食管鳞状细胞癌组织中FGL1的表达、肿瘤浸润淋巴细胞的分布及其意义

目的 探讨食管鳞状细胞癌（ESCC）组织中纤维蛋白原样蛋白1（FGL1）的表达、肿瘤浸润淋巴细胞（TILs）的分布及其与预后的关系。方法 回顾性分析120例ESCC患者临床资料,采用免疫组织化学法检测FGL1的表达,HE镜下评估肿瘤内浸润淋巴细胞（iTILs）和肿瘤间质浸润淋巴细胞（sTILs）的分布情况。生存分析评估患者的预后情况。结果 FGL1在ESCC组织中的阳性率为18.3%（22/120）,其表达与TNM分期、淋巴结转移和TILs有关;ESCC组织中低度iTILs（iTILs≤10%）73例（60.8%）,高度iTILs（iTILs>10%）47例（39.2%）;低度sTILs（sTILs≤10%）82例（68.3%）,高度sTILs（sTILs>10%）38例（31.7%）。iTILs分布与FGL1的表达、肿瘤分化程度和TNM分期有关,sTILs分布与FGL1的表达、肿瘤部位和TNM分期有关。Kaplan-Meier生存分析结果显示肿瘤直径、TNM分期、淋巴结转移、FGL1的表达、TILs的分布与患者预后有关（P<0.05）,多因素Cox回归结果显示,FGL1的表达、TILs的分布和TNM分期是患者预后的影响因素。结论 FGL1的表达与ESCC的不良预后相关,可能成为ESCC的预后生物标志物。FGL1联合TILs可作为预测ESCC预后的生物标志物。     

Immunoexpression of Tumor Infiltrating Lymphocytes(TILS) CD4 + and CD8 + in Oral Squamous Cell Carcinoma (OSCC) in Correlations with Clinicopathological Characteristics and Prognosis

Objective: This study aimed to analyze CD4 +and CD8 + TILs in oral squamous cell carcinoma (OSCC) and to correlate it with histologic grade of malignancy and clinicopathologic data. Methods: The sample was composed of 43 archived specimens. Clinical features and histological grade of malignancy were obtained. The infiltrating intensity of CD4 +, CD8 positive cells were assessed by immunohistochemistry. One-way ANOVA was used to study the association between CD4 +, CD8 + and the grade of OSCC. The cut-off values of the proposed diagnostic indices were received from calculating the coordinates of the receiver operating characteristic (ROC) curve. For clinicopathologic data Independent-Samples T test, Pearson Correlation Coefficient, Correlation Coefficient were used clinicopathologic characteristics. Results: CD4 +and CD8 + were observed in all specimens. CD4 + were more frequent in poorly differentiated specimens (74.14) (P= 0.021<0.05). CD8 + were more frequent in well- differentiated  specimens (51.18). None of these correlations were significant (P=0.454>0.05). CD4 +/ CD8 ratio was higher in low-grade specimens (180.28) (P=0.017<0.05). No differences between CD4 +, CD8 +and CD4 +/ CD8 ratio between poorly- differentiated   and moderately- differentiated   groups ROC P value (0.370, 0.248, 0.126) respectively. there is a difference between CD4 +, CD4 +/ CD8ratio between poorly- differentiated   and well- differentiated   groups ROC P value (0.022, 0.341, 0.012) Sensitivity (0.857, 0.882), specificity (0.706, 0.857) respectively. and no differences between CD8 + poorly- differentiated and well- differentiated groups ROC P value (0.341). there is a difference between CD4 + between moderately - differentiated   and well- differentiated groups ROC P value (0.038) Sensitivity (0.368), specificity (0,765). No significant correlation was obtained with clinicopathologic findings of OSCC. Conclusion: CD4 + and CD4 +/ CD8 + ratio are independent prognostic factor in OSCC.     

乳腺癌微环境中T细胞与临床病理因素的相关性

目的:探讨乳腺癌中间质肿瘤浸润淋巴细胞（TILs）包括CD8+TILs、CD4+TILs、三维淋巴结构（TLS）与临床病理因素和分子亚型的相关性。方法:收集我院2015年至2017年乳腺癌标本200例,经常规石蜡包埋制片,判读HE染色切片间质TLS个数,采用免疫组织化学SP法检测200例病例中CD8+TILs和CD4+TILs浸润密度。结果:乳腺癌间质CD8+TILs、CD4+TILs、TLS与SBR分级、病理TNM分期、Ki67、p53呈正相关;CD8+TILs和CD4+TILs与淋巴结转移呈正相关;与患者年龄呈负相关（均P＜0.05）。CD8+TILs和CD4+TILs呈正相关（P＜0.05）。TLS以CD4+TILs为主,分别与CD8+TILs和CD4+TILs呈正相关（均P＜0.05）。在乳腺癌不同分子亚型中CD8+TILs、CD4+TILs和TLS存在显著差异。结论:乳腺癌中浸润的CD8+TILs、CD4+TILs、TLS反映局部肿瘤微环境的免疫状态,三者密切相关。乳腺癌组织中增多的CD4+TILs诱导CD8+TILs产生CD8+Treg细胞参与乳腺癌的免疫抑制,导致乳腺癌局部免疫功能下降,促进乳腺癌进展及转移。结合乳腺癌分子亚型和局部免疫状态的分析将更有利于预测乳腺癌进展的生物学潜能和对分子靶向性疗效和预后的评估。     

负性共刺激分子B7-H4在结直肠癌及其微环境中的表达及临床意义

负性共刺激分子B7-H4是新近发现的B7家族的新成员。为了检测人结直肠癌组织中肿瘤细胞及肿瘤浸润淋巴细胞(tumor-infiltrating lymphocyte,TIL)上B7-H4的表达以及分析巨噬细胞、T淋巴细胞及其亚群的浸润,进而探讨其相关临床意义,本课题采用免疫组化SP法检测98例人结直肠癌组织中肿瘤细胞及浸润淋巴细胞上B7-H4的表达以及巨噬细胞、T细胞及其亚群的浸润,统计学分析B7-H4的表达与患者临床病理参数、预后、肿瘤相关巨噬细胞(tumor-associated mac-rophage,TAM)以及CD3+T细胞、CD8+T细胞浸润程度的相关性。结果显示,结直肠癌组织中高表达B7-H4分子,而正常组织中基本不表达,并且在肿瘤细胞及浸润淋巴细胞中均检测到B7-H4的表达。统计学分析表明,浸润淋巴细胞中B7-H4的表达与患者年龄、有无淋巴结转移、是否为粘液腺癌有关(P<0.05),与患者总体生存率呈负相关(P<0.05),且B7-H4高表达组织较B7-H4低表达组织CD3+T细胞高度浸润例数显著增加(P<0.05);肠癌细胞中B7-H4的表达与患者肿瘤分期、淋巴结转移、有无远处转移以及Dukes分期有关(P<0.05)。综上所述,人结直肠癌患者的肠癌细胞及浸润淋巴细胞较正常细胞均异常表达B7-H4分子,CD3+T细胞的浸润程度与浸润淋巴细胞中B7-H4的表达呈显著相关而与肿瘤细胞上B7-H4的表达无关,而肿瘤细胞表达B7-H4与肿瘤分期、淋巴结转移、有无远处转移以及Dukes分期有关。上述结果提示肿瘤细胞表达B7-H4与浸润淋巴细胞表达B7-H4可能分别以不同机制参与了肿瘤的发生和进展,进一步分析肿瘤组织中B7-H4在不同细胞表达的生物学和临床意义具有重要的价值。     

Cetuximab strongly enhances immune cell infiltration into liver metastatic sites in colorectal cancer

Cetuximab has activity against colorectal cancers. Recent studies demonstrated that cetuximab induces antibody‐dependent cell‐mediated cytotoxicity via immune cells, and a new immune‐related mechanism of inducing immunogenic cell death. This study aimed to evaluate the immune responses induced by cetuximab in tumor microenvironments at liver metastasis sites of metastatic colorectal cancer patients. We assessed immune cell infiltration in the liver metastatic sites of 53 colorectal cancer patients. These patients were divided into three groups according to the treatment before operation: chemotherapy with cetuximab, chemotherapy without cetuximab, and no chemotherapy. The inflammatory cells in the liver metastatic sites were assessed by hematoxylin–eosin staining, focusing on the invasive margin. The overall inflammatory reaction and number of lymphoid cells were assessed with a four‐point scoring system. We then assessed immune cell infiltration (CD3, CD8 and CD56) in 15 liver metastatic sites. Hematoxylin–eosin staining demonstrated more inflammatory cells in the chemotherapy with cetuximab group than in the other groups (P < 0.001). Of note, inflammatory cells were found in intratumoral areas, and the destruction of cancer cell foci was observed in the chemotherapy with cetuximab group. Moreover, a higher infiltration of CD3+ (P = 0.003), CD8+ (P = 0.003) and CD56+ (P = 0.001) cells was observed in the chemotherapy with cetuximab group than in the other groups. These results suggest that cetuximab might have an immune‐enhancing effect. As such, the immune‐related mechanism of action of cetuximab may enhance the efficacy of combination therapy, such as chemotherapy and immunotherapy using therapeutic peptides.     

Locally advanced breast cancer: Tumor-infiltrating lymphocytes as a predictive factor of response to neoadjuvant chemotherapy

ObjectiveTo evaluate the pathologic response after neoadjuvant chemotherapy in patients with breast cancer according to the stromal tumor-infiltrating lymphocytes (TILs) as well as the evaluation of overall and disease-free survival according to TILs.MethodsA six years (2008–2013) review was done including patients with locally advanced breast cancer that received neoadjuvant therapy and then surgery. An evaluation of the percentage of TILs was done in the pretreatment biopsies and a correlation analysis and survival curves were done.Results187 patients were evaluated. The pathological complete response (pCR) in patients with TILs ≥30% was 58.5%, and in patients with TILs < 30% was 11% (p < 0.001). An Odds Ratio of 8.85 was obtained in patients with TILs ≥30% to achieve a pCR. This relationship was seen in patients with HER2-enriched and triple-negative subtypes. No correlation between TILs and survival was obtained (OS: log-rank; p = 0.834; DFS: log-rank; p = 0.937).ConclusionsThe study of TILs is important because they represent an additional tool to predict the response to neoadjuvant treatment mostly in HER2-enriched and triple-negative subtypes of breast cancer.     

Comprehensive immune characterization and T‐cell receptor repertoire heterogeneity of retroperitoneal liposarcoma

Retroperitoneal liposarcoma (RLPS) is one of the most common subtypes of retroperitoneal soft tissue sarcomas and lacks effective treatment. This study aimed to provide a thorough profile of immune characteristics of RLPS. This study included 56 RLPS patients. Multisite tumor tissues were collected from 16 patients. Immunohistochemistry was carried out to identify CD4⁺, CD8⁺, FoxP3⁺, CD20⁺, or programmed cell death‐1 (PD‐1)⁺ tumor infiltrating lymphocytes (TILs) and  Programmed cell death ligand‐1 (PD‐L1) expression in tumor tissues. Ultradeep sequencing of T‐cell receptor (TCR) β‐chain gene was carried out in 42 tumor samples as well as peripheral blood samples collected from 6 patients. In RLPS, TILs were distributed in 3 patterns and T cells were more prevalent than B cells. Generally, the proportion of TILs decreased and PD‐L1 expression increased with tumor progression. Patients with higher PD‐1/PD‐L1 expression tended to have poorer prognosis, whereas patients with tertiary lymphoid structure tended to have a favorable disease‐free survival. Although T‐cell clones in tumors were quite different from those in peripheral blood, TCR sequencing showed low TCR repertoire reads as well as polyclonal status within tumors, which indicated limited T cell response in the tumors. Both TILs distribution and TCR repertoires suggested spatial immune heterogeneity in RLPS. Our research described the immune landscape of RLPS, and suggested RLPS might be a kind of tumor with low T cell infiltration as well as great immune heterogeneity. Therefore, strategies that can facilitate lymphocytic infiltration and immune reactivity need to be developed in the future to improve the efficacy of immunotherapy.     

The prognostic values of tumor-infiltrating neutrophils,lymphocytes and neutrophil/lymphocyte rates in bladder urothelial cancer

Tumor-infiltrating neutrophils (TINs) and lymphocytes (TILs) are found to play essential roles in many tumors and associate with the prognosis of patients. But, the prognostic values of TINs, TILs and NLR (neutrophils-lymphocytes ratio) in bladder cancer (BC) are still undefined. The object of our study was to systematically interrogate the associations of these immune cells with clinical outcomes of BC patients. In our study, a total of 102 patients pathologically diagnosed with BC were included. CD66b⁺ and CD8⁺ antibodies were used to mark neutrophils and CD8⁺ lymphocytes by immunohistochemistry. The results found that TINs and NLR were significantly associated with pathological T-stages of tumors (P?<?0.01), but TILs were not. And TINs were also related to pathological tumor grades (P?=?0.012). Regarding the prognostic values, TINs was related to the high risk of recurrence in non-muscle invasive BC (NMIBC) patients. Elevated TINs and NLR were associated with poor overall survivals of BC patients, whereas higher TILs were related to longer survivals (P?<?0.01). Multivariate analysis showed that both of TINs (HR 2.427, 1.024-5.752, P?=?0.044) and NLR (HR 3.529, 1.147-10.864, P?=?0.028) were independent unfavorable prognosis markers. In conclusion, Tumor infiltrating immune cells, including TINs, TILs and NLR were important markers in predicting the prognosis of bladder cancer patients. TINs and NLR were more likely to be negative predictors, but TILs were favorable in patients with BC.     

人骨转移瘤肿瘤浸润淋巴细胞的过继免疫治疗

肿瘤浸润淋巴细胞(TIL)的杀伤肿瘤细胞效应比淋巴因子活化杀伤细胞(LAK)更为有效低毒。作者用手术与活检的方法从骨转移瘤标本中分离TIL,经体外重组白细胞介素—2(rIL—2)激活并扩增,过继转输10例骨转移瘤病例,完全应答1例,部分应答2例,弱应答3例,无应答4例。有效应答30%。