF and IL-2 has a synergistic effect on proliferation and nonspecific cytotoxicity of TIL at 20 6ays of culture in vitro. These results bear important practical implication for clinical trials of TIL in adoptive immunotherapy.
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Humans , Carcinoma, Renal Cell , Chromium , Immunotherapy, Adoptive , Interleukin-2 , Lymphocytes, Tumor-Infiltrating , Necrosis , Tumor Necrosis Factor-alphaABSTRACT
Objective: To investigate the effects of dendritic cells(DCs) loaded with tumor antigens on the proliferation and antitumor activity of tumor-infiltrating lymphocytes(TILs) from hepatocellular carcinoma (HCC) in vitro . Methods:DCs derived from the peripheral blood of patients with HCC were loaded with autologous tumor lysate(Tuly) to generate DC-Tuly and then the DC-Tuly was co-cultured with autologous TILs in low concentration of IL-2. TILs were counted, and the phenotypes of DCs and TILs were assayed by FCM and the cytotoxicity of TILs was determined with LDH method. The concentration of cytokines in TILs culture supernatant such as IFN-? and TNF-? was detected by ELISA . Results: The expression levels of CD1a, CD80, CD83,CD86, CD40, HLA-DR molecules on the DC-Tuly are notably increased( P
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We selected defective retrovirus LXSN approved by the recombinant DNA Advisory Comittee, NIH, USA as a vector to study the transduction of human TIL with TNF gene, on the basis of which, we engaged in the clinical application. The results showed that the logarithmic growth phase was the optimal transducing time when the TIL were isolated and cultured from 9 to 25 days. It also showed that the transduction efficiency was correlated to the MOI value, the more MOI value, the more TIL being transduced. Successful gene inserted and expressing were confirmed by the polymerase chain reaction and the L929 cell test. The clinical trial of treatment for metastastic hepatic cancer suggested that the better therapeutic effectiveness and less side effect depend on the specificity of the TIL, the proper infusion pathway and the dose of the transduced TIL.