旋毛虫抗小鼠体内SP2/0肿瘤作用的研究

目的观察旋毛虫感染对BAlB/c小鼠体内SP2/0瘤细胞生长的抑制作用。方法用不同剂量、不同方法致弱的旋毛虫感染BALB/c小鼠,在不同时间接种SP2/0细胞,于荷瘤后20 d解剖小鼠,测定小鼠体内肿瘤的体积、重量和无瘤生长小鼠比率,检测T淋巴细胞亚群。结果1)未致弱组、紫外线致弱组和60Co致弱组小鼠肿瘤体积和重量均显著低于对照组(P<0.01),CD4+/CD8+显著高于对照组(P<0.05);无瘤生长小鼠比率显著高于对照组;2)接种750条、500条和250条旋毛虫组肿瘤体积和重量均显著低于对照组(P<0.01),CD4+/CD8+显著高于对照组(P<0.05),无瘤生长小鼠比率显著高于对照组;3)接种3 d1、1 d和21 d后荷瘤组的肿瘤体积和重量均显著低于对照组(P<0.01),CD4+/CD8+显著高于对照组(P<0.05),无瘤生长小鼠比率显著高于对照组。结论未致弱的旋毛虫和经不同方法致弱的旋毛虫对BALB/c小鼠体内的SP2/0骨髓瘤细胞的生长均有抑制作用,接种剂量增加,抑瘤效果越明显,但对实验动物的机体损伤也加重。接种11 d后荷瘤组的抑瘤效果好于接种3 d后荷瘤组和21d后荷瘤组。     

甲醛固定的H22细胞联合IL-2抑制小鼠H22肝癌生长效果观察

目的 观察甲醛固定的H22肿瘤细胞联合IL-2对小鼠H22肝癌细胞腹部移植瘤生长的影响.方法 20只小鼠,均采用股部皮下1×105个H22细胞的方法建立小鼠H22肝癌皮下移植瘤模型.将其随机分为A、B、C、D组,各5只.肿瘤直径超过1 cm后A、B、C、D组分别瘤内注射生理盐水、甲醛固定H22细胞、IL-2、甲醛固定H22细胞+IL-2各0.5 mL.H22细胞浓度为106/mL,IL-2浓度为8×103 IU/mL.每3d注射1次,共注射4次.治疗结束处死动物,取瘤称重,并计算抑瘤率.采用免疫组化SABC法对肿瘤组织染色,光景下观察瘤内CD8+T淋巴细胞表达情况.结果 A、B、C、D组肿瘤治疗后质量分别为(6.683 ±1.102)g、(2.851 ±0.223)g、(2.282 ±0.443)g、(1.052±0.225)g,B、C、D组与A组相比明显降低,D组与B、C组相比明显降低(P均＜0.05).B、C、D组抑瘤率分别为51.58％、60.54％、80.96％.D组小鼠瘤体内大量肿瘤细胞变性和坏死,且瘤内有大量CDs+T淋巴细胞浸润,阳性产物主要呈现在细胞膜上并被染成棕褐色.B、C组内仅见少量CD8+T淋巴细胞.结论 甲醛固定的H22细胞联合IL-2具有延迟荷瘤小鼠H22肝癌形成,限制生长的作用.     

胸腺法新对晚期非小细胞肺癌化疗患者近远期疗效的影响

目的探讨胸腺法新对晚期非小细胞肺癌化疗患者近远期疗效的影响。方法 60例晚期非小细胞肺癌患者按照随机数表法分为观察组与对照组各30例。对照组采用多西他赛联合顺铂(DP)化疗,观察组在对照组基础上皮下注射胸腺法新,两组连续化疗3个周期。比较两组近期疗效、生活质量改善率,治疗前后外周血T淋巴细胞亚群变化及随访1、2和3年的生存率。结果观察组总有效率(70.00%)显著高于对照组(43.33%,P0.05)。观察组生活质量提高率(70.00%)显著高于对照组(40.00%,P0.05)。观察组治疗后CD3~+、CD4~+、CD4~+/CD8~+较治疗前显著增加(P0.05);对照组治疗后CD3~+、CD4~+、CD4~+/CD8~+较治疗前降低(P0.05);观察组治疗后CD3~+、CD4~+、CD4~+/CD8~+显著高于对照组(P0.05)。两组随访1年生存率比较无统计学差异(P0.05);观察组随访2年和3年的生存率显著高于对照组(P0.05)。结论胸腺法新对晚期非小细胞肺癌化疗患者近远期疗效显著,可改善患者预后,提高生存率。     

老年肺癌患者淋巴细胞亚群、白细胞介素-6、肿瘤坏死因子-α在外周血中的表达及意义

目的探讨老年肺癌患者淋巴细胞亚群、白细胞介素(IL)-6、肿瘤坏死因子(TNF)-α在外周血中的表达及意义。方法流式细胞仪检测老年肺癌患者抗凝全血中淋巴细胞亚群的表达;通过酶联免疫吸附法(ELISA)检测血清IL-6、TNF-α的水平,并分别与正常老年人结果进行对比。结果小细胞肺癌组与非小细胞肺癌组CD3~+、CD3~+CD4~+、CD4~+/CD8~+和自然杀伤(NK)细胞表达水平均明显低于正常组(P<0.05);CD3~+CD8~+、CD3~-CD19~+略高于正常组,但无统计学差异(P>0.05)。与正常组比较,小细胞肺癌组与非小细胞肺癌组血清IL-6、TNF-α水平均明显高于正常组(P<0.05)。Ⅲ~Ⅳ期组CD3~+、CD3~+CD4~+、CD4~+/CD8~+、CD3~-CD19~+和NK细胞数均低于Ⅰ~Ⅱ期组,且CD3~+CD8~+细胞数高于Ⅰ~Ⅱ期组,但差异无统计学意义(P>0.05),Ⅲ~Ⅳ期组血清IL-6、TNF-α水平明显高于Ⅰ~Ⅱ期组(P<0.05)。结论老年肺癌患者存在免疫功能及细胞因子网络紊乱,共同参与了肺癌的发病机制。     

加味参芪汤对老年肺癌患者并发症的改善作用

目的探讨加味参芪汤对老年肺癌化疗患者并发症的改善作用。方法将该院2010年1月至2013年1月接诊的115例老年肺癌患者,随机分为治疗组(63例)和对照组(52例),对照组采用TP(紫杉醇+卡铂)方案化疗,治疗组在对照组方案的基础上服用加味参芪汤治疗。观察患者治疗前后肿瘤坏死因子(TNF)-α、白细胞介素(IL)-2、IL-4、IL-10、T细胞亚群和自然杀伤(NK)细胞水平的变化,并进行肿瘤患者体力状况(KPS评分)、体重变化和毒副反应评估。结果 4个化疗周期后对照组患者的TNF-α、IL-2、CD3+、CD4+、CD4+/CD8+和NK细胞水平明显下降,IL-4和IL-10水平则显著上升;治疗组TNF-α、IL-2、IL-10、CD3+、CD4+、CD4+/CD8+和NK细胞水平与治疗前并无明显差异,而IL-4水平明显降低(P<0.05)。治疗组与对照组相比,治疗组的KPS评分、体重变化和毒副反应评估要明显优于对照组(P<0.05)。结论加味参芪汤对老年肺癌化疗患者的并发症具有明显的改善作用。     

光敏剂喜泊分对H22肝癌小鼠移植瘤的放射增敏效果

目的观察光敏剂喜泊分对H22肝癌小鼠移植瘤的放射增敏效果。方法建立H22肝癌细胞小鼠移植瘤模型,将荷瘤小鼠随机分为对照组、放疗1组、联合治疗1组、放疗2组、联合治疗2组、放疗3组、联合治疗3组,观察第14天瘤体积、抑瘤率、增敏系数、小鼠生存时间、病理、细胞凋亡等指标。结果喜泊分联合放疗能够抑制小鼠移植瘤生长,尤其联合治疗2组,第14天瘤体积明显减小、增敏系数1,有最佳增敏效果,能够明显延长小鼠生存时间,病理HE染色显示,联合治疗2组肿瘤细胞坏死数量最多,坏死范围最大,流式细胞仪检测细胞凋亡显示联合治疗2组凋亡率最高。结论喜泊分联合放疗可以有效减小瘤体积,提高抑瘤率,延长小鼠生存时间,促使细胞凋亡,具有良好的放射增敏效果。     

低剂量^131I标记的抗癌胚抗原抗体C50联合化疗诱导裸鼠结直肠癌移植瘤细胞凋亡

目的从诱导细胞凋亡的角度,探讨低剂量131Ⅰ标记的抗癌胚抗原(CEA)单抗C50(131Ⅰ-C50)对结直肠癌移植瘤的治疗作用及联合5-氟尿嘧啶(5-FU)化疗的效果.方法建立表达CEA的人LoVo结直肠癌荷瘤裸鼠模型.接种第9天,分别采用5-FU、75μ131I-C50及5-FU联合131IC50尾静脉注射治疗裸鼠移植瘤.接种第15天,肿瘤组织进行HE染色和末端脱氧核糖核酸转移酶介导的荧光素-dUTP缺口末端标记法(TUNEL法)凋亡细胞染色,计算肿瘤细胞凋亡指数.结果光学显微镜下各组裸鼠移植瘤组织未见细胞溶解、坏死表现.TUNEL法染色示空白对照组、化疗组、放射免疫治疗(RAIT)组及RAIT+化疗组的凋亡指数分别为(0.29±0.08)%,(18.68±2.69)%,(40.88±4.54)%和(62.33±8.00)%,各组两两之间凋亡指数差异均有显著性(P值均＜0.001).结论低剂量RAIT可通过诱导肿瘤细胞凋亡达到治疗结直肠癌的目的,RAIT联合5-FU化疗可明显增强诱导肿瘤细胞凋亡的效果.     

环孢素A对感染旋毛虫小鼠的免疫调节作用

目的观察环孢素A(CsA)对感染旋毛虫小鼠免疫功能的影响。方法小鼠腹腔注射环孢素A感染旋毛虫后,分别采用ELISA和流式细胞仪检测小鼠不同时期外周血中IL-2含量、CD4+及CD8+T淋巴细胞的百分率。结果(1)单纯感染旋毛虫小鼠感染后1~5周IL-2的含量较正常组明显增加;注射CsA组小鼠感染后1周IL-2的含量较单纯感染组明显降低,感染后2~4周IL-2的含量与单纯感染组相比无明显差异,但仍高于正常组。(2)单纯感染旋毛虫小鼠感染后1~4周,CD4+T淋巴细胞较正常组明显减少,CD8+T淋巴细胞明显增多,CD4+/CD8+比值明显下降;注射CsA组小鼠CD4+T淋巴细胞细胞较单纯感染组明显减少,而CD8+T淋巴细胞无明显变化。结论CsA对感染旋毛虫小鼠具有一定的免疫调节作用。     

参芪扶正注射液对肺癌化疗患者造血功能和免疫功能的影响

目的探讨参芪扶正注射液用于肺癌化疗患者的疗效。方法将78例中晚期非小细胞肺癌患者采用双盲随机对照原则,按入院先后顺序分成观察组30例、对照组23例及安慰剂组25例。三组均予TP方案化疗,化疗前3 d观察组予参芪扶正注射液250 mL/d,对照组予参麦注射液50 mL/d,安慰剂组予0.9%氯化钠250 mL/d,共24 d。检测各组治疗前及化疗结束时血常规、T淋巴细胞亚群(CD3+、CD4+、CD8+、CD4+/CD8+)和NK细胞比例。结果观察组白细胞和血红蛋白降低程度明显轻于安慰剂组(P<0.05),CD3+、CD4+、CD4+/CD8+水平均明显高于安慰剂组(P<0.05),NK细胞三组比较差异无统计学意义。结论参芪扶正注射液能减轻化疗对肺癌患者骨髓造血功能的抑制,并能促进化疗患者骨髓造血功能的恢复;提高化疗患者的细胞免疫功能。     

低剂量~(131)I标记的抗癌胚抗原抗体C50联合化疗诱导裸鼠结直肠癌移植瘤细胞凋亡

目的　从诱导细胞凋亡的角度 ,探讨低剂量13 1I标记的抗癌胚抗原 (CEA)单抗C5 0 (13 1I C5 0 )对结直肠癌移植瘤的治疗作用及联合 5 氟尿嘧啶 (5 FU)化疗的效果。方法　建立表达CEA的人LoVo结直肠癌荷瘤裸鼠模型。接种第 9天 ,分别采用 5 FU、75 μCi13 1I C5 0及 5 FU联合13 1I C5 0尾静脉注射治疗裸鼠移植瘤。接种第 15天 ,肿瘤组织进行HE染色和末端脱氧核糖核酸转移酶介导的荧光素 dUTP缺口末端标记法 (TUNEL法 )凋亡细胞染色 ,计算肿瘤细胞凋亡指数。结果　光学显微镜下各组裸鼠移植瘤组织未见细胞溶解、坏死表现。TUNEL法染色示空白对照组、化疗组、放射免疫治疗(RAIT)组及RAIT +化疗组的凋亡指数分别为 (0 .2 9± 0 .0 8) % ,(18.6 8± 2 .6 9) % ,(4 0 .88± 4.5 4) %和(6 2 .33± 8.0 0 ) % ,各组两两之间凋亡指数差异均有显著性 (P值均 <0 .0 0 1)。结论　低剂量RAIT可通过诱导肿瘤细胞凋亡达到治疗结直肠癌的目的 ,RAIT联合 5 FU化疗可明显增强诱导肿瘤细胞凋亡的效果     

Vaccination with IL-18 gene-modified, superantigen-coated tumor cells elicits potent antitumor immune response

OBJECTIVE: To investigate the induction of antitumor immune response by vaccination with interleukin-18 (IL-18) gene-modified, C215Fab-SEA-coated tumor cells. MATERIALS: A B16-C215 cell clone stably expressing C215 antigen was established by transfecting the gene-encoding C215 antigen into B16 melanoma cells. The manipulated tumor cell vaccine was prepared with B16-C215 cells genetically modified with the IL-18 gene, coated with the fusion protein of SEA and the Fab region of C215 mAb (C215Fab-SEA) which specifically binds to the C215 antigen and then irradiated. C57BL/6 mice were vaccinated with IL-18 gene-modified, C215Fab-SEA-coated B16-C215 cells followed by tumor challenge. Tumor growth and survival time were observed. The expansion of CD4+, CD8+ cells in lymphocytes derived from draining lymph node was detected by FACS. Induction of CTL activity by vaccination was measured by 51Cr release assay. RESULTS: IL-18 gene-modified, C215Fab-SEA-coated B16-C215 cell vaccine effectively stimulated lymphocyte proliferation and CD4+, CD8+ cell expansion in vitro. It was more immunogenic than B16-C215 cells genetically modified with IL-18 gene alone or B16-C215 cells coated with C215Fab-SEA alone. Immunization of the mice with the manipulated vaccine elicited protective immunity against the following tumor challenge of parental B16-C215 and wild-type B16 cells. Significant expansion of CD4+, CD8+ T cells was observed in the draining lymph node of the immunized mice when compared with that in unvaccinated mice. Higher CTL activity was induced in vaccinated mice than that in unvaccinated mice. CONCLUSION: Vaccination with IL-18 gene-modified, C215Fab-SEA-coated tumor cells elicited potent antitumor response through induction of tumor-specific immune response.     

Adoptive transfer of gene-engineered CD4+ helper T cells induces potent primary and secondary tumor rejection

Because CD4+ T cells play a key role in aiding cellular immune responses, we wanted to assess whether increasing numbers of gene-engineered antigen-restricted CD4+ T cells could enhance an antitumor response mediated by similarly gene-engineered CD8+ T cells. In this study, we have used retroviral transduction to generate erbB2-reactive mouse T-cell populations composed of various proportions of CD4+ and CD8+ cells and then determined the antitumor reactivity of these mixtures. Gene-modified CD4+ and CD8+ T cells were shown to specifically secrete Tc1 (T cytotoxic-1) or Tc2 cytokines, proliferate, and lyse erbB2+ tumor targets following antigen ligation in vitro. In adoptive transfer experiments using severe combined immunodeficient (scid) mice, we demonstrated that injection of equivalent numbers of antigen-specific engineered CD8+ and CD4+ T cells led to significant improvement in survival of mice bearing established lung metastases compared with transfer of unfractionated (largely CD8+) engineered T cells. Transferred CD4+ T cells had to be antigen-specific (not just activated) and secrete interferon gamma (IFN-gamma) to potentiate the antitumor effect. Importantly, antitumor responses in these mice correlated with localization and persistence of gene-engineered T cells at the tumor site. Strikingly, mice that survived primary tumor challenge could reject a subsequent rechallenge. Overall, this study has highlighted the therapeutic potential of using combined transfer of antigen-specific gene-modified CD8+ and CD4+ T cells to significantly enhance T-cell adoptive transfer strategies for cancer therapy.     

结核病小鼠T淋巴细胞亚群及其表达的四项细胞因子分析

目的 探讨T淋巴细胞(简称T细胞)表达穿孔素(PFN)、颗粒酶B(GzmB)、γ-干扰素(IFN-γ)、白细胞介素-2(IL-2)与结核免疫的关系.方法 60只MK小鼠按随机数字表法分成结核病组和健康对照组,每组30只.以CD3PerCP、CD4FTTC、CD8APC单抗标记T细胞亚群,以藻红蛋白标记PFN、GzmB、IFN-γ、IL-2单抗标记细胞因子,以流式细胞仪检测分析总的淋巴细胞、CD3+、CD4+、CD8+、CD4+CD8+;双阳(DP)T细胞计数和各亚群占淋巴细胞百分率,观察各T细胞亚群内表达PFN、GzmB、IFN-γ、IL-2的阳性细胞计数和各自占淋巴细胞百分率.采用t检验进行统计学分析.结果 (1)CD4+CD8+、DP T细胞均能不同程度地表达PFN、GzmB、IFN-γ、IL-2.PFN、GzmB的表达以CD8+T细胞占优势.IFN-γ、IL-2的表达以CD4+T细胞占优势.(2)T细胞亚群细胞计数结果与T细胞亚群占总淋巴细胞百分率结果,所反映的T细胞免疫变化趋势可能相似,也可能不同甚至相反.(3)两组间表达PFN的T细胞差别不明显,但结核病组表达GzmB的各个T细胞亚群计数和CD3+%、CD8+%、DP%高于对照组(t值为-3.72～4.13.均P<0.05).(4)结核病组表达IFN-γ的CD3+、CD4+T细胞计数高于对照组;但结核病组表达IL-2的CD8+、DP T细胞计数和CD3+%、CD4+%、CD8+%、DP%均低于对照组(t值为2.62～3.46,均P<0.05).结论 、CD,4+、CD8+、DPT细胞均能不同程度地表达PFN、GzmB、IFN-γ、IL-2;联合评价T细胞各亚群计数和其占淋巴细胞百分率更能判断免疫学状态.     

The age-dependent cytokine production by murine CD8+ T cells as determined by four-color flow cytometry analysis

Flow cytometry has been used to simultaneously examine intracellular cytokine production and expression of CD44 and CD45RB by murine CD8+ T cells derived from young (2-3 months) or aged (18-22 months) mice. Cytokine production by aged CD8+ T cells differs from that by CD8+ T cells derived from young animals in that a significantly higher percentage of the aged can be triggered to produce interleukin (IL)-4, interferon (IFN)-gamma, and tumor necrosis factor alpha (TNF alpha). Conversely, a greater fraction of young CD8+ T cells produce IL-2. Aged mice not only have a higher percentage of CD8+/CD44hi T cells, but also a larger fraction of these cells are IFN-gamma+ and IL-4+, while a lower fraction are IL-2+ than is observed in young CD8+/CD44hi T cells. In terms of relative contribution to total cytokine synthesis, a greater fraction of CD8+ T cells produce IFN-gamma and IL-4 than in CD4+ T cells, whereas CD4+ T cells are the major producers of IL-2.     

特发性血小板减少性紫癜患者CD3^＋、CD4^＋和CD8^＋T淋巴细胞凋亡率的研究

目的:通过检测CD3+、CD4+、CD8+T淋巴细胞的凋亡率,探讨T淋巴细胞凋亡在特发性血小板减少性紫癜(ITP)免疫发病机制中的作用。方法:应用流式细胞仪检测ITP患者外周血CD3+、CD4+、CD8+T淋巴细胞的凋亡率;分离ITP患者和正常人外周血单个核细胞(PBMC),分成A、B 2组,A组加入白介素2(IL-2),B组加入IL-2和地塞米松共培养,分别于24和48 h收获细胞,应用流式细胞仪检测CD3+、CD4+、CD8+T淋巴细胞的凋亡率。结果:①ITP患者组CD3+、CD4+、CD8+T淋巴细胞凋亡率均明显低于正常对照(均P<0.01);②细胞培养24 h时ITP患者组A、B 2组间CD3+、CD4+、CD8+T淋巴细胞凋亡率间均差异无统计学意义(均P>0.05),而48 h时B组CD3+、CD4+、CD8+T淋巴细胞的凋亡率均明显高于A组(P<0.05或0.01);正常对照组24和48 h B组CD3+、CD4+、CD8+T淋巴细胞凋亡率均明显高于A组(均P<0.05);③ITP患者组细胞培养24 h后A、B 2组间CD3+、CD4+、CD8+T淋巴细胞凋亡率的差值均明显低于正常对照组(P<0....     

选择性COX-2抑制剂联合顺铂对肺癌新生血管的影响及机制研究

目的研究选择性环氧化酶2(COX-2)抑制剂尼美舒利(NIM)单独应用以及联合顺铂(DDP)对肺癌A549细胞裸鼠移植瘤血管生成的影响及其机制,为肺癌的治疗提供新思路。方法培养肺癌A549细胞,构建肺癌A549裸鼠移植瘤模型。依据随机数字表法将研究对象分为4组进行药物干预:对照组、NIM组、DDP组、NIM+DDP组,每组8只,观察一般状态。给药后第22天处死裸鼠,获取肿瘤组织,测量称取瘤重、瘤径,计算并比较各组体积抑瘤率。用CD31标记微血管,采用微血管计数法检测各组微血管密度。采用免疫组织化学检测肺癌A549裸鼠移植瘤内血管内皮生长因子的表达。结果本研究成功构建肺癌A549裸鼠移植瘤模型。NIM+DDP组第9、13、17、21天的移植瘤体积增长较其他3组慢(P值均<0.001)。NIM+DDP组体积抑瘤率最大。NIM+DDP组较其他3组移植瘤微血管密度低(F=27.861,P<0.001)。结论选择性COX-2抑制剂单独使用有抑制新生血管生成的作用,推测选择性COX-2抑制剂具有抗肿瘤能力,可能成为肺癌靶向治疗因子。选择性COX-2抑制剂联合DDP对新生血管因子表达的抑制效果更显著,对肺癌A549细胞裸鼠移植瘤的生长的抑制更显著。     

Interleukin-15 combined with an anti-CD40 antibody provides enhanced therapeutic efficacy for murine models of colon cancer

IL-15 has potential as an immunotherapeutic agent for cancer treatment because it is a critical factor for the proliferation and activation of natural killer (NK) and CD8⁺ T cells. Administration of anti-CD40 antibodies has shown anti-tumor effects in vivo through a variety of mechanisms. Furthermore, activation of CD40 led to increased expression of IL-15 receptor-α by dendritic cells, an action that is critical for trans-presentation of IL-15 to NK and CD8⁺ T cells. In this study, we investigated the therapeutic efficacy of the combination regimen of murine IL-15 (mIL-15) with an agonistic anti-CD40 antibody (FGK4.5) in murine lung metastasis models involving CT26 and MC38, which are murine colon cancer cell lines syngeneic to BALB/c and C57BL/6 mice, respectively. Treatment with mIL-15 or the anti-CD40 antibody alone significantly prolonged survival of both CT26 and MC38 tumor-bearing mice compared with the mice in the PBS solution control group (P < 0.01). Furthermore, combination therapy with both mIL-15 and the anti-CD40 antibody provided greater therapeutic efficacy as demonstrated by prolonged survival of the mice compared with either mIL-15 or the anti-CD40 antibody-alone groups (P < 0.001). We found that NK cells isolated from the mice that received the combination regimen expressed increased levels of intracellular granzyme B and showed stronger cytotoxic activity on the target cells. The findings from this study provide the scientific basis for clinical trials using the combination regimen of IL-15 with an anti-CD40 antibody for the treatment of patients with cancer.     

Myeloid differentiation and maturation of SCF+IL-3+IL-11 expanded AC133+/CD34+ cells selected from high-risk breast cancer patients

The AC133 antigen is selectively expressed on subset of CD 34+ cells isolated from leukapheresis products from high risk breast cancer patients receiving chemotherapy plus G-CSF. MiniMACS AC133+ isolated cells contained a mean of 85% (80-90) AC133+ cells. Enriched AC133+ cells coexpressed 80% CD34+, 6.6% CD33+ and 2% CD15+. Separated AC133+ cells contained 600 GFU-GM/10(4) cells and 70 BFU-E/10(4) cells. Flow-cytometric analysis indicated that AC133+ cells were isolated from cells population with low granularity (SS), while CD33+ a CD15+ cells had a high granularity. After a seven-day ex vivo expansion in the presence of SCF + IL-3 + IL11, the expansion of cells increased 19.4 times. The mean percentage of blasts decreased from 100% at the start of culture to 81% on day 3 and 30% on day 7. Promyelocytes were slow to appear with 10% present on day 3, but thereafter increased to 33% on day 7. The appearance of myelocytes and metamyelocytes lagged 3 days behind promyelocytes and continued to increase during culture to become the predominant (30%) cell type on day 7. Very few neutrophils (2%) were observed in any of the cultures on day 7. Monocytes or macrophages were not detected on day 7. By day 7 megakaryocytes were present at low levels (10%). The mean value of CFU-GM in the culture after day 7 of ex vivo expansion in the presence of SCF+IL-3+IL-11 had increased 45-fold, BFU-E 5-fold. After 7 days of expansion with IL-3+SCF+IL-11 cells expressed a mean of 12% CD34+, 8% AC133+, 59% CD33+ and 30% CD15+. The aim of this experiment was to determine whether ex vivo culture of peripheral blood AC133+ cells could generate sufficient numbers of progenitors to potentially abrogate cytopenia after transplantation and passive purging of tumor cells.     

Generation of effector cells from primary lung cancer patients by stimulation with anti-CD3 monoclonal antibody followed by culture with recombinant interleukin-2]

Peripheral blood mononuclear cells obtained from 24 primary lung cancer patients were stimulated with anti-CD3 monoclonal antibody (alpha CD3MoAb) followed by culture with recombinant interleukin-2. The optimal concentration of alpha CD3MoAb for stimulation was 50 ng/ml in the liquid phase, and the sensitization culture was commenced at a cellular concentration of 1 x 10(6)/ml. Patients entered into this study were 14 cases of adenoca rcinoma, 7 of squamous cell carcinoma, and 3 of small cell carcinoma. After 4-6 days of stimulation with alpha CD3MoAb followed by culture with RIL-2 for 5-7 days, the cellular expansion was 3.7 folds (mean). Surface marker analysis of the cells revealed significant increments of CD3+, CD8+, HLA-DR+, and IL-2R+ cells after sensitization culture. In 2 cases, fresh autologous tumor cells could be obtained from surgical specimens. Effector cells generated in those 2 cases did not show significant cytotoxic activity against autologous tumor cells in 4 hr 51Cr release assay. In 5 cases, cytotoxicity against established lung cancer cell lines, STC-1 and L0301, were analyzed. In all cases, effector cells showed significant cytolytic activity against both targets. The sensitization culture utilizing alpha CD3MoAb was easy to perform and feasible for the majority of patients, and it is considered that utilization of this culture system would be worth while for adoptive immunotherapy in primary lung cancer patients.     

Effect of Fuzheng Yiliu decoction combined with chemotherapy on patients with intermediate and late stage gastrointestinal cancer

AIM: To investigate the therapeutic effects of Fuzheng Yiliu (strengthening the body resistance to inhibit tumor) decoction combined with chemotherapy on the patients with intermediate and late stage gastrointestinal cancer. METHODS: Sixty patients were randomly divided into treatment group (chemotherapy combined with Fuzheng Yiliu decoction) and control group (chemotherapy alone). Four indexes, including the tumor recent remission rate (RR), the change of main symptoms, the toxic and side effects caused by chemotherapy and the change of performance status, were observed in the patients. Peripheral blood contents of CD3+, CD4+, CD8+ cells, CD4+/CD8+ and soluble interleukin-2 receptor (sIL-2R) were tested before and after treatment and the values were compared with those of healthy peoples. RESULTS: The improving rate of main symptoms (69.6%) and performance status (56.7%) were significantly higher in the treatment group than in the control group (34.8%, 26.7%, P<0.05). The occurrence rates of grade II toxic and side-effects on both bone marrow (13.3%) and digestive tract (30%) were lower in the treatment group compared to the control group (36.7%, 63.3%, P<0.05). Before treatment, the proportion of CD3+, CD4+ and CD4+/CD8+ decreased and the proportion of CD8+ and SIL-2R raised markedly both in the control group and treatment group as compared to the healttiy people. After treatment, that increased of CD3+, CD4+, CD4+/CD8+ increased (62.25±10.01% vs 68.31±9.72%, 36.83±10.44% vs 42.6±9.62%, 1.24±0.65 vs l.66±0.85, P<0.05) and the values of CD8+ and sIL-2R decreased obviously (33.06±7.69% vs 29.24±6.25%, 588.23±216.86 U/mL vs 475.87±211.36 U/mL,P<0.05) in the treatment group, whereas these values were opposite in the control group (64.22±6.91% vs60.63±5.75%,35.62±7.49% vs31.53±5.53%, 32.95±8.28% vs 37.14±7.48%, 1.17±0.43 vs 0.94±0.43, 573.63±214.32 U/mL vs 692.17?21.33 U/mL, P<0.05). CONCLUSION: Fuzheng Yiliu decoction can enhance therapeutic effects of chemotherapy on malignant gastrointestinal tumor, and also reduce the toxic and side effects on bone marrow and digestive tract, thereby improving the quality of life and cellular immunity in patients with malignant gastrointestinal tumor.