Prognostic value of programmed death.1, programmed death-ligand 1, programmed death-ligand 2 expression, and CD8(+) T cell density in primary tumors and metastatic lymph nodes from patients with stage T1.4N+M0 gastric adenocarcinoma

Background: Anti-programmed death-1/programmed death-ligand 1(PD-1/PD-L1) immunotherapy has been proved to be effective on gastric cancer in ongoing clinical trials. However, the value of PD-L1 in predicting responses of patients with gastric cancer to anti-PD-1/PD-L1 immunotherapy is controversial. Some studies suggested that intra-and inter-tumoral heterogeneity of PD-L1 expression might explain the controversy.This study aimed to analyze the expression of PD-L1, PD-L2, and PD-1 as well as CD8(+) T-cell density in primary tumors and lymph nodes from patients with stage T1-4 N+M0 gastric adenocarcinoma to explore the heterogeneity of PD-1 signaling pathway molecules.Methods: In primary tumors and metastatic as well as non-metastatic lymph nodes from patients with stage T1-4 N+M0 gastric adenocarcinoma, we detected PD-L1 and PD-L2 expression with immunohistochemistry. CD8(+)T-cell density in primary tumors and PD-1 expression on CD8(+)T cells were detected with immunofluorescence. Univariate analysis was used to determine the prognostic values of them. Cox proportional hazard regression model was used to identify independent risk factors that affect patients' overall survival and disease-free survival.Results: Among 119 eligible patients who had undergone surgical resection, the positive rate of PD-L1 was higher in metastatic lymph nodes than in primary tumors(45.4% vs. 38.7%, P = 0.005); the positive rate of PD-1 on CD8(+)T cells was significantly higher in primary tumors and metastatic lymph nodes than in tumor-free lymph nodes(both P < 0.001). The intensity of PD-1 expression on CD8(+) T cells in primary tumors and in metastatic lymph nodes were stronger than that in tumor-free lymph nodes from the same patient. Beside, the positive rate of PD-L2 did not show any differences between primary tumors and metastatic lymph nodes. In multivariate analysis, PD-L1 expression,PD-L2 expression, a low density of CD8(+) T cells in primary tumors, and PD-1 expression on CD8(+) T cells in primary tumors were associated with poor prognosis.Conclusion: The expression of PD-L1 is heterogeneous in primary tumors and in metastatic lymph nodes from patients with stageT1-4 N+M0 gastric adenocarcinoma, which might explain the inconsistent results in assessing the prognostic value of PD-L1 expression in previous studies.

Stromal CD4+ and CD8+ T Cells in Human Breast Carcinomas. Its Correlation with Chemokine MIG/CXCL9

The interaction between some chemokines with tumoral and stromal cells can influence tumor progression. CXCL9, a monokine induced by interferon gamma (MIG), targets lymphocytes.The aim of our study was to identify and quantify CD4+ and CD8+ T cells in the stroma of human breast cancer and correlate them with the presence of MIG/CXCL9. In 58 specimens of human breast carcinoma and 10 normal breast tissue from mammoplasty surgery, immunohistochemistry and ELISA methods were performed. The number of CD4+ and CD8+ T cells in breast cancer tissue was significantly increased compared with normal breast tissue with a clear predominance of CD8+ T cells. MIG/CXCL9 levels were significantly elevated respect normal breast tissue. This chemokine correlated with the number of CD8+ T cells only in non-metastatic tumors. These data suggest that MIG targets cytotoxic T cells. Their recruitment into breast carcinoma can play a critical role in malignant progression, inhibiting the production of metastasis.

T-regulatory lymphocytes in peripheral blood of gastric and colorectal cancer patients

AIM: To assess the absolute number of T-regulatory cells (Tregs; CD4+CD25+Foxp3+) in the peripheral blood of gastric and colorectal cancer patients. METHODS: We enrolled 70 cancer patients (33 gastric cancer, 37 colorectal cancer) and 17 healthy volunteers. The CD3+CD4+ lymphocytes and CD4+CD25+Foxp3+ Tregs in the peripheral blood were analyzed with flow cytometry. The absolute numbers of Tregs were calculated based on the CD4+CD25+Foxp3+ cells percent-age of CD3+CD4+ cells and the absolute numbers of CD3+CD4+ cells per microliter. RESULTS: The mean number of CD4+CD25+Foxp3+ cells per microliter in colorectal cancer patients was 15.7 (SD: 21.8), for gastric cancer patients 12.2 (SD: 14.3), and for controls 17.5 (SD: 11.4). The absolute number of Tregs was significantly lower in gastric cancer patients than in controls (P = 0.026). There was no statistically significant difference for gastric vs colorectal cancer or colorectal cancer vs controls. The absolute number of Tregs was also significantly depressed in N+ vs Ncancer patients [22.0 (27.7) vs 10.1 (9.0), P = 0.013], and in the subgroup of gastric cancer patients [30.3 (27.6) vs 9.6 (8.0), P = 0.003]. No statistical difference was observed in the proportion of Tregs in the CD4+ population between the groups. CONCLUSION: The absolute number of Tregs in peripheral blood of gastric cancer but not colorectal cancer patients was significantly decreased in comparison with that in healthy controls.

The Hematopoietic and Immunomodulatory Effect of rhIL-12 for Liver Cancer

Purpose: To explore the effect of rhIL-12 on the number of the blood cells and CD4/8+ T, CD45+ leukocytes, and CD56+ NK cells in liver cancer patients following radiation therapy. Methods: We selected forty liver cancer patients who carried out by cyber knife (the patients were given 5 Gy every time for 5 times continuously) to observe the size of the tumor. After thirty hours, rhIL-12 was injected into the liver cancer patients via subcutaneous at the concentration of 50 ng/kg, 100 ng/kg, 200 ng/kg and 300 ng/kg in different patients, respectively. And there were ten patients in the four groups, respectively. The twenty patients who were selected from the hospital without rhIL-12 treatment were used as controls. All the blood cells were collected from different groups on day 0, hour 12, day 7, day 14, day 21 and day 28 after rhIL-12 treatment, respectively. The full number of blood cells in every group was analyzed by ELISA. The number of CD4/8+ T, CD45+ leukocytes, and CD56+ NK cells were detected by Flow Cytometry. After one month with rhIL-12 treatment, ECOG and WHO were used to evaluate the prognosis of liver cancer. Results: In present study, we found that the number of blood cells was significantly decreased on day 0 - day 3, while recovered from day 7 - day 14 and down-regulated on day 21 after rhIL-12 treatment. The number of CD4/8+ T, CD45+ leukocytes, and CD56+ NK cells was elevated with any concentration of rhIL-12. Furthermore, results showed that number of white blood cells was obviously higher than in patients without rhIL-12 treatment (P < 0.05). However, there was no significant difference of erythrocyte and platelet, between groups treated with rhIL-12 and control groups. In addition, the immune cells including CD4/8+ T, CD45+ leukocytes, and CD56+ NK cells were reduced on day 0 - day 3, recovered from day 7, and then decreased from day 21 in rhIL-12 treatment groups related to control groups (P < 0.05). Furthermore, studies showed that five patients developed symptoms of fever, bilirubin increased and liver dysfunction with the dose of 300 ng/kg. So we found that the safe and well-tolerated human dose of 200 ng/kg is within this efficacious range based on exposure parameters through the research. Higher ECOG and WHO scores were observed in rhIL-12 treatment groups compared to control groups (P = 0.025, P = 0.044, respectively). Conclusion: Our results suggested that rhIL-12 could recover the liver cancer induced aberrant blood cell number and CD4/8+ T, CD45+ leukocytes, and CD56+ NK cells , which may be an effective method to alleviate the progress of liver cancer and played an important role in treating liver cancer.

肿瘤浸润淋巴细胞和树突细胞对胃癌患者预后的影响

目的研究胃癌组织中CD4+T、CD8+T、CD83+DC肿瘤浸润细胞的临床意义及预后影响。方法采用免疫组化SP法检测120例胃癌组织及其癌旁正常组织中CD4T、CD8T、CD83+DC表达情况。结果 120例胃癌组织中CD4、CD8、CD83DC表达的阳性率分别为52.4%、63.7%、46.3%,均低于相应的癌旁正常组织(P<0.05)。胃癌中CD4+T、CD8+T、CD83+DC的5年生存率分别为56.5%、58.1%、48.4%,高于CD4+T、CD8+T、CD83-DC组的5年生存率,差别无显著性意义(P>0.05)。标本分成CD4/8(+/+)CD83+DC、CD4/8(+/-)CD83+DC、CD4/8(-/+)CD83+DC、CD4/8(-/-)CD83+DC、CD4/8(+/+)CD83-DC、CD4/8(+/-)CD83-DC、CD4/8(-/+)CD83-、CD4/8(-/-)CD83-DC八组,其中CD4/8(+/+)CD83+DC组的5年生存率为68.7%,高于其余组(P<0.05)。胃癌中CD4/8(+/+)与CD83+DC之间有相关性(相关系数r=0.72,P<0.01)。结论胃癌组织中CD4/8(+/+)CD83+DC表达可能有利于胃癌患者术后的预后,并且CD4/8(+/+)和CD83+DC有密切关系。

维吾尔族妇女局部晚期宫颈鳞癌外周血T细胞亚群与临床特征和预后关系

目的观察新疆地区维吾尔族局部晚期宫颈鳞癌患者外周血T细胞亚群与临床特征、预后的关系。方法选择2015-2018年间新疆医科大学附属肿瘤医院经组织病理证实的ⅡB-ⅣA期宫颈鳞癌患者185例,回顾性分析外周静脉血T细胞亚群与临床特征及预后的关系。结果CD^+4T细胞、CD^+8T细胞、CD^+4/CD^+8T细胞比值与临床分期、肿瘤最长径、体质量指数(BMI)有关(均P<0.05)。国际妇产科协会(FIGO)分期越晚、最长径越大、BMI越高,CD^+8T细胞越高,CD^+4T细胞、CD^+4/CD^+8T细胞比值越低;淋巴结转移者,CD^+4T细胞减低。单因素分析结果显示FIGO分期、年龄、淋巴结转移、肿瘤最长径、BMI、CD^+4T细胞、CD^+8T细胞、CD^+4/CD^+8T细胞比值及治疗方式是影响局部晚期宫颈鳞癌患者总生存(OS)的重要因素;多因素分析结果显示BMI、治疗方式、CD^+4T细胞及肿瘤最长径是影响患者OS的独立预后因素(均P<0.05)。结论维吾尔族宫颈癌患者外周血T细胞亚群水平失调,CD^+4T细胞、CD^+8T细胞、CD^+4/CD^+8T细胞比值与宫颈癌FIGO分期、肿瘤最长径、BMI有关,CD^+4T细胞还与淋巴结转移相关。BMI、治疗方式、肿瘤最长径及CD^+4T细胞是影响患者OS的独立预后因素。

大麻素受体2与子宫颈癌组织内浸润T细胞亚群的关系

目的研究大麻素受体2(cannabinoid receptor 2,CB 2)在人子宫颈癌组织中的表达及其与肿瘤内浸润T细胞亚群,包括FOXP 3+T、CD 4+T、CD 8+T细胞的关系。方法选取2016年德州市中医院病理科保存的人体手术切除标本90例,纳入30例正常人宫颈组织为对照A组、30例人高级别宫颈上皮内瘤变(cervical intraepithelial neoplasia,CIN)Ⅱ~Ⅲ级组织为对照B组、30例人子宫颈癌组织为观察组。采用免疫组织化学法及实时定量聚合酶链反应(quantitative real-time polymerase chain reaction,RT-PCR)检测CB 2在各组中表达情况,采用间接免疫荧光双标法检测观察组组织内浸润FOXP 3+、CD 4+T、CD 8+T细胞数目。结果 CB 2蛋白及mRNA在3组中均有表达,且在观察组中表达最高、对照B组中次之、对照A组中表达量最低,各组间差异均有统计学意义(P<0.05);观察组CB 2蛋白及mRNA的表达均与组织内浸润的CD8+T呈负相关(P<0.05)。结论 CB 2在人子宫颈癌组织内存在高表达现象,且与组织内浸润CD 8+T细胞数量减少有一定相关性。

转染肿瘤mRNA的树突状细胞疫苗诱导抗肝癌免疫研究

目的 探讨转染原发性肝癌(HCC)mRNA的树突状细胞(DC)能否诱导抗肿瘤特异性细胞毒性T淋巴细胞(CTL)。方法 采用HCC患者外周血单核细胞(PBMC)体外刺激分化为DC细胞;从人肝癌HepG 2细胞和3例HCC患者的肝癌组织中体外扩增mRNA。以mRNA转染DC细胞,并与PBMC混合培养诱导扩增CTL。流式细胞计数仪检测培养细胞中CD3 +、CD4+、CD8+细胞的比例。51Cr释放法测定CTL的杀瘤活性。结果 经扩增人肝癌HepG 2mRNA和2例AFP(+ )患者的AFP(+ )HCCmRNA诱导3周后,CD3 +、CD8+细胞占淋巴细胞总数由诱导前的2 7.8%、2 6.5 %、2 9.6%升高至89.3 %、73 .6%、86.8% ;而经扩增AFP(-)HCCmRNA诱导3周后,CD3 +、CD8+细胞占淋巴细胞总数由诱导前的2 5 .4%升高至5 3 .6%。转染HepG 2细胞和AFP(+ )的患者HCCmRNA的DC诱导的CTL对HepG 2细胞杀瘤活性明显高于AFP(-)的患者,其杀瘤特性由MHC I限制的CD8+T细胞所介导。结论 HCCmRNA体外转染DC能诱导肿瘤特异性CTL 。