肿瘤浸润淋巴细胞及重组白细胞介素2治疗原发性肝细胞癌

目的探讨联合应用肿瘤浸润性淋巴细胞（ｔｕｍｏｒｉｎｆｉｔｒａｔｉｎｇｌｙｍｐｈｏｃｙｔｅｓ,ＴＩＬ）和重组白细胞介素２（ｒＩＬ２）治疗对原发性肝细胞癌（ｐｒｉｍａｒｙｈｅｐａｔｏｃｅｌｕｌａｒｃａｒｃｉｎｏｍａ,ＰＨＣＣ）术后患者细胞免疫功能的影响和其临床疗效。方法对１６例ＰＨＣＣ术后患者进行了ＴＩＬ和ｒＩＬ２治疗,１２例从肝动脉插管输入,４例从门静脉插管输入。结果ＴＩＬ治疗后１６例患者外周血白细胞介素２（ＩＬ２）,Ｔ细胞亚群及比率均有不同程度上升;肿瘤坏死因子（ＴＮＦ）,白细胞介素６（ＩＬ６）多有不同程度下降。随访６～２４个月,１４例肝癌根治性切除术患者,肿瘤无复发迹象,１例肝癌根治切除术患者ＴＮＦ上升了８ＩＵ,该患者治疗８个月后死于肝功能衰竭;１例肝癌姑息性切除,残癌无水酒精注射病人,ＴＩＬ治疗１８个月后肿瘤直径扩大２ｃｍ。结论ＴＩＬ治疗对提高ＰＨＣＣ术后患者抗肿瘤细胞免疫功能,防止肿瘤复发,抑制残瘤生长,延长患者生存期具有一定作用。     

肿瘤浸润淋巴细胞、重组白细胞介素2治疗对手术后原发性肝癌病人细胞免疫功能的影响

对５例经手术切除的原发性肝癌病人用肿瘤浸润淋巴细胞（ＴＩＬ）和重组白细胞介素２（ｒＩＬ－２）治疗。治疗前１周及治疗后１个月检测细胞免疫功能，治疗后外周血淋巴细胞的ＮＫ活性、白细胞介素２产生及活性均明显升高；植物血凝素（ＰＨＡ）皮试阴性转阳性。提示；ｒＩＬ－２和ＴＩＬ应用可明显提高原发性肝癌病人的细胞免疫功能，对杀灭残留癌细胞，抗肿瘤复发、转移，提高远期疗效有一定作用。可能成为肝癌术后抗肿瘤复发，转移的重要疗法。     

可溶性白细胞介素2受体（sIL—2R）与大肠癌

恶性肿瘤血清可溶性白细胞介素２受体（ｓＩＬ－２Ｒ）水平增高是肿瘤诱导的免疫反应所致。大肠癌血清ｓＩＬ－２Ｒ水平明显比正常人群高，且与大肠癌临床分期有关，根治性大肠癌切除术后２周ｓＩＬ－２Ｒ可降至正常水平，研究表明ｓＩＬ－２Ｒ水平与大肠癌患者的免疫状态及病情严重程度有关。     

重组人白细胞介素－2对恶性肿瘤疗效的观察

对２５例恶性肿瘤患者单独或配合化疗使用重组人白细胞介素－２（ｒＩＬ－２）的疗效进行临床观察。２５例中男性１５例，女性１０例。平均年龄５３．１岁。肺癌７例，乳腺癌５例，结肠癌４例，直肠癌３例，肝癌、胃癌各２例，肾癌、神经纤维肉瘤各１例，分别采用全身静脉、瘤内注射、体腔内注射、介入动脉灌注等途径给药。观察结果：完全缓解２例，部分缓解６例，稳定２０例，总有效率为２８．６％。表明ｒＩＬ－２对改善恶性肿瘤患者的生活质量有一定帮助与化疗配合能提高疗效，且毒副反应轻。     

白细胞介素(IL)-2与IL-4对小鼠膀胱癌肿瘤浸润性淋巴细胞增殖及细胞毒性的协同调节作用

目的 探讨白细胞介素（ＩＬ）－２与ＩＬ－４对膀胱癌肿瘤浸润性淋巴细胞（ＴＩＬ）体外增殖及细胞毒性免疫调控的协同作用。方法 分离膀胱癌ＴＩＬ,置于含ＩＬ－２和（或）ＩＬ－２的完全培养基因中培养４周,定期计数ＴＩＬ增殖数量。四甲基偶氮唑蓝（ＭＴＴ）比色法检测ＴＩＬ细胞毒性。结果 对比单纯ＩＬ－２的培养条件,ＩＬ－２联合ＩＬ－４后４周时ＴＩＬ扩增数量是前者的１．６５倍（Ｐ〈０．０５）。在交靶比为１０：１时,ＴＩＬ对自体膀胱癌细胞（ＢＴＴ７３９）表现出高水平的杀伤活性（Ｐ〈０．０５）。联合培养的ＴＩＬ抗ＢＴＴ３９或小鼠淋巴瘤瘤株（ＹＡＣ－１）的活性与在单纯ＩＬ－２培养的条件下相比无显著改变（Ｐ均〉０．０５）。结论 ＩＬ－４对ＩＬ－２活化的膀胱癌ＴＩＬ增殖具有较强的正向调节效应,而对ＴＩＬ细胞毒性未见明显影响。     

可溶性白细胞介素2受体(sIL-2R)与大肠癌(文献综述)

恶性肿瘤血清可溶性白细胞介素2受体（SIL－2R）水平增高是由肿瘤诱导的免疫反应所致。大肠癌血清SIL－2R水平明显比正常人群高，且与大肠癌临床分期有关，根治性大肠癌切除术后2周SIL－2R可降至正常水平。研究表明sIL－2R水平与大肠癌患者的免疫状态及病情严重程度有关。     

膀胱癌TIL的分离培养及其表型分析

对３例膀胱移行细胞病患者手术切除的肿瘤组织中肿瘤浸浸润性淋巴细胞进行了体外分离与培养，结果２例获得成功，ＴＩＬ体外扩增培数分别达５８－１７０，在培养２４ｄ时对自体肿瘤靶细胞的最高杀伤活性达４１％以上，且呈现一定的靶细胞特异性。     

原发性肝癌患者肿瘤浸润T淋巴细胞亚群分析

目的:了解原发性肝癌患者肿瘤浸润T淋巴细胞中CD4+及CD8+的表达特点。方法:收集30例原发性肝癌患者(均有乙型肝炎病毒感染背景)的癌组织(乙肝肝癌组)和癌旁组织(乙肝癌旁组)以及30例因良性病变而行肝切除的患者的新鲜肝组织(对照组),分离组织浸润淋巴细胞,用抗CD3、CD4和CD8单克隆抗体同时荧光染色,用流式细胞仪检测各表面标志的表达情况。结果:1)乙肝肝癌组、乙肝癌旁组及对照组CD3+CD4+T细胞占浸润淋巴细胞的比例分别为(22.31±3.68)%、(10.69±2.47)%及(4.21±4.26)%。乙肝肝癌组显著高于乙肝癌旁组及对照组,差异有统计学意义(P=0.000,P=0.001),乙肝癌旁组高于对照组,差异亦有统计学意义(P=0.019)。2)乙肝肝癌组、乙肝癌旁组及对照组CD3+CD8+T细胞占浸润淋巴细胞的比例分别为(26.10±5.82)%、(21.82±2.70)%及(41.31±14.01)%,乙肝肝癌组及乙肝癌旁组显著低于对照组,差异有统计学意义(P=0.014,P=0.004),而乙肝肝癌组与乙肝癌旁组之间差异无统计学意义(P=0.651)。3)乙肝肝癌组组织浸润淋巴细胞中CD3+CD4+T细胞与CD3+CD8+T细胞的比值为0.91±0.30,显著高于乙肝癌旁组(0.47±0.11,P=0.003)及对照组(0.11±0.13,P=0.000),CD3+CD4+与CD3+CD8+T细胞的比值出现失衡。结论:原发性肝癌患者肿瘤浸润淋巴细胞中T细胞亚群失调,表现为CD3+CD4+T细胞所占比例升高,CD3+CD8+T细胞所占的比例下降,CD3+CD4+/CD3+CD8+明显升高。     

肿瘤浸润淋巴细胞在肾细胞癌原位表达的特点

目的:研究肾细胞癌组织中肿瘤浸润淋巴细胞(TIL)的分布特点及其与预后的关系.方法:对52例肾透明细胞癌组织T,辅助T及自然杀伤细胞(NK细胞)进行原位免疫组化研究,并与病人预后作相关性分析.结果:TIL细胞中主要为T细胞,占70%;TIL细胞表达的程度对肾细胞癌患者预后的影响较大(P<0.01);TIL细胞亚群之间的表达为正相关.结论:TIL在对抗肾细胞癌中起较重要的作用,它们的表达程度影响预后.     

Chemosensitivity,tumor infiltrating lymphocytes (TILs), and survival of postpartum PABC patients treated by neoadjuvant chemotherapy

BackgroundPregnancy-associated breast cancer (PABC) refers to breast cancers (BC) diagnosed during pregnancy or shortly after birth. Although the inflammatory environment of post-partum PABC cases (designed as PP-PABC) may be deleterious, so far PP-PABC have scarcely been distinguished from breast cancers diagnosed during pregnancy. Furthermore, whether PP-PABC cases have an enhanced immune infiltration remains unknown. We investigated chemosensitivity, immune infiltration and survival of PP-PABC patients treated by neoadjuvant chemotherapy (NAC) compared to non-PABC matched BC patients.Materials and methodsWe identified PP-PABC cases among a cohort of 1199 invasive BC treated with NAC between 2002 and 2012. Each PP-PABC case was matched with 3 non-PABC controls, according to age and pathological breast cancer subtype. Microbiopsy specimens and paired surgical samples were evaluated for stromal lymphocyte infiltration. Association of clinical and pathological factors with pathological complete response (pCR) and disease-free survival (DFS) was assessed by univariate and multivariate analyses.ResultsOur final population study was composed of 116 patients (29 PP-PABC and 87 non-PABC). Median follow-up was of 49.0 and 29.3 months, respectively. After NAC, pCR rates (p = 0.64), post-NAC immune infiltration (stromal TILs: p = 0.67; intratumoral TILs: p = 0.14), and DFS rates (p = 0.17) were comparable between PP-PABC and non-PABC patients in global population. Similar results were found after stratification by pathological subtype.ConclusionWe observed similar patterns between postpartum PABC and control tumors in terms of chemosensitivity, immune infiltration, and prognostic. Our results enhance the idea that PP-PABC should receive the same standard of care treatment as other patients, including neoadjuvant chemotherapy.     

Unfavorable prognostic role of tumor-infiltrating lymphocytes in hormone-receptor positive,HER2 negative metastatic breast cancer treated with metronomic chemotherapy

BackgroundHigh levels of tumor-infiltrating lymphocytes (TILs) in primary triple negative and HER2-positive breast cancer (BC) have been associated with an improved patients' outcome. The role of TILs in Luminal (hormone receptor positive and HER2 negative) tumors remains to be elucidated. Moreover, the association between TILs and prognosis in the metastatic setting is still unknown.Patients and methodsWe evaluated the relationship between TILs and time to progression (TTP) in metastatic BC patients enrolled in a prospective phase II trial of metronomic chemotherapy, that used cyclophosphamide 50 mg daily, capecitabine 500 mg thrice daily and vinorelbine 40 mg orally three times a week (VEX combination).ResultsOf the 108 ER + BC patients enrolled in the VEX trial, 92 (85%) had sufficient tumor tissue and were assessed for TILs in H&E stained slides. TILs were evaluated in 38 primary BC samples and 54 metastatic sites. High (≥10%) TILs levels were significantly correlated with high Ki-67 labeling index. At multivariable analysis, each 10% increase in TILs strongly predicted a worse TTP (HR: 1.27, p = 0.008). VEX trial patients, categorized by a 3 tiers system (0–4%, 5–9% and >10% TILs) showed significantly different progression free survival curves (p = 0.011).ConclusionsHigh TILs levels are significantly associated with a worse TTP in Luminal metastatic BC patients treated by metronomic chemotherapy. Our data confirm the reliability of TILs as a biomarker in the BC metastatic setting. The putative unfavorable prognostic role of TILs in Luminal BC patients might have clinical utility if validated by further studies.     

Regulatory effects of interleukin-7 on renal tumor infiltrating lymphocytes

Summary Biological therapy using a combination of lymphokine and tumor infiltrating lymphocytes (TILs) is a new approach to the treatment of patients with advanced cancer. To improve the potency of TILs, new cytokines with T-cell stimulatory effects used alone or in combination with interleukin-2 (IL-2) are currently being investigated. We have studied the effect of interleukin-7 (IL-7) on TILs derived from renal cell carcinoma. Our data demonstrated that five of ten TILs proliferated in response to IL-7 alone. This proliferative response was 73–90% less than that obtained with IL-2 alone. The use of IL-7 plus IL-2 resulted in a 1.2- to 4.7-fold increase in proliferation of six of ten TILs compared with IL-2 alone. IL-7-driven TIL growth was consistently blocked by anti-IL-2, anti-IL-2R and anti-IL-7 antibodies (37.2%, 41.6% and 82.2% suppression, respectively). The expression of IL-2 receptors was also significantly increased in the presence of IL-7 or IL-7 phytohemagglutinin (40.6+3.8 and 72.5+1.5). In comparison with IL-2, IL-7 treatment was associated with a decrease in CD56 (46.3%±19 vs 10%±4.9) and increase in CD3 (29.3%±12 vs 73%±6.4) and CD 4 (19.3%±15 vs 58%±10). These studies suggest that in some renal TILs, IL-7 and IL-2 can have a synergistic proliferative effect. The IL-7 stimulatory effect appears to be mediated via both an IL-2 pathway and an IL-7-independent pathway.This paper was selected for publication in Urological Research from the program of the 1991 meeting of the European Society of Urological Oncology and Endocrinology (ESUOE)     

肿瘤浸润性淋巴细胞治疗原发性肝癌的基础与临床研究

目的 探讨肿瘤浸润性淋巴细胞（TIL）对原发性肝癌（HCC）患者术后细胞免疫功能的影响和临床疗效及半乳糖抗CD3单克隆体－TIL复合物（Gal-Anti-CD3-McAb-TIL)体外及体内的趋肝性。方法 （1）对30例HCC患者术后进行了TIL和重组白介素－2（rIL-2)治疗。（2）制备半乳糖基抗CD3单克隆抗体－TIL（McAb-TIL)复合物。将TIL和McAb－TIL分别与小鼠肝细胞置CO2孵箱中孵育,用倒置相差显微镜观察其趋肝性。将^125I－TIL和^125I-McAb-TIL分别经小鼠尾静脉注入,观察示踪剂在脏器中的分布。结果 TIL治疗后,30例患者外周rIL-2、T细胞亚群均有上升。24例肝癌根治性切除术患者随访60－48个月无复发。McAb－TIL与细胞有明显粘附,给小鼠静脉注射后,TIL和McAb-TIL主要在肺中积聚,在肝内浓度较低。结论 TIL治疗可能提高HCC患者术后抗钟瘤细胞免疫功能。TIL与半乳糖抗CD3单克隆抗体结合物体内肝靶向性不明显。     

Effect of pre-operative short course famotidine on tumor infiltrating lymphocytes in colorectal cancer: a double blind, placebo controlled, prospective randomized study

BACKGROUND: Pilot studies have shown that histamine H2 receptor antagonists augment the natural immunity against cancer in colorectal and gastric cancer by enhancing lymphocytic infiltration in the tumors. However, a study of adjuvant ranitidine failed to show a significant benefit in colorectal cancer, possibly because of the immunosuppression exerted by blood transfusion and post-operative infections. The pre-operative use of H2 receptor antagonists may therefore be of greater benefit. Except for a pilot study using cimetidine, there are no trials that have evaluated the effect of pre-operative H2 receptor antagonists on tumor infiltrating lymphocytes in colorectal cancer. OBJECTIVE: To evaluate the efficacy of famotidine in augmenting tumor infiltrating lymphocytes in colorectal cancer. STUDY DESIGN: Double blind, placebo controlled, prospective randomized study. METHODS: Twenty-three patients with resectable colorectal cancer were randomized to receive famotidine (n = 11) or placebo (n = 12). Famotidine was given for 1 week pre-operatively in a dose of 40 mg per day p.o. After resection, the specimens were analyzed histologically for lymphocytic infiltration by a pathologist blinded to the two groups. Lymphocytic infiltration more than 50 cells per high power field, involving more than 50% of the tumor-normal tissue interface was considered significant. RESULTS: The two groups were comparable for age, gender, pre-operative carcino embryonic antigen (CEA) levels and pathological stage. Significant lymphocytic infiltration was seen in 63.6% (7 of 11) patients in the study group compared to only 8.5% (1 of 12) patients in the placebo group (P = 0.005). Despite fewer recurrences and a longer survival in the study group, the difference was not significant. CONCLUSION: This study shows that pre-operative famotidine may significantly enhance lymphocytic infiltration in colorectal cancer and may have potential for use as an anticancer agent in colorectal cancer.