超声引导下对晚期直肠癌患者行NKG2D-CAR基因治疗1例

正患者男,51岁,经标准治疗失败的转移性直肠癌患者,因入组"修饰性免疫细胞(NRC-NK)个体化治疗Ⅳ期结直肠癌的临床研究"入院。影像学检查提示:患者肝周腹膜下多发转移灶伴左腹局限性无回声区,拟对患者行NRC-NK腹腔内回输,评估CAR-NK细胞的抗肿瘤治疗效果。第一次CAR-NK细胞回输过程中采用直接腹腔内注射给药,治疗效果欠佳。因此,第二次CAR-NK细胞回输前,超声介入组对患者情况进行评估后     

川芎嗪在自体血回输对NK细胞功能的影响

目的探讨川芎嗪对自体血回输患者NK细胞功能的影响。方法40例择期行脊柱手术患者,ASA分级Ⅰ～Ⅱ级,随机分成两组,即实验组和对照组,每组20例。实验组于收集血液前30min静脉滴注川芎嗪4mg／kg,在回收血液的肝素盐水和洗涤盐水内加入川芎嗪,终浓度为0．05mg／ml;对照组不予静滴川芎嗪,肝素盐水和洗涤盐水内不加川芎嗪。采集术前、回输自体血后1h、1d、5d的静脉血,使用流式细胞仪测定NK细胞（CD16＋56）的水平。结果对照组CD16＋56回输后1h明显升高,术后1d下降至术前水平,术后第5天比术前明显下降,实验组在回输后1h、1d较术前明显升高,至术后第5天仍在术前水平,在回输后第5天对照组明显低于实验组。结论对照组较实验组在回输自体血后NK细胞功能受到明显的抑制,表明川芎嗪在自体血回输中的应用对NK细胞功能具有一定的保护作用。     

NK细胞免疫治疗恶性肿瘤的研究进展

细胞免疫治疗是针对自身免疫细胞能力低下的恶性肿瘤患者进行的一种新型补充疗法,包括基于T细胞的嵌合抗原受体T细胞（CAR-T）疗法、肿瘤浸润淋巴细胞（TILs）疗法,基于NK细胞的嵌合抗原受体NK细胞（CAR-NK）疗法和自体细胞免疫疗法（CIK）,还有基于其他免疫细胞如单核吞噬细胞,包括树突状细胞和巨噬细胞的输注治疗。其中,自然杀伤细胞（nature killer cell,NK细胞）作为机体天然免疫的重要细胞,在机体抗肿瘤、抗病毒感染、免疫调节中发挥着重要作用。它可以像T细胞一样通过工程化改造后靶向治疗肿瘤,还能够进行同种异体来源的NK细胞输注治疗,弥补了T细胞自体来源受限和异体免疫排斥的缺点。研究证实,输注异体NK细胞的患者不会发生严重的移植物抗宿主病（graft versus hostdisease,GVHD）。N K细胞不仅扩大了用于细胞免疫治疗的细胞种类,还为形成较低成本的细胞免疫治疗产品提供了广阔应用前景。但是目前仍存在NK细胞质量不稳定,制备流程缺乏统一质量标准等问题,虽有部分NK细胞免疫治疗产品已经获得了美国食品药品监督管理局（Food and Drug Adminis...     

嵌合抗原受体修饰的CAR-T/NK细胞在多发性骨髓瘤治疗中的应用

嵌合抗原受体(chimeric antigen receptor,CAR)是一种合成跨膜蛋白,通过抗肿瘤细胞上的特异性或相关性抗原来重新定向被修饰的细胞。CAR修饰的T/NK细胞,特别是CAR-T细胞是近年来迅速发展的肿瘤过继免疫治疗新手段,它赋予T/NK细胞靶向杀伤活性,并可克服肿瘤局部免疫抑制微环境和打破宿主免疫耐受状态。CAR将识别肿瘤相关抗原的单链抗体和T/NK细胞的活化基序相结合,通过基因转导赋予T/NK细胞肿瘤靶向性、更强的杀伤活性和持久的生命力。本文就CAR发展、CAR-T和CAR-NK细胞的比较、MM细胞的表面标记和CAR在MM中的应用及CAR-T/NK细胞的前景进行综述。     

树突状细胞疫苗联合细胞因子诱导的杀伤细胞治疗鼻腔及鼻窦恶性黑色素瘤的近期疗效观察

目的探讨负载自体肿瘤抗原的树突状细胞(DC)疫苗联合细胞因子诱导的杀伤细胞(C IK)治疗鼻腔、鼻窦恶性黑色素瘤的疗效。方法对15例鼻腔鼻窦恶性黑色素瘤患者在常规手术后2周,通过体外培养扩增DC及C IK细胞,并静脉回输,进行免疫治疗,观察患者临床症状改善及生活质量改善状况及近期疗效,利用流式细胞仪检测回输前后患者T细胞亚群及NK细胞变化。结果15例接受自体DC及C IK细胞治疗患者中,治疗前后患者临床症状有明显改善,提高率达86%,自体回输免疫治疗后,CD3+、CD4+T细胞和NK细胞(CD16+、CD56+)及CD4+/CD8+比例均显著提高(P0.01)。结论负载自体肿瘤抗原的DC疫苗联合C IK治疗鼻腔、鼻窦恶性黑色素瘤可以提高患者免疫功能,改善临床症状,提高生存质量。     

回输自体来源的树突状细胞治疗慢性粒细胞白血病的临床研究——附3例报告

目的:观察体外诱导生成的自体慢性粒细胞白血病(慢粒)来源树突状细胞(dendriticcell,DC)体内回输的疗效、安全性及对体内免疫功能的影响.方法:3例慢粒慢性期患者(Ph染色体、BCR-ABL融合基因均为阳性)接受体外诱导的自体慢粒来源DC的静脉回输,DC回输数量为(4.1～5.9)×106,每周1次,共回输4次.通过检测回输前后迟发型超敏反应,回输前后外周血CD3 、CD4 、CD8 T细胞百分率的变化,以了解机体产生的免疫反应.回输前后定期检查患者的白细胞计数、骨髓细胞形态学、外周血Ph染色体阳性的细胞数、BCR-ABL融合基因阳性细胞所占比率,以观察慢粒的缓解情况.结果:3例患者在DC回输前迟发型超敏反应均为阴性,在第2次DC回输后呈阳性.DC回输后CD3 T细胞百分率明显升高,其中2例患者的CD4 T细胞百分率明显升高;在DC回输的过程中及之后的3个月,白细胞基本在正常值范围内,骨髓象为慢粒缓解期.Ph染色体阳性的细胞数有下降趋势,BCR-ABL融合基因阳性细胞所占比率有所下降.结论:回输自体来源DC激活了机体抗肿瘤免疫反应,可能具有促进慢粒缓解的作用,为慢粒的治疗提供了新的方法.     

DC-CIK自体回输对化疗后胃癌患者T淋巴细胞亚群及NK细胞的影响

目的探讨树突细胞-细胞因子诱导杀伤细胞(DC-CIK)自体回输对化疗后胃癌患者T淋巴细胞亚群及NK细胞的影响。方法前瞻性选取2016年2月至2017年2月陕西中医药大学附属医院胸外科收治的156例胃癌患者,采用随机数字表法将患者分为观察组和对照组,每组各78例。对照组予以单纯化疗治疗,观察组在此基础上辅以DC-CIK自体回输治疗。对比两组患者临床近期疗效,并分别于治疗前后对比两组患者外周血T淋巴细胞亚群及NK细胞的变化及治疗安全性。结果观察组患者有效率(RR)、疾病控制率(DCR)分别为47. 4%、66. 7%,对照组RR、DCR分别为41. 0%、51. 3%,两组患者RR比较差异无统计学意义(P 0. 05),两组患者DCR比较差异具有统计学意义(P 0. 05)。观察组患者治疗后外周血CD3~+(61. 27±3. 18%)、CD4~+(36. 45±4. 25%)、CD4~+/CD8~+(1. 55±0. 12%)、自然杀伤(NK)细胞百分比(21. 43±3. 17%)均高于对照组(53. 49±3. 75%、28. 17±3. 38%、1. 17±0. 15%、15. 28±3. 32%)(P 0. 05)。两组患者治疗期间均未发生严重不良反应。结论对晚期胃癌患者在化疗后辅以DCCIK自体回输治疗有助于提高T淋巴细胞亚群及NK细胞水平并有利于保障近期抗肿瘤的疗效。     

树突状细胞与细胞因子诱导的杀伤细胞免疫治疗对恶性肿瘤化疗后患者免疫功能的影响

目的探讨树突状细胞(DC)与细胞因子诱导的杀伤细胞(CIK)治疗对化疗后肿瘤患者免疫功能的影响。方法采集化疗后恶性肿瘤患者外周血单个核细胞,经体外诱导产生DC和CIK细胞,每个患者接受DC与CIK细胞治疗至少2个疗程;采用流式细胞术检测21例健康者和48例肿瘤患者化疗前后及DC联合CIK治疗后外周血淋巴细胞CD3+、CD4+、CD8+、NK细胞及CD4+/CD8+比值。结果肿瘤患者外周血CD3+、CD4+、CD8+、CD4+/CD8+及NK细胞数量低于健康对照组,差异具有统计学意义(P<0.05);化疗后外周血CD3+、CD4+、NK细胞及CD4+/CD8+比值较化疗前降低,差异均有统计学意义(P<0.05),但化疗前后CD8+均值变化无统计学意义(P>0.05)。DC与CIK细胞回输后外周血CD3+、CD4+、CD4+/CD8+及NK细胞均高于回输前,CD8+低于回输前,但CD3+、CD4+仍低于健康对照组,差异具有统计学意义(P<0.05)。结论恶性肿瘤患者化疗后免疫力降低,应用DC与CIK细胞免疫治疗,可提高恶性肿瘤患者的免疫功能。     

NK细胞的免疫生物学特性

人类NK细胞是机体抗肿瘤和抗病毒感染的天然免疫细胞，有自发的细胞毒功能，近10年来人们发现NK细胞的识别也具有特异性。NK细胞表达多种受体，按受体识别的分子可分为HLAI类分子特异性受体、非HLAI类分子特异性受体和共受体三大类，功能上划分为活化性受体和抑制性受体两类，这些受体对NK细胞的功能有重要的调节作用。本文就NK细胞受体的特点和功能作一综述，并探讨开发NK细胞新型潜在细胞免疫治疗的价值。     

基于NK细胞的血液系统肿瘤免疫治疗

自然杀伤(NK)细胞是先天性免疫系统的重要组成部分.它既是重要的外向性防御细胞,又是体内多种免疫细胞的调节细胞.探讨NK细胞在肿瘤发生发展过程中的作用及其作为免疫治疗的手段已受到人们的重视.笔者详细阐述了NK细胞的历史背景及抗肿瘤机制,对基于NK细胞的血液系统肿瘤免疫治疗进行了综述.     

β-榄香烯抗肿瘤免疫效应的研究进展

肿瘤免疫治疗能够激发或调动机体的免疫系统,增强肿瘤细胞免疫原性,从而达到控制和杀伤肿瘤细胞的目的。目前,肿瘤免疫治疗已经成为抗肿瘤研究的重要课题之一。中医药抗肿瘤免疫机制是多方面的,β-榄香烯是抗肿瘤代表中药莪术的有效提取物,本文综述了近年来β-榄香烯抗肿瘤免疫效应的临床及实验研究的进展情况。     

护理人员对抗肿瘤药物职业防护认知现状调查

[目的]了解我院临床一线护理人员对抗肿瘤药物职业防护的认知状况.[方法]采用自行设计的调查问卷对332名临床一线护理人员进行问卷调查.[结果]40%的人员参加过抗肿瘤药物使用和防护方法培训;不同职称护理人员对抗肿瘤药物防护知识的学习兴趣差异有统计学意义(P＜0.05);不同学历护理人员对抗肿瘤药物自身防护的了解程度差异有统计学意义(P＜0.05);不同接触频次护理人员对抗肿瘤药物自身防护知识的了解程度和学习兴趣差异有统计学意义(P＜0.01),但配药时戴手套情况差异无统计学意义(P＞0.05).[结论]要加强护理人员的职业防护教育,教会护理人员抗肿瘤药物的正确使用方法,建立抗肿瘤药物防护制度,并定期督促检查落实.     

Cooperative role of T lymphocytes and macrophages in anti-tumor activity of mice pretreated with schizophyllan (SPG)

Anti-tumor activity was studied in mice injected with Schizophyllan (SPG), a glucan produced by Schizophyllum commune Fries. SPG-injected mice rejected subcutaneously inoculated sarcoma-180 and the anti-tumor activity was shown to be mediated by host spleen and lymph node cells. The anti-tumor activity lasted as long as 60 days after the SPG administration as assessed by transfer of cells into normal recipients. Cells involved in anti-tumor activity were shown to be T lymphocytes since anti-tumor activity was diminished when lymph node cells from SPG-treated mice were treated with anti-Thy-1.2 sera plus complement, whereas cells passed through a nylon wool column retained the activity. Macrophages were also shown to be involved since administration of carrageenan or trypan blue into the host decreased the inhibition ratio of tumor growth. It was concluded that anti-tumor activity in SPG-treated mice was mediated by the cooperation of T lymphocytes and macrophages, thus the impairment of either function decreased anti-tumor activity.     

Cancer targeting Gene-Viro-Therapy of liver carcinoma by dual-regulated oncolytic adenovirus armed with TRAIL gene

Liver cancer is a common and aggressive malignancy, but available treatment approaches remain suboptimal. Cancer targeting Gene-Viro-Therapy (CTGVT) has shown excellent anti-tumor effects in a preclinical study. CTGVT takes advantage of both gene therapy and virotherapy by incorporating an anti-tumor gene into an oncolytic virus vector. Potent anti-tumor activity is achieved by virus replication and exogenous expression of the anti-tumor gene. A dual-regulated oncolytic adenoviral vector designated Ad·AFP·E1A·E1B (Δ55) (Ad·AFP·D55 for short thereafter) was constructed by replacing the native viral E1A promoter with the simian virus 40 enhancer/α-fetoprotein (AFP) composite promoter (AFPep) based on an E1B-55K-deleted construct, ZD55. Ad·AFP·D55 showed specific replication and cytotoxicity in AFP-positive hepatoma cells. It also showed enhanced safety in normal cells when compared with the mono-regulated vector ZD55. To improve the anti-hepatoma activities of the virus, the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) gene was introduced into Ad·AFP·D55. Ad·AFP·D55-TRAIL exhibited remarkable anti-tumor activities in vitro and in vivo. Treatment with Ad·AFP·D55-TRAIL can induce both autophagy owing to the Ad·AFP·D55 vector and caspase-dependent apoptosis owing to the TRAIL protein. Therefore, Ad·AFP·D55-TRAIL could be a potential anti-hepatoma agent with anti-tumor activities due to AFP-specific replication and TRAIL-induced apoptosis.     

Do anti-tumor necrosis factors induce response and remission in patients with acute refractory Crohn's disease? A systematic meta-analysis of controlled clinical trials

To determine whether anti-tumor necrosis factors induce clinical response and remission in patients with Crohn's disease, PUBMED, OVID, and SCOPUS databases were searched for studies investigated the efficacy of anti-tumor necrosis factors on CD. Data were collected from 1966 to 2005 (up to 31 December). Types of outcome investigated were response (decrease in CDAI score >/=70 points) and remission (CDAI score 相似文献   

Tumor necrosis factor-alpha augments tumor effects in isolated hepatic perfusion with melphalan in a rat sarcoma model

Isolated hepatic perfusion (IHP) is an attractive approach to treating nonresectable liver tumors, because the effects of systemic chemotherapy are poor and its application is hampered by severe general toxicity. In clinical and experimental settings, the efficacy of isolated limb perfusion (ILP) with tumor necrosis factor-alpha (TNF alpha) in combination with melphalan to treat melanoma in transit and soft-tissue sarcoma has been well established. In an ILP model in rats, the authors previously observed synergistic anti-tumor effects of TNF and melphalan on BN 175 soft-tissue sarcoma extremity tumors. The aim of the current study was to determine whether similar synergy in anti-tumor effects could be achieved by treating experimental BN 175 soft-tissue sarcoma liver tumors by IHP using these agents. The authors found that IHP with TNF and melphalan resulted in a dramatic increase in regional concentrations of perfused agents with virtually no concomitant systemic leakage. Isolated hepatic perfusion with only carrier solution resulted in a significantly diminished growth rate of BN 175 liver tumors compared with the growth rate of tumors in nonperfused rats. Perfusion with melphalan alone resulted in minimal anti-tumor effects. Perfusion with only TNF had a slight growth-stimulatory effect on the BN 175 liver tumors, but no negative effects on tumor growth were observed. When TNF was added to melphalan, a dramatic anti-tumor effect was observed. Thus, as in the rat ILP setting, the anti-tumor effect is augmented when TNF is added to IHP with melphalan to treat BN 175 soft-tissue sarcoma tumor-bearing rats. Strikingly, the tumor response was potentiated at relatively low concentrations of TNF compared with concentrations that elicited synergy with melphalan in ILP.     

雷公藤甲素的抗肿瘤活性研究

目的研究雷公藤甲素在体内、体外抗肿瘤活性。方法选取人乳腺癌细胞MCF-7、肝癌细胞SMMC-7721、骨髓白血病细胞(HL-60),不同浓度的雷公藤甲素分别处理3种肿瘤细胞,MTT法分别检测雷公藤甲素的抗肿瘤活性;选取荷MCF-7肿瘤裸鼠,给予雷公藤甲素,通过肿瘤生长抑制率,考察雷公藤甲素的体内抗肿瘤活性。结果雷公藤甲素对3种肿瘤细胞均有抑制作用,呈剂量依赖性,IC50分别为42.1、54.83、1.9 nmol/L,雷公藤甲素可显著抑制肿瘤生长。结论雷公藤甲素体内外实验均显示良好抗肿瘤活性。     

Transcutaneous immunization with cytotoxic T-cell peptide epitopes provides effective antitumor immunity in mice

Much attention has been focused on transcutaneous immunization strategies to stimulate systemic cytotoxic T lymphocyte responses leading to anti-tumor or anti-microbial immunity. Here we report that topical application of vaccines consisting of synthetic peptides formulated with imiquimod, a Toll-like receptor agonist that functions as a potent adjuvant generates strong T cell responses that exhibit effective anti-tumor effects in a murine melanoma model system. These results support the use of peptide-based transcutaneous vaccines as a noninvasive and effective strategy for anti-tumor immunotherapy.     

Peplomycin and liblomycin, a new analogues of bleomycin

Peplomycin, derivative of Bleomycin which was developed because of a lower pulmonary toxicity and broader anti-tumor spectrum in animal study, also showed to be effective in clinical studies on adenocarcinomas such as prostatic cancer, breast cancer, etc. as well as on squamous cell carcinoma and the anti-tumor spectrum were observed to be broader than those of Bleomycin. Further, the combination therapy with other anti-cancer drugs showed higher effectiveness. Clinically lower pulmonary toxicity was also observed. Liblomycin, derivative of Bleomycin having a lipophyllic characteristics, was developed since it showed in animal studies a lower pulmonary toxicity than that of Peplomycin and broader anti-tumor spectrum. At present, this substance is under Phase I study in Japan.     

枸杞多糖增强效应T细胞增殖和杀瘤活性机制的研究

目的观察枸杞多糖对效应T细胞增殖和杀瘤活性机制的作用。方法通过密度梯度离心法分离人外周血单核细胞并培养其为树突状细胞,用冻融法制备肝癌细胞的全抗原并致敏树突状细胞;通过混合淋巴细胞实验,获取树突状细胞激活的特异性效应T细胞。用MTT法测定效应T细胞对肝癌细胞的杀伤率。观察枸杞多糖在树突状细胞的成熟及T细胞的激活、增殖和杀瘤过程中的作用。结果枸杞多糖单独对T细胞无明显的增殖能力,但能促进树突状细胞的成熟并且能增强效应T细胞的增殖能力和杀瘤活性。结论枸杞多糖能促进树突状细胞成熟和增强效应T细胞的增殖和杀伤肿瘤的活性。