金米冲剂对35例肝癌病灶免疫状况影响

[目的]探讨金米冲剂(Star-Ⅰ)对原发性肝癌患者病灶局部免疫状况的影响.[方法]35例原发性肝癌患者分别在用药前后,在超声引导下经皮肝脏穿刺活检各1次,取出的组织标本用特异性抗体免疫组织化学染色,检测病灶区域浸润的T淋巴细胞、自然杀伤细胞、巨噬细胞阳性率.[结果]金米冲剂对原发性肝癌患者局部浸润免疫细胞的功能有提高(P＜0.05),这些浸润免疫细胞主要为T细胞、自然杀伤细胞和巨噬细胞.[结论]金米冲剂对肝癌患者病灶局部细胞免疫功能有提高.     

肝细胞癌患者病灶局部Treg细胞分析及其临床意义

[目的]探讨肝细胞癌患者病灶局部Treg细胞的变化规律及其临床意义。[方法]收集40例原发性肝细胞型肝癌(HCC)患者经手术切除的肝癌组织及非癌肝组织(距肿瘤边缘>5cm)标本,用流式细胞仪分别检测肝癌组织、非癌肝组织中调节性T淋巴细胞(Treg细胞)的数量及变化规律,并且分析Treg细胞表达数量与预后的关系。[结果]肝癌病灶局部有不同程度免疫细胞浸润,主要为T淋巴细胞,其中Treg淋巴细胞含量高于非肿瘤组织。Ⅲ、Ⅳ期患者Treg细胞含量(7.7%±4.03%)明显高于Ⅰ、Ⅱ期患者(3.7%±1.6%)(P=0.037)。高Treg细胞含量组(≥6%)患者的预后明显差于低Treg细胞含量组(<6%)(P=0.005)。[结论]肝癌组织中,对免疫起抑制作用的Treg细胞比例增加。Treg细胞含量高与预后差相关。     

原发性肝细胞癌组织中T、B淋巴细胞浸润的研究

目的 研究原发性肝细胞癌(HCC)组织中T、B淋巴细胞的浸润情况,探讨肿瘤的局部免疫功能状态.方法 用免疫组织化学S-P法对50例HCC和10例正常肝组织进行CD3、CD20染色,对阳性细胞数进行定量分析.结果 T、B淋巴细胞的数量在HCC癌组织及癌旁肝组织高于正常肝组织(P＜0.05),癌旁有肝硬化组织高于无肝硬化组织(P＜0.05);T淋巴细胞的数量在癌组织、癌旁肝硬化组织高于B淋巴细胞(P＜0.05);T或B淋巴细胞数量与HCC分化程度、临床分期及有否门静脉癌栓形成无关(P＞0.05).结论 HCC局部以细胞免疫为主;淋巴细胞计数不能作为判断患者预后的指标.     

原发性肝细胞癌组织免疫微环境的分析

目的： 探讨原发性肝细胞癌（hepatocellular carcinoma,HCC）组织中免疫细胞的浸润及分布情况。 方法： 采用免疫组化S-P法,检测采集自福建省肿瘤医院30例HCC癌组织、癌周组织（距肿瘤边缘<2 cm）、非癌肝组织（距肿瘤边缘＞5 cm）石蜡标本中CD3、CD4、CD8、FoxP3、CD20、CD56、CD68的表达。 结果： 免疫组织化学结果显示,HCC患者CD3+、CD4+、FoxP3+细胞数在癌周组织中最高,癌组织中其次,非癌肝组织中最低（P<0.05）;CD8+、CD56+、CD68+细胞数在癌周组织中最高,非癌肝组织其次,癌组织中最低（P<005）;CD20+细胞数在肝癌组织、癌周组织、非癌肝组织中无明显差异（P>005）。肝癌组织中CD3+、CD4+、CD8+、CD56+、CD68+细胞主要分布于癌巢中的间质区,FoxP3+细胞呈散在分布。 结论: 肝癌组织局部微环境中,杀伤性免疫细胞减少,抑制性免疫细胞增加,从而导致局部免疫抑制状态。     

胃癌微环境中免疫活性细胞与患者预后的关系

目的:主要研究胃癌局部浸润的记忆性T 细胞、树突状细胞和细胞毒性T 淋巴细胞与患者预后的关系。方法:选取60例胃癌组织蜡块,用免疫组化方法检测胃癌局部浸润的记忆性T 细胞、树突状细胞和细胞毒性T 淋巴细胞的数量及分布,分析局部免疫活性细胞与胃癌患者临床病理特征及预后的关系。结果:胃癌局部免疫活性细胞高表达者,淋巴结转移率较低。单因素分析显示,淋巴结转移、肿瘤直径、TNM分期、肿瘤组织局部浸润的记忆性T 细胞、细胞毒性T 淋巴细胞和树突状细胞及免疫细胞联合表达是胃癌患者预后的影响因素。多因素分析显示,淋巴结转移和记忆性T 细胞是胃癌患者预后的独立影响因素。结论:胃癌局部浸润的免疫活性细胞可以很好地预示淋巴结的转移情况。胃癌局部浸润的记忆性T 细胞对判断患者的预后有一定意义。      

肝细胞性肝癌组织浸润淋巴细胞表型与分布研究

目的:肝细胞性肝癌组织浸润淋巴细胞与外周血T 细胞表型可能与肿瘤进展及预后相关,本研究检测肝癌患者组织及外周血T 细胞表型与分布,分析淋巴细胞表型变化与预后的关系。方法:分析2007年10月至12月中山医院147 例肝癌及癌旁组织浸润淋巴细胞表型（T 细胞或B 细胞表面标志物:CD3、CD8、CD4、CD20、CD19、Foxp 3）,表型与临床病理特征及预后的关系;检测26例肝癌外周血CD3、CD8、CD4 +T细胞数量并其比例变化。结果:癌巢内肿瘤浸润细胞明显少于癌周组织（P < 0.01）,癌周淋巴细胞主要分布于癌旁正常肝组织、汇管区,其与患者肝炎病史及肝硬化相关,表型以CD3 +T细胞为主,其中又以CD8 + 细胞毒性T 细胞为主;CD4 染色在多数病例为阴性,Foxp 3 仅在个别病例（15/ 109）呈阳性。肿瘤浸润淋巴细胞B 细胞标志CD20、CD19均为阴性。肿瘤组织内CD8 +T细胞浸润数量与预后正相关,而癌周浸润淋巴细胞数目与患者转移及复发无显著关系。结论:肝癌肿瘤浸润细胞在癌巢内明显少于癌周组织,肿瘤及癌周浸润细胞以CD8 + 细胞毒性T 细胞为主。肿瘤组织内CD8 +T细胞浸润数量与预后相关,而癌周浸润淋巴细胞数量与患者转移及复发无显著关系。      

T细胞杀伤肝癌的机制及其阻断激活T细胞凋亡的研究

研究肝癌组织中是否存在激活T细胞以及如何阻断激活T细胞凋亡,增加激活T细胞寿命对于肝癌的治疗具有重要意义.我们应用免疫组化及原位杂交技术研究20例肝癌组织中肿瘤侵润淋巴细胞(TIL)的穿孔素及Fas-L表达情况,结果表明80%肝部癌组织中存在着表达穿孔素及Fas-L的TIL细胞,表明绝大多数肝癌组织中存在着激活T细胞,为体内、外扩增这种具有杀伤活性的激活T细胞提供实验基础.其中No.14肝癌组织中可见大量TIL细胞侵润,绝大多数TIL细胞表达穿孔素及Fas-L,该例病人已1年6个月无复发,提示大量激活T细胞存在可能有利于肿瘤治疗,然而除该例病人外,其余病人的TIL的穿孔素及Fas-L阳性率<10%,表明仅有极少量TIL细胞处于免疫激活状态,具有杀     

微波治疗肝癌后局部细胞免疫变化及其对临床疗效影响

背景与目的：机体免疫细胞可识别并排斥肿瘤细胞，在临床微波凝固治疗肝癌后免疫细胞的局部浸润的变化和细胞免疫对临床疗效的影响值得探讨。本研究分析超声引导下微波凝固治疗肝癌局部免疫细胞浸润变化，并探讨了局部细胞免疫对临床疗效的影响。方法：超声引导微波凝固治疗原发性肝癌患者89例，于微波治疗前后进行肝穿组织活检，经免疫组化染色后比较治疗前后局部CD4^＋、CD57^＋和CD68^＋浸润变化，并观察肝癌复发情况，分析局部免疫细胞浸润程度在复发和未复发组间是否存在差异。结果：微波治疗后CD4^＋、CD57^＋和CD68^＋局部浸润程度较治疗前显著增加（P〈0．05）；微波术前，CD4^＋和CD57^＋在未复发组阳性细胞密度显著高于复发组（P〈0．05）。结论：微波凝固治疗肝癌后局部免疫细胞浸润增加，且局部免疫细胞的浸润程度对其临床疗效可有一定的影响。     

肺癌组织中免疫活性细胞和β2微球蛋白的免疫组化研究

应用单克隆抗体和免疫组化ABC法对53例肺癌的局部免疫状态及β_2微球蛋白(简称β_2 M)表达进行研究。结果表明:①机体对肺癌组织以细胞免疫为主,局部反应主要为T细胞,伴少量巨噬细胞和B细胞。免疫细胞主要分布于肺癌间质,仅少数散在于癌巢中。②肺癌局部T细胞多处于非活化状态。OKT_4/OKT_8比值各例不同,平均为1.24±0.59。③有淋巴结转移的肺癌组,其局部T细胞和活化T细胞数低于无转移组,而巨噬细胞则是转移组高于无转移组。鳞癌中活化T细胞和巨噬细胞高于未分化癌。以上差别均有显著或极显著意义。④肺癌分化好者β_2 M表达强,分化差者表达弱。β_2 M的表达与全T,辅助T和活化T细胞的浸润呈正相关。本文认为肺癌组织中免疫细胞的浸润及其活性状态可作估计预后的指标之一。β_2 M表达可做肺癌分级的参考指标。     

浸润T淋巴细胞与食管癌生物学特性关系的研究

本文以ANAE标记法对40例食管癌组织中浸润的T淋巴细胞及其亚群进行定量组织学研究,观察T淋巴细胞浸润程度与食管癌生物学特性之间的关系。结果表明,癌组织中T淋巴细胞浸润密度随癌组织分化程度的降低而减少,癌组织中浸润的T淋巴细胞与癌组织的浸润和转移有密切关系。癌组织浸润深度深和有局部淋巴结转移者,T淋巴细胞浸润数量少,反之则较多 在分化差和有淋巴结转移的癌组织中,T淋巴细胞的减少主要为D~+型细胞减少,S~+型细胞相对增加,D~+/S~+比值下降     

Associations Between Infiltrating Lymphocyte Subsets and Hepatocellular Carcinoma

Aims: We aimed to analyze the phenotype of tumor-infiltrating lymphocytes (TILs) and non-tumor infiltrating lymphocytes (NILs) in HCC and non-tumor tissues, and evaluate relationships between changes in these cells and the prognosis of HCC. Methods: Lymphocytes were isolated from HCC and corresponding non-tumor tissues and tested by flow cytometry. For comparison, clinical parameters were analyzed. Results: Compared with the non-tumor tissue, tumor tissue had a lower intensity of NK, NKT andCD8+T cell infiltration. TILs had higher intensity of CD4+CD25+Foxp3+regulatory T cell (Treg cells) infiltration compared with that in NILs. The prevalence of Treg cells was associated with fewer CD8 + T lymphocytes in the HCC immune microenvironment. The frequencies of NK cells and CD8+T cells in TILs of HCC patients with metastasis less than 12 months were lower than those without metastasis. However, the frequency of Treg cells was higher than those without metastasis. Conclusion: These results suggest that the frequencies of CD8+T, NK and NKT cells as well as Treg cells in the tumor tissue of HCC are significantly associated with patient survival, and could be applied as predictive indicators for HCC prognosis.     

胃癌细胞凋亡与微环境免疫细胞关系的定量研究

目的探讨胃癌细胞凋亡与其微环境中的免疫细胞凋亡和浸润程度的关系。方法应用免疫组织化学技术标记40例手术切除的胃腺癌组织微环境中的树突状细胞(DC)、T细胞和NK细胞,并对其浸润程度进行定量评价;应用TUNEL技术标记上述组织胃癌细胞与免疫细胞的凋亡细胞,并定量测出凋亡水平,最后分析胃癌细胞凋亡水平与免疫细胞的凋亡程度和不同免疫细胞浸润程度的关系。结果本组胃癌细胞的凋亡指数与其免疫细胞的凋亡指数呈显著负相关(r=-0.868,P<0.001),而与微环境中DC、T细胞和NK细胞的定量浸润水平呈显著正相关(r值分别为0.823、0.764和0.708,P值均<0.001)。结论胃癌细胞的凋亡水平与其微环境中的免疫细胞的浸润程度和凋亡水平密切相关。     

FOXP3+ regulatory T cells affect the development and progression of hepatocarcinogenesis.

PURPOSE: Tumor-infiltrating lymphocytes represent the host immune response to cancer. CD4+CD25+FOXP3+ regulatory T cells (Tregs) suppress the immune reaction. The aim of the present study was to investigate the clinicopathologic significance and roles of Tregs and CD8+ T cells during hepatocarcinogenesis. EXPERIMENTAL DESIGN: We examined the infiltration of FOXP3+ Tregs and CD8+ T cells in the tumor stroma and nontumorous liver parenchyma using 323 hepatic nodules including precursor lesions, early hepatocellular carcinoma (HCC), and advanced HCC, along with 39 intrahepatic cholangiocarcinomas and 59 metastatic liver adenocarcinomas. We did immunohistochemical comparative studies. RESULTS: The prevalence of Tregs was significantly higher in HCC than in the nontumorous liver (P<0.001). The patient group with a high prevalence of Tregs infiltrating HCC showed a significantly lower survival rate (P=0.007). Multivariate analysis revealed that the prevalence of Tregs infiltrating HCC was an independent prognostic factor. The prevalence of Tregs increased in a stepwise manner (P<0.001) and that of CD8+ T cells decreased during the progression of hepatocarcinogenesis (P<0.001). Regardless of the presence of hepatitis virus infection or histopathologic evidence of hepatitis, the prevalence of Tregs was significantly increased in nontumorous liver bearing primary hepatic tumors. CONCLUSIONS: Tregs play a role in controlling the immune response to HCC during the progression of hepatocarcinogenesis. It has been suggested that primary hepatic cancers develop in liver that is immunosuppressed by a marked infiltration of Tregs. A high prevalence of Tregs infiltrating HCC is thought to be an unfavorable prognostic indicator.     

Assessment of cytotoxic T-lymphocyte phenotype using the specific markers granzyme B and TIA-1 in cervical neoplastic lesions.

Cervical carcinomas are closely associated with high-risk human papillomavirus (HPV) types and are preceded by cervical intraepithelial neoplasia (CIN). Most CIN lesions regress spontaneously and will not evolve to invasive carcinoma. The cellular immune system mediated by cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells are thought to play an important role in the ultimate decline of CIN lesions. Although TIA-1 is constitutively expressed in the majority of circulating T cells and defines a subpopulation of CD8+ T cells with cytotoxic potential, granzyme B is only expressed in CTLs upon activation. In the present study we have evaluated the expression of these proteins by lymphocytes present in 24 randomly chosen CIN lesions with increasing degree of atypia and in 14 cervical squamous cell carcinomas. As major histocompatibility complex (MHC) class I expression is frequently down-regulated in HPV-induced lesions, thus possibly frustrating tumour cell recognition by infiltrating CTLs, these lesions were also analysed for MHC class I expression. The results indicated that in most CIN lesions only a minority of CTLs are activated, whereas in some carcinomas a massive infiltration of activated, i.e. granzyme B-positive, CTLs were observed. The percentage of activated CTLs was not related to expression of MHC class I on neoplastic cells. These results suggest that in some carcinomas proper activation of CTLs occurs but that most likely local factors or immunoselection of resistant neoplastic cells inhibit a proper response of CTLs to these neoplastic cells.     

Intranodal antitumor immunocyte infiltration in node-negative gastric cancers.

The status and role of immunocytes and dendritic cells in regional lymph nodes in patients with gastric cancer are examined in this study. Forty-nine patients with gastric cancer who underwent curative resection were enrolled in the present study. These patients had no lymph node metastases according to a histological examination. The infiltration of natural killer (NK) cells, dendritic cells, and MIB-1-positive immunocytes was investigated. Based on the Japanese Classification of Gastric Carcinoma, regional lymph nodes were divided into three compartments: (a) compartment 1 (lymph node station numbers 1-6); (b) compartment 2 (lymph node station numbers 7-12); and (c) compartment 3 (lymph node station numbers 14 and 16). Dendritic cells and MIB-1-positive immunocytes infiltrated compartment 1 lymph nodes in increased numbers compared with the lymph nodes of compartments 2 or 3 (P < 0.05). Conversely, intranodal NK cell infiltration did not differ significantly among the three compartments. The incidence of intranodal dendritic and MIB-1-positive cell infiltration in patients with submucosal gastric cancer was significantly higher than in patients with tumors that invaded beyond the muscularis propria. The decreased expression of these immunological markers correlated well with recurrent disease, regardless of tumor depth. The immunocyte level is higher in lymph nodes near the primary tumor (compartment 1) than in those that are distant from the tumor (compartments 2 and 3). This pertains to all three markers, i.e., NK, dendritic, and MIB-1-positive cells. Unlike dendritic and MIB-1-positive cells, intratumoral infiltration of NK cells did not correlate well with either lymph node compartment or the depth of tumor invasion. The degree of NK cell infiltration may be directly associated with antitumor effects, especially in compartment 1. A decrease in all three markers is associated with tumor recurrence.     

肝细胞癌端粒酶活性及hTERT mRNA表达与术后早期复发的关系

目的：研究肝细胞癌端粒酶活性及人端粒酶逆转录酶（hTERT）mRNA表达与肝细胞癌术后早期复发的关系。方法：采用ELISA—TRAP法检测60例肝癌组织及其癌旁组织端粒酶活性，RT—PCR法检测hTERT mRNA表达，5例正常肝脏组织作为对照。分析端粒酶活性及hTERT mRNA表达与临床病理之间的关系。结果：肝癌组织端粒酶活性及hTERT mRNA表达阳性率分别为86．7％（52／60）及90％（54／60），癌旁组织端粒酶活性及hTERT mRNA表达阳性率分别为40％（24／60）及43．3％（26／60）。正常肝脏组织均未检测到端粒酶活性及hTERT mRNA表达。癌旁组织端粒酶活性及hTERT mRNA表达与术后早期复发及包膜浸润、门静脉侵犯、肝内转移等恶性肿瘤的恶性生物学行为有关。结论：癌旁组织端粒酶活性及hTERT mRNA表达可能是肝细胞癌术后早期复发的预后指标。     

Sequential pathological and immunologic analysis of percutaneous microwave coagulation therapy of hepatocellular carcinoma.

PURPOSE: To evaluate sequential pathologic and immunologic changes and their prognostic significance after percutaneous microwave coagulation therapy (PMCT) of hepatocellular carcinoma (HCC). METHODS: Eighty-nine nodules in 82 consecutive patients were studied. The 89 nodules were divided into two groups: a treatment group, with 82 primary nodules (average dimension was 3.4 +/- 1.2 cm) in 82 patients, and a control group, of seven nodules (average dimension was 1.4 +/- 0.6 cm) in seven patients. The criteria for a nodule's inclusion in the control group was that the nodule was one of two nodules in the same patient and that the two nodules were located in different liver lobes. This guarantees that while one nodule is treated by PMCT, the distant one will not be directly affected by the microwave thermal field. The control group nodules were treated after the study was completed. Specimens were taken with ultrasound-guided liver biopsy from the treated nodule and the control nodule, pre- and post-PMCT. Infiltration by T-lymphocytes, B-lymphocytes, NK cells and macrophages in the tumour tissue were observed immunohistochemically using a panel of monoclonal antibodies against CD3, CD45RO, CD20, CD56 and CD68. The extent of immune cell infiltration was compared both before and after PMCT, as well as between the treated and control nodules. The relationship between the prognosis and the extent of immunocyte infiltration was analysed. RESULTS: The patients were followed for 2-26 months (mean 14.6 +/- 6.3) post-treatment. The recurrence rates at 1 and 2 years were 20.4% and 28.1% within the liver in treatment group, respectively. The survival rates at 1 and 2 years were 92.5% and 75.3% for the treatment group. T-cells, NK cells and macrophages increased significantly in both treated and untreated nodules after PMCT, albeit less markedly within untreated nodules when compared to the treated ones. There is a statistically significant correlation between survival outcome and the extent of immunocyte infiltration. CONCLUSIONS: For inoperable HCC patients, PMCT is one of the treatment choices shown to be effective. Apart from its tissue coagulation effect, an increased systemic immune response directed against the tumour may also play an important role in improved survival.     

Sequential pathological and immunologic analysis of percutaneous microwave coagulation therapy of hepatocellular carcinoma

Purpose: To evaluate sequential pathologic and immunologic changes and their prognostic significance after percutaneous microwave coagulation therapy (PMCT) of hepatocellular carcinoma (HCC). Methods: Eighty-nine nodules in 82 consecutive patients were studied. The 89 nodules were divided into two groups: a treatment group, with 82 primary nodules (average dimension was 3.4 &#45 1.2 cm) in 82 patients, and a control group, of seven nodules (average dimension was 1.4 &#45 0.6 cm) in seven patients. The criteria for a nodule's inclusion in the control group was that the nodule was one of two nodules in the same patient and that the two nodules were located in different liver lobes. This guarantees that while one nodule is treated by PMCT, the distant one will not be directly affected by the microwave thermal field. The control group nodules were treated after the study was completed. Specimens were taken with ultrasound-guided liver biopsy from the treated nodule and the control nodule, pre- and post-PMCT. Infiltration by T-lymphocytes, B-lymphocytes, NK cells and macrophages in the tumour tissue were observed immunohistochemically using a panel of monoclonal antibodies against CD3, CD45RO, CD20, CD56 and CD68. The extent of immune cell infiltration was compared both before and after PMCT, as well as between the treated and control nodules. The relationship between the prognosis and the extent of immunocyte infiltration was analysed. Results: The patients were followed for 2-26 months (mean 14.6 &#45 6.3) post-treatment. The recurrence rates at 1 and 2 years were 20.4% and 28.1% within the liver in treatment group, respectively. The survival rates at 1 and 2 years were 92.5% and 75.3% for the treatment group. T-cells, NK cells and macrophages increased significantly in both treated and untreated nodules after PMCT, albeit less markedly within untreated nodules when compared to the treated ones. There is a statistically significant correlation between survival outcome and the extent of immunocyte infiltration. Conclusions: For inoperable HCC patients, PMCT is one of the treatment choices shown to be effective. Apart from its tissue coagulation effect, an increased systemic immune response directed against the tumour may also play an important role in improved survival.     

Surgical Significance of Telomerase Activity in Noncancerous Liver Tissue from Patients with Hepatocellular Carcinoma

Telomerase activity has been detected in tissue from noncancerous liver of patients with chronic liver disease, but its functional significance remains to be elucidated. We therefore evaluated the telomerase activity in surgically obtained noncancerous liver tissue from 20 hepatocellular carcinoma (HCC) patients. Two samples of noncancerous liver tissue were obtained from each patient: one from the parenchyma adjacent to the HCC nodules of the resected specimen; the other from the parenchyma distant from the HCC nodules of the remnant liver. Telomerase activity was assayed by a non-radioisotope quantitative system based on "TRAP-eze.'Five samples from the noncancerous liver tissue adjacent to the HCC nodules (25.0%) were telomerase-positive; all such cases showed high-grade malignant potential, such as intrahepatic metastasis and/or portal vascular invasion and infiltration of the fibrous capsule in the corresponding HCC nodules, and telomerase positivity showed neither a relationship with the histological activity index scores nor a correlation with liver function. Interestingly, no telomerase activity was detected in any of the 20 samples obtained from the parenchyma of the remnant liver. These results indicate that telomerase in noncancerous liver tissue is associated not with the hepatic condition accompanying HCC, but with the biological characteristics of the tumor itself, and may derive from infiltrating cancer cells. Determination of telomerase status may aid in designing more effective surgical procedures.