腹部肿瘤病人rIL-2/LAK治疗前后T细胞亚群、淋巴细胞转化试验及NK细胞活性变化

本文对30例晚期腹部肿瘤患者rIL－2／LAK治疗前后的免疫功能进行动态观察。检测指标包括T细胞亚群（CD3、CD4、CD8及CD4／CD8）。淋巴细胞转化试验（LymphocytesBlastogenesisTestLBT）和NK细胞活性。结果表明：肿瘤患者经rIL－2／LAK治疗后，CD3、CD4明显升高．CD8也略有升高．CD4／CD8明显升高；淋巴细胞转化率明显升高；经rIL－2／LAK治疗后，NK细胞活性增强。这反映肿瘤患者经rIL－2／LAK治疗后机体细胞免疫功能得到增强。     

肝癌患者过继免疫与栓塞化疗治疗前后免疫功能变化

对46例原发性肝癌患者LAK／IL－2肝动脉灌注与栓塞化符合用及单纯栓塞化行治疗前后机体免疫状态，即T淋巴细胞亚群，可溶性白细胞介素-2受体（sIL－2R），TNF及自身LAK细胞活性进行了比较研究。结果表明，治疗前患者T淋巴细胞亚群CD3、CD4明显低于正常对照组。CD3升高，CD4／CD8比值下降；sIL－2R及TNF水平明显高于正常对照组。治疗后，LAK／IL－2肝动脉灌注即过继免疫与栓塞化疗合用组，T淋巴细胞亚群CD3、CD4升高（P＜0.01），CD3下降（P＜0．01）、CD4／CD8比值升高（P＜0．01）sIL－2R水平下降（P＜0.01），TNF水平亦下降，自身LAK细胞活性增强（P＜0.01）；单纯栓塞化疗组上述指标无显著变化。     

肺癌和肝癌病人细胞免疫功能状态的研究

近年来,对有关原发性支气管肺癌(简称肺癌)和原发性肝癌(简称肝癌)的临床免疫学研究,发现均存在着细胞免疫低下,T细胞亚群比例失衡及兔疫监视功能缺陷。本文对肺癌和肝癌病人探讨NK、LAK与T细胞亚群之间的关系。 实验结果表明肺癌、肝癌患者NK及LAK细胞活性明显低于正常人,差异非常显著。经方差分析,肺癌各病理分型间NK及     

A-NK细胞的抗肿瘤作用

过继免疫治疗（AIT）是肿瘤生物治疗中的主要疗法。20世纪80年代起，LAK、TIL、CIL、CD3AK等效应细胞陆续被研究，国内外基础研究和临床应用报道层出不穷。1988年Vujanovic首次从LAK细胞中分离纯化了一种具有高效杀伤活性的LAK亚群，因其具有黏附在塑料表面的特征，故命名为A-LAK（adherent LAK）。20世纪90年代后以A—NK（IL-2 activated NK cell）代替了A-LAK。本实验参照文献简便快速地制备A-NK细胞，并在动物肝癌模型研究其抗肿瘤作用。     

生物反应调节剂在提高LAK细胞抗肿瘤活性应用

自1982年美国学者Rosenberg等发现淋巴因子激活的杀伤细胞（LAK细胞）以来,淋巴因子激活的杀伤细胞／白介素2（IL－2）过继免疫疗法对晚期肿瘤病人的治疗取得了确切的疗效,是肿瘤过继免疫治疗的重大突破,但由于IL－2的用量较大,常导致严重的毛细血管渗漏综合症（CLS）等副作用的发生,从而限制了LAK／IL－2疗法的广泛应用。近十年来,由于新的生物反应调节剂（BRM）的出现．给LAK／IL－2肿瘤过继免疫治疗带来新的生机。根据Mihich的定义：BRM是指能通过调整宿主对肿瘤的反应使二者之间的相互作用朝向有利于治疗肿瘤的方向发…     

抗肝癌iRNA及S-TF对肝癌患者LAK活性、IL-2活性及IL-2R表达的调节效应

 观察抗肝癌iRNA及S-TF对原发性肝癌（PHC）患者外周血LAK、IL－2活性及IL－2R表达的影响。发现PHC病人LAK、IL-2活性及IL-2R表达均明显低于正常人，且IL-2与LAK间呈正相关；经抗肝癌iRNA及S-TF治疗后，PHC病人的上述指标及WBC数均较治疗前显著升高。提示PHC病人的免疫功能低下或紊乱可能与肿瘤的发生及发展有关，应用抗肝癌iRNA及S-TF可给肿瘤宿主转移介导其细胞免疫功能，改善其免疫状态。     

原发性肝癌过继免疫治疗现状

原发性肝癌的过继免疫治疗是近年随着免疫效应细胞体外培养成为现实而发展起来的一种新的治疗手段.本文就原发性肝癌LAK细胞和TIL细胞过继免疫治疗的输入途径、体内分布、癌瘤大小与自体LAK活性的关系、治疗效果作一介绍与评述,并就其影响因素、存活的问题以及发展的方向提出了自己的浅见.     

肝动脉灌注LAK／IL—2加栓塞治疗原发性肝癌的疗效评价

近年来，应用LAK细胞及IL－2治疗人体恶性肿瘤的研究日益受到国内外学者的重视［1～3］。我们行肝动脉灌注LAK／IL－2加栓塞治疗原发性肝癌31例，并以同期化疗栓塞的29例作对照，取得了满意效果，总结报告如下。1材料与方法本文搜集近2年间无手术指征的原发性肝癌60例，并随机将其分成两组。行肝动脉灌注LAK／IL－2加栓塞者31例为A组，行化疗栓塞者29例为B组。肝动脉造影完成后，A组患者注入Lipiodol（LP）10～25ml、IL－230万单位及LAK细胞悬液100ml（约109个LAK细胞），其中7例追加适量明胶海绵颗粒栓塞。术后10天每日皮下补…     

多细胞过继性免疫治疗治疗晚期恶性肿瘤的近期疗效及安全性分析

目的:初步观察DC、CIK、γδT、CD3AK及NK细胞多细胞过继性免疫治疗对晚期恶性肿瘤患者的近期临床疗效及安全性分析。方法:对我院在2014年10月至2015年10月收治的162例次晚期恶性肿瘤患者进行多细胞过继性细胞免疫治疗,血细胞分离机单采自体外周血单个核细胞（peripheral blood mononuclear cells,PBMC),体外培养扩增DC、CIK、γδT、CD3AK及NK细胞,采用FCM法检测各细胞表型,并按计划序贯回输给患者进行治疗。结果:69例（43%）患者临床症状有不同程度改善,尤其是精神、食欲及睡眠质量改善最为明显,安全性分析显示主要不良反应为发热,占15%,28例完成3疗程免疫治疗患者瘤体变化为部分缓解（partial remission,PR）2例,疾病稳定（stable disease,SD）16例,疾病控制率（disease control rate,DCR）为64%。结论:DC、CIK、γδT、CD3AK及NK细胞过继性免疫治疗对晚期恶性肿瘤安全有效,可明显改善其生活质量,具有一定的临床应用前景。     

DC／CIK细胞在非小细胞肺癌治疗中的研究进展

树突状细胞／细胞因子诱导的杀伤细胞（dendritic cells／cytokine－induced killer cells,DC／CIK）是新型的异质性免疫效应细胞群,其主要的效应细胞同时表达CD3＋和CD56＋两种膜蛋白分子,兼有T淋巴细胞强大的抗肿瘤活性和自然杀伤（natural killer,NK）细胞的非主要组织相容性复合体（non－major histocom-patibility complex,MHC）限制性杀瘤特点。其与手术、放疗、化疗等方法相结合治疗非小细胞肺癌（non－small cell lung cancer,NSCLC）,可以使患者获得更高的生存率,显著提高患者的生活质量。DC／CIK由于其强大的体外扩增能力和抗肿瘤活性而迅速成为目前过继免疫治疗的有效方法之一。     

Long-term prognosis of hepatocellular carcinoma patients treated with adoptive immunotherapy

Adoptive immunotherapy was performed for patients with multiple hepatocellular carcinoma (HCC) using LAK cells via the hepatic arterial catheter. One case study: A 45-year-old male patient was performed an absolute-non-cure operation. The operation was a success. However, some residual tumors remained in the liver. After the operation, adoptive immunotherapy was performed for 7 times in 5 weeks. The total inoculated LAK cells were 6.9x10(9). After the adoptive immunotherapy, no residual tumors were detected in the liver. He survived for 7 years in remission. Our results show that adoptive immunotherapy with LAK cells is useful and effective for patients with multiple HCC.     

基于HLA-E功能特点的肿瘤免疫治疗新策略

人类白细胞抗原-E（HLA-E）在多种肿瘤中呈高表达,其与肿瘤患者预后的关系呈现肿瘤类型依赖性。HLA-E主要通过与NK细胞或T细胞上的激活性（NKG2C）或抑制性（NKG2A）受体结合,在调控抗肿瘤免疫应答中发挥重要作用。基于此,靶向 HLA-E/NKG2A 以阻断 HLA-E 与 NKG2A 的相互作用或抑制 HLA-E 表达,有望成为增强抗肿瘤免疫应答的新策略。鉴于HLA-E的功能特点设计增强型或通用型的T/NK细胞过继免疫疗法,有望提高过继免疫细胞疗法的治疗效果,具有良好的研发和临床应用前景。如何将靶向HLA-E或NKG2A的疗法与其他免疫疗法有效联合,实现更精准的免疫治疗以提高临床治疗效果是目前该方面研发和应用需要解决的难题之一。     

过继性免疫治疗对肺癌围术期细胞免疫功能的影响

对６９例可手术的非小细胞肺癌病例，随机分组分成ＬＡＫ细胞＋ＩＬ－２治疗的研究组２３例，单纯手术的对照组４６例，采用多元方差和单因素方差的统计方法，分析术后辅助性ＬＡＫ细胞过继性免疫治疗对围术期非小细胞肺癌细胞免疫功能的影响。结果显示，对ＣＤ４，ＣＤ４／ＣＤ８比率、ＮＫ细胞活性和淋巴细胞转化率４个指标的综合分析，两组间在围术期不同时间段上的细胞免疫功能都无显著性差异（各组间的ＨｏｔｅｌｉｎｇＴ２检验，Ｐ值均大于０．０５），但研究组术后１２ｄ的ＮＫ细胞活性较术前下降（单因素方差分析Ｐ＜０．０５），与对照组比较，差异也有统计学上的显著性意义（多元方差分析Ｆ＝４．５７１１，Ｐ＜０．０５）。     

Autologous tumor specific immunotherapy of refractory cancers with ex vivo-generated T cells stimulated by autologous tumor cell

OBJECTIVES: Autologous tumor-cell stimulated cytotoxic T lymphocytes (AuTLs) were prepared from peripheral blood T cells and the T cells were activated ex vivo over a 2-week culturing process in the presence of IL-2 and autologous biopsied tumor-cells from advanced cancer patients. These AuTLs may have potential for efficacy as a locoregional therapy in patients with refractory cancer. METHODS: Twenty-nine of 35 cancer patients (13 esophagus, 5 lung, 3 stomach, 4 breast, 1 melanoma) were enrolled in an early Phase II clinical trial. The patients received direct locoregional intratumoral injection of AuTLs biweekly through medical endoscope or intraarterial infusion reservoir system. Mean 0.25 x 10(9) AuTLs were injected 1 x /2 weeks for 6 weeks. RESULTS: Adverse events related to AuTL were minimal. AuTLs specific for autologous tumor cells were observed in 12 of 29 patients. Seven of these 12 patients (58.3%) had partial response (PR) or stable disease (SD). In contrast, 8 of 23 (34.8%) remaining patients treated by non-specific AuTLs had SD. Infiltration of T effector cells was significantly increased on the biopsied tumor specimens in the immunohistological studies. Furthermore, 11 patients receiving systemic infusion of non-specific LAK/NK T cells without autologous tumor-cell stimulation only had 2 SDs (18.2%). CONCLUSIONS: Our results demonstrated the clinical potential of intra-peritumoral administration of AuTLs. Thus, locoregional immunotherapy with autologous tumor specific AuTLs may be more effective than systemic adoptive immunotherapy using intravenous infusion of autologous tumor non-specific LAK/NK T cells for the treatment of solid cancers.     

LPAK细胞的增殖效应及其初步临床应用

用ＬＰＡＫ（ＬｙｍｐｈｏｋｉｎｅａｎｄＰＨＡＡｃｔｉｖｉａｔｅｄＫｉｌｌｅｒ）细胞／ＩＬ－２过继免疫疗法治疗１７例晚期肿瘤，在１２例可评估病例中，５例肝癌（包括转性肝癌１例）有３例肿瘤分别缩小３５．７％、３９．５％和４７．８％；２例肺癌和２例转移性肺癌治疗后肿瘤无明显变化；１例胃癌癌性腹水治疗后腹水大部消失；１例皮肤恶性淋巴瘤治疗后皮损消退５０％以上。     

Cancer treatment by photodynamic therapy combined with adoptive immunotherapy using genetically altered natural killer cell line.

Adoptive immunotherapy mediated by human natural killer (NK) cell line genetically altered to produce interleukin-2 (NK92MI) was evaluated as adjuvant to photodynamic therapy (PDT) of subcutaneous tumors. The combined effect of these two modalities was first examined with SiHa tumors (human cervical squamous cell carcinoma) growing in NOD-scid mice. The most effective protocol for NK92MI cell transfer in conjunction with PDT mediated by photosensitizer mTHPC was the injection of 5 x 10(7) cells (peritumoral or intravenous) given immediately after PDT, which produced a marked improvement in the therapeutic outcome compared with the effect of PDT alone. The same protocol was tested with HT-29 tumor model (human colorectal adenocarcinoma) xenografted in NOD-scid mice. The results demonstrate that the adoptive immunotherapy with NK92MI cells (which when used alone were not effective in controlling tumor growth) significantly improved the cures of PDT-treated HT-29 tumors, whereas such benefit was not observed with the parental cell line NK92 (not producing interleukin-2). Flow cytometry-based analysis revealed a higher percentage of p.t. injected NK92MI cells in PDT-treated than in non-treated HT-29 tumors. Further investigation showed that the NK92MI cell-based adoptive immunotherapy is also a highly effective adjuvant for PDT treatment of murine EMT6 tumors growing in immunocompetent syngeneic BALB/c mice. This result diminishes the concern that adoptively transferred NK92MI cells may be rendered ineffective by an allogenic reaction of the host. The findings of this study suggest that advanced protocols of NK cell-based adoptive immunotherapy can be developed as efficient adjuvants to PDT used for the treatment of solid malignant tumors.     

Adoptive immunotherapy using lymphokine-activated killer (LAK) cells and interleukin-2 for recurrent malignant primary brain tumors

Summary Ten patients with recurrent malignant primary brain neoplasms were treated with adoptive immunotherapy using lymphokine-activated killer (LAK) cells and interleukin-2 (IL-2). Nine patients had supratentorial glioma and they received multiple intratumoral instillations of LAK cells through reservoir-catheter system or burrhole. The other patients with disseminated subarachnoid metastases from posterior fossa medulloblastoma received immunotherapy via lumbar subarachnoid route. A partial and transient clinical response was observed in two patients following the therapy, and a cystic transformation of the essentially solid tumor was noted on the CT scans of these two patients. No significant clinical or radiological response to the treatment was observed in the remaining 8 patients. The results of this preliminary study reveal limitations of the regional intratumoral adoptive immunotherapy using currently available techniques and provide sufficient evidence of its effectiveness to warrant further investigations.     

肝细胞肝癌对机体细胞免疫的影响

目的　探讨原发性肝细胞肝癌(HCC )对机体细胞免疫机能的影响。方法　采用血常规检测,应用荧光标记抗体及三色激光流式细胞仪对5 1例肝细胞肝癌患者和17例对照样本术前、术后外周血的细胞免疫指标进行检测。检测项目包括:T辅助淋巴细胞(THL) ,细胞毒T淋巴细胞(CTL) ,NK细胞,单核细胞,并进行手术前后的对比分析。结果　原发性肝细胞肝癌(HCC)术后THL、CTL、NK和单核细胞百分比、单核细胞绝对值的平均值较术前分别增高0 .91% ,8.5 8% ,8.85 % ,3 1.45 % ,66.5 5 % ,手术前后有显著性差异(P <0 .0 5或P <0 .0 1)。良性对照组的术前与术后上述各项指标比较,均无显著性差异(P >0 .0 5 )。结论　肝细胞肝癌(HCC)对机体的细胞免疫产生明显抑制作用,在肿瘤切除之后,这种抑制能力随之减弱,细胞免疫得到恢复。而良性对照组对机体细胞免疫的抑制不显著。综合分析细胞免疫的变化情况,对判断HCC的预后及肿瘤复发有重要的意义。     

Immune responses of dendritic cells combined with tumor-derived autophagosome vaccine on hepatocellular carcinoma

Background

To induce and collect tumor-derived autophagosomes (DRibbles) from tumor cells as an antitumor vaccine by inhibiting the functions of proteasomes and lysosomes.

Methods

Dendritic cells (DCs) generated from peripheral blood mononuclear cell (PBMC) of hepatocellular carcinoma (HCC) patients were cocultured with DRibbles, and then surface molecules of DCs, as well as surface molecules on DCs, were determined by flow cytometry. Meanwhile, immune responses of the DCs-DRibbles were examined by mixed lymphocyte reactions.

Results

DRibbles significantly induced the expression of CD80, CD83, CD86 and HLA-DR on DCs. The enzyme-linked immunosorbnent assay (ELISA) showed that IFN-γ levels after vaccination increased than before in most patients, but CD8+ proportion of PBMC increased only in nine patients. Higher levels of IFN-γ were detected in the CD8+ cells than CD4+ T cells. These results suggested that DCs-DRibbles vaccine could induce antigen-specific cellular immune response on HCC and could prime strong CD8+ T cell responses, supporting it as a tumor vaccine candidate.

Conclusions

Our results demonstrate that HCC/DRibbles-pulsed DCs immunotherapy might be deployed as an effective antitumor vaccine for HCC immunotherapy in clinical trials.