MHC-dependent cytolysis of autologous tumor cells by lymphocytes infiltrating urothelial carcinomas

Tumor-infiltrating lymphocytes (TIL) were grown from 23 urothelial carcinomas. Phenotyping analysis showed that the TIL cultures were mainly CD3⁺. Although CD4⁺ and CD8⁺ T-cell sub-sets were grown in culture, CD4⁺ T-cell sub-sets predominated over CD8⁺ T cells. Immunohistochemical studies performed on 5 tumor specimens confirmed this observation, and indicated that CD4⁺ T cells surrounded the tumor islets, whereas CD8⁺ T lymphocytes were localized among the tumor cells. Five short-term carcinoma cell lines established from these urothelial tumors were used as target cells in cytolysis assays in order to investigate the functional anti-tumor activity of autologous TIL. TIL from 4/5 tumors were lytic and 3 TIL lines displayed MHC-class-I-dependent cytotoxicity directed against autologous tumor cells. CD4⁺ T-cell-depletion experiments performed on TIL line 07 confirmed that CD8⁺ MHC-class-I-dependent CTL were the predominant effectors. Finally, experiments performed on 6 allogeneic urothelial-cancer cell lines matched for HLA-class-I molecules showed that TIL07 exhibited selective lytic activity toward tumor 07. These data indicate that CD8⁺ MHC-class-I-dependent CTL present in urothelial carcinomas are functional and may participate in the anti-tumor immune response. Int. J. Cancer 71:585-594, 1997. © 1997 Wiley-Liss, Inc.     

Study of tumor infiltrating lymphocytes and transforming growth factor-β as prognostic factors in breast carcinoma

Cytokines and growth factors are powerful modulators of the immune response. Their aberrant expression either by the tumor cells or by the tumor infiltrating lymphocytes confers a selective advantage to the tumor to grow and suppress the cytotoxic activity of the infiltrating lymphocytes. Therefore, analysis of these soluble factors in the tumor microenvironment can provide an insight into the understanding of the tumor behavior and may be used as a prognostic factor. In the present study the nature of the tumor infiltrating lymphocytes (TILs) and cytokine profile was examined in 36 and 19 mammary carcinoma tissues, respectively, by immunohistochemistry and PCR. Phenotypic differences in the number of cytotoxic T lymphocytes (CD8⁺) and lymphokine activated killer cells (CD16) was observed among TILs when patients with either early disease stage (39% and 46.6%, respectively) or those alive with no residual disease (31% and 52%, respectively) were compared with late stage (9.7% and 22.8%, respectively) or those dead of disease (14.6% and 15.6%, respectively). Furthermore, analysis of the 19 tumor samples for cytokine mRNA expression by RT-PCR revealed the presence of TNF-α, IL-10, TGF-β1, and IL-2. However, semi-quantitative PCR analysis demonstrated TGF- β1 expression to be significantly higher in patients with a favorable outcome (1.0246 attomoles/μmoles) as compared to patients with a poor prognosis (0.1157 attomoles/μmoles). Our results demonstrate the potential biological significance of certain host factors, particularly TILs and TGF β1 expression, on the outcome of breast cancer. Int. J. Cancer 74:492–501, 1997. © 1997 Wiley-Liss, Inc.     

Programmed death ligand‐1 is associated with tumor infiltrating lymphocytes and poorer survival in urothelial cell carcinoma of the bladder

Drugs blocking programmed death ligand‐1 (PD‐L1) have shown unprecedented activity in metastatic and unresectable bladder cancer. The purpose of the present study was to investigate the expression, clinical significance and association of PD‐L1 with tumor‐infiltrating lymphocytes (TIL) in resectable urothelial cell carcinoma of the bladder (UCB). In this retrospective study, 248 UCB patients who received radical cystectomy or transurethral resection were examined. Immunohistochemistry was used to evaluate PD‐L1 expression and stromal CD8⁺ TIL, Th1 orientation T cell (T‐bet⁺) and PD‐1⁺ TIL densities within the intratumoral regions and associated stromal regions. Of the 248 specimens, 23% showed PD‐L1 expression in tumor cells and 55% in tumor‐infiltrating immune cells. CD8⁺ TIL, T‐bet⁺ TIL and PD‐1⁺ TIL were distributed throughout the tumor tissues and were more frequently distributed in stromal regions than in intratumoral regions. PD‐L1⁺ tumor cells and PD‐L1⁺ immune cells were positively associated with aggressive clinical features (all P < .05). Both PD‐L1⁺ tumor cells and PD‐L1⁺ immune cells were associated with poorer recurrence‐free and overall survival (all P < .05). Multivariate analysis showed that PD‐L1⁺ immune cells were an independent prognostic factor for overall (P = .001) and recurrence‐free survival (P = .024). Notably, high stromal CD8⁺ TIL and PD‐1⁺ TIL density were associated with poorer overall survival (P = .031 and P = .001, respectively). In the stroma, CD8⁺ TIL density has strong positive association with PD‐L1⁺ immune cells and PD‐1⁺ TIL density (all P < .0001). These results suggested that an exhausted immune state occurred in the tumor stroma in UCB. Further clinical development of immune‐checkpoint inhibitors may be effective for resectable patients with UCB.     

肝癌肿瘤浸润淋巴细胞抗瘤活性及表型特征

目的 探讨肝癌肿瘤浸润淋巴细胞的抗瘤活性及表型特征。方法 13例肝癌患者(7例接受术前TACE，6例未接受)，MTT法检测TIL细胞抗肿瘤活性，APAAP法检测TIL细胞表型。结果 TACE组TIL细胞在第2周增殖达到高峰，至第3周平均扩增52倍，而未接受术前化疗组在第3周扩增达到高峰，平均扩增了12倍。术前介入化疗患者TIL细胞肿瘤杀伤活性增强，CD3^ 、CD4^ 、CD8^ 不同程度的增加。结论 术前介入化疗可缩短肝癌患者TIL细胞增殖时间，提高肿瘤杀伤活性。     

肿瘤浸润淋巴细胞的体外培养及临床应用研究

目的:研究肺癌患者胸腔积液中肿瘤浸润淋巴细胞(tumorinfiltratinglymphocytes,TIL)对瘤细胞的杀伤活性及其表型变化,并对其回输后毒副反应及患者免疫功能进行观察。方法:应用贴壁法分离恶性胸腔积液中TIL,并经rIL2诱导培养;用间接免疫荧光法测定CD3+、CD4+和CD8+比例;应用3HTdR释放法测定TIL对自体瘤细胞和801D细胞的杀伤作用;用活化的自体TIL注入患者胸腔内,观察胸腔积液变化。结果:第21天时CD3+和CD8+比例明显提高,而CD4+比例和CD4+/CD8+比值变化不大;对自体瘤细胞和801D细胞的杀伤力明显提高;40例自体回输治疗胸腔积液的患者,有效率为72.5%,且一般状况好,毒副反应小,辅助检查无异常改变,免疫功能增强。结论:肺癌TIL过继免疫治疗对恶性胸腔积液患者是一种安全、有效的免疫治疗方法。     

COX-2 expression in canine and feline invasive mammary carcinomas: correlation with clinicopathological features and prognostic fmolecular markers

Summary Cyclooxygenase (COX)-2 is an inducible enzyme linked to tumor growth and angiogenesis. Its expression occurs in a wide range of preneoplastic and neoplastic conditions in humans, including colon and breast carcinomas. We evaluated the role of COX-2 as a mediator of angiogenesis in feline and canine invasive carcinomas (IMCs) and its role as a prognostic indicator. COX-2 expression was assessed in neoplastic samples and healthy mammary glands by immunohistochemistry, and related to the following clinicopathological parameters: age, tumor size, histologic type, tumor grading, vessel invasion, estrogen (ER) and progesterone receptor (PR) status, Ki-67, HER-2 overexpression, microvessel density (MVD), VEGF expression and overall survival (OS). In both species, COX-2 immunoreactivity was not observed in healthy tissues, whereas 96% of feline and 100% of canine invasive carcinomas scored positive. In queens, COX-2 overexpression was significantly correlated to ER-negative status (p=0.04) and to increased PR (p=0.038) expression, and angiogenesis assessed by VEGF expression (p=0.002). In bitches an increased COX-2 expression was significantly correlated to HER-2 overexpression (p=0.013) and to tumor dedifferentiation (p=0.03). In both species increased levels of COX-2 were correlated to poorer prognosis (p=0.03 in dogs and p=0.002 in cats). COX-2 is expressed in mammary tissues during tumorigenesis and its expression is associated with a poorer prognosis in bitches and queens. The correlation of COX-2 expression and angiogenesis provides support for a potential role of COX-2 inhibitors for the prevention and the treatment of feline IMCs via their anti-angiogenic properties. In the canine species, moreover, COX-2 may be important for mediating HER-2 induced mammary tumors.     

New prognostic factors associated with long-term survival in node-negative breast cancer patients

Background  This study was undertaken to determine the absolute and relative value of angiogenesis, proliferating cell nuclear antigen (PCNA) and conventional prognostic factors in predicting relapse-free survival (RFS) and overall survival (OS) rates associated with long-term survival in Japanese patients with node-negative breast cancer. Patients and Methods  Two hundred patients with histological node-negative breast cancer were studied. We investigated nine clinicopathological factors, including angiogenesis, PCNA using permanent-section immunohistochemistry, clinical tumor size, histological grade (HG), tumor necrosis, lymphatic vessel invasion (LVI), histological extension, histological classification, and infiltrating growth (INF), followed for a median of 10 years (range, 1 to 20). Results  Twenty-one patients (10.5%) had recurrence and 15 patients (7.5%) died of breast cancer. Univariate analysis showed that PCNA, clinical tumor size, HG, angiogenesis, and LVI were significantly predictive of 20-year RFS or OS. Tumor necrosis was significantly predictive of OS, not of RFS. Multivariate analysis showed that clinical tumor size (P=0.0003), angiogenesis (P=0.0003), PCNA (P= 0.0064), and HG (P=0.0401) were significant independent prognostic factors for RFS. PCNA (P< 0.0001) and clinical tumor size (P=0.0112) were significant independent prognostic factors for OS, while angiogenesis was a borderline significant factor. Conclusion  PCNA and angiogenesis were important new prognostic factors in node-negative breast cancer patients.     

Anaplastic oligodendrogliomas: prognostic factors for tumor recurrence and survival

OBJECTIVES: Anaplastic oligodendrogliomas (AO) are uncommon primary brain tumors whose natural history, prognosis, and optimal management are not yet fully understood. However, they are associated with a better prognosis and response to multimodality therapy based on specific molecular changes. In this multicenter retrospective study, we analyzed the clinical characteristics of patients with AO to identify prognostic factors that influence time to progression (TTP) and survival. METHODS: A retrospective search of the brain tumor databases of three institutions (the University of Texas M. D. Anderson Cancer Center, the University of California at San Francisco, and the University of Utah) for patients between 1977 and 1995 with histologically confirmed AO identified a cohort of 106 patients that was further analyzed in this study. Initial treatment included surgery alone (n = 12) or surgery followed by one of the following: radiotherapy (RT) alone (n = 49), chemotherapy alone (n = 4), chemotherapy followed by RT (n = 10), RT followed by chemotherapy (n = 20), and others (n = 11). RESULTS: The median age at diagnosis was 43 years, and the median Karnofsky performance score (KPS) was 90. The overall median survival was 7.3 years, and the 5-year survival rate was 62%. Univariate analysis of several clinical variables showed that only age (p < 0.0001) and KPS (p = 0.04) correlated significantly with survival. Fifty patients had disease progression after initial therapy. The median TTP was 48 months. Age at diagnosis was the only variable that correlated significantly with TTP. CONCLUSIONS: A trend towards longer survival with a greater extent of resection was evident. The relative efficacy of various treatment modalities could not be definitively determined because of the heterogeneity of the therapies used. Overall, patients with AO have a better prognosis after therapy compared with those who have other malignant gliomas.     

肿瘤浸润淋巴细胞对不同分子亚型乳腺癌预后效应的研究进展

肿瘤浸润淋巴细胞（tumor infiltrating lymphocytes,TILs）是肿瘤微环境的重要组分,对结肠癌、肺癌等多种肿瘤预后有明确的指示效应。在乳腺癌中,相关研究的结论并不一致。考虑到乳腺癌是一组由多种不同分子亚型组成的异质性疾病,TILs对不同分子亚型乳腺癌的预后指示作用也不同。本综述阐述常用TILs指标对不同分子亚型乳腺癌预后指示效应的相关研究进展。     

胃肠道间质瘤的临床病理特征和预后相关因素分析

目的探讨胃肠道间质瘤（gastrointestinal stromal tumor，GIST）的临床病理特征和预后相关因素，评价Fletcher等推荐的GIST生物学行为分级法的应用价值。方法复阅切片，以免疫组化方法检测CD117、CD34和S—100蛋白的表达。结合患者临床病理特征和GIST生物学行为分级分析影响GIST患者预后的相关因素。结果确诊44例GIST患者。患者中位生存时间41．7个月，1、3和5年总体生存率分别为80．2％、57．2％和39．8％。单因素分析显示，肿瘤最大径〉5cm、肿瘤性坏死、组织学类型混合型、肿瘤细胞高密集度、核分裂象计数≥5／50HPF、手术未完整切除、复发或转移瘤以及浸润性生长显著降低生存（P〈0．05）。44例GIST生物学行为分级：极低度侵袭危险性7例，低度侵袭危险性8例，中度侵袭危险性10例，高度侵袭危险性19例。单因素分析显示，不同侵袭危险性组患者生存率差异有显著性（X^2=28．376，P=0．000）。结论准确判断GIST患者预后有赖于综合分析患者临床病理特征。Fletcher等推荐的GIST生物学行为分级法能够准确评价GIST患者的预后，具有较好的临床应用价值。     

Immunologic character of tumor infiltrating lymphocytes in ovarian carcinoma

Tumor-infiltratinglymphocytes(TIL)wasdirectlyisolatedfrompatient'stumortissues.ByrIL2activationandexpansioninvitro,TILwasagainimportedintothesamepatient'sbodytotreattumor,withapparenteffectsofanti-tumorandcomparativelessside-effect,withoutkillingothertumorcellsandnormalcells,allofwhichhavemadeitaeffectivewayoftreatingtumoratanadvancedstage.Inrecentyears,someresearchersfoundthattheeffectsofTIL'treatmentwerenotsofarapparent,withcomparativelyapparentdifference,whichmightbecloselyrelatedtothef…     

肿瘤浸润淋巴细胞的抗肿瘤免疫困境

体外肿瘤浸润淋巴细胞(TIL)具有一定的抗瘤活性,过继免疫治疗却未表现出理想效果,研究显示恶性肿瘤患者免疫系统处于系统性缺陷或耐受状态.在诱发阶段树突状细胞(DC)诱导TIL发生免疫偏倚,有的则因DC严重不足或状态不佳使TIL几无功能.肿瘤细胞的生理变化及肿瘤局部微环境也使TIL的杀伤活性受到严重抑制,甚至被反杀伤.体内TIL自身变化同样造成抗瘤免疫力低下.因此TIL的抗瘤免疫受到多方面困扰.     

General features of gastric carcinomas and comparison of HSP70 and NK cell immunoreactivity with prognostic factors

During the period of 1996-1998 ninety-four gastrectomy specimens with gastric carcinoma referred to Erciyes University, Medical Faculty, Department of Pathology, were examined histopathologically, histochemically and immunohistochemically. General characteristics of gastric carcinomas and prognostic factors were studied. According the Lauren classification, of the 94 cases of gastric carcinomas, 56 were intestinal type, 21 were diffuse type and 17 were mixed type carcinoma. The association rates of Helicobacter pylori, chronic atrophic gastritis and intestinal metaplasia with gastric carcinomas were high. There was strong immunorectivity with HSP70 in 62,5% of the intestinal type carcinomas. This ratios were lower in diffuse and mixed type carcinomas (p<0.05). The more tumor size and invasion depth increased, the more HSP70 immunoreactivity was obtained (p<0.05). HSP70 immunorectivity was considerably higher in the patients having lymph node metastasis and vascular invasion (p<0.05). It was found that the NK cell number was low in the tumor but higher around the tumor in early gastric carcinomas, compared with advanced carcinomas (p>0.05). In the tumors larger than 10 cm with vascular invasion, NK cell number was lower around the tumor (p>0.05). Defining prognostic factors of gastric carcinomas is of importance to clinicians. It is thought that HSP70 immunoreactivity, besides invasion depth, lymph node metastasis, vascular invasion, tumor size and inflammatory reaction against the tumor, is important in prognosis and associated with advanced stage.     

肿瘤浸润淋巴细胞与lncRNA在肿瘤发展中作用的研究进展

肿瘤浸润淋巴细胞在肿瘤微环境中扮演着重要的角色,不同的细胞亚型在肿瘤的进展中发挥抗肿瘤或促肿瘤作用.肿瘤浸润淋巴细胞主要包括T淋巴细胞、B淋巴细胞和自然杀伤细胞,这些细胞可在多种因素的刺激下发生活化成熟及产生效应,参与肿瘤的免疫应答.长链非编码RNA(long non-codingRNA)是一类多效的调控因子,可参与机...     

肿瘤浸润淋巴细胞输注预防乳腺癌复发

目的　探讨肿瘤浸润淋巴细胞输注预防乳腺癌复发的疗效。方法　从乳腺癌组织及淋巴结中提纯 TIL细胞 ,将其扩增达到一定数量 (>1.0× 10 9个 )后输入乳腺癌患者 ,并进行外周血中的 T细胞亚群及 TIL杀伤活性测定。结果　输注 TIL细胞后 ,机体细胞免疫功能增强 ,TIL最高杀伤率达 78.0 % ,2 6例输注者无 1例复发。结论　乳腺癌患者自身 TIL细胞输注 ,对预防乳腺癌复发有一定的积极作用     

Programmed death ligand 1 expression and tumor infiltrating lymphocytes in neurofibromatosis type 1 and 2 associated tumors

Immune checkpoint inhibitors targeting programmed cell death 1 (PD-1) or its ligand (PD–L1) have been shown to be effective in treating patients with a variety of cancers. Biomarker studies have found positive associations between clinical response rates and PD–L1 expression on tumor cells, as well as the presence of tumor infiltrating lymphocytes (TILs). It is currently unknown whether tumors associated with neurofibromatosis types 1 and 2 (NF1 and NF2) express PD–L1. We performed immunohistochemistry for PD–L1 (clones SP142 and E1L3N), CD3, CD20, CD8, and CD68 in NF1-related tumors (ten dermal and six plexiform neurofibromas) and NF2-related tumors (ten meningiomas and ten schwannomas) using archival formalin-fixed paraffin-embedded tissues. Expression of PD–L1 was considered positive in cases with >?5% membranous staining of tumor cells, in accordance with previously published biomarker studies. PD–L1 expression in tumor cells (using the SP142 and E1L3N clones, respectively) was assessed as positive in plexiform neurofibromas (6/6 and 5/6) dermal neurofibromas (8/10 and 6/10), schwannomas (7/10 and 10/10), and meningiomas (4/10 and 2/10). Sparse to moderate presence of CD68, CD3, or CD8 positive TILs was found in 36 (100%) of tumor specimens. Our findings indicate that adaptive resistance to cell-mediated immunity may play a major role in the tumor immune microenvironment of NF1 and NF2-associated tumors. Expression of PD–L1 on tumor cells and the presence of TILs suggest that these tumors might be responsive to immunotherapy with immune checkpoint inhibitors, which should be explored in clinical trials for NF patients.