人宫颈癌免疫学

近年来,对人宫颈癌抗原、标记物、癌细胞中病毒抗原、患者免疫功能、免疫诊断和免疫治疗等作了广泛研究。 癌相关抗原(TAA) 人宫颈癌抗原性与正常宫颈上皮不同。兔抗宫颈癌抗血清在免疫扩散中与宫颈癌匀浆产生沉淀线,与正常宫颈匀浆无反应,反应有一定特异性。同时癌患     

人到老年

人到老年,会突然醒悟: 生命是有尽头的. 这种感悟会使他们行动起来, 去做一些他们很想做但以前总也没有做的事情, 有时,甚至用他们"意料不到"的方式. 人到老年,才真正认识自己: 什么才真正属于自己, 用一种宽容、舒适和诚实的方式接受自己. 人到老年,方才明白: 东奔西走竭力想去改变的不是别的, 恰恰是自己.     

人THANK基因转染人肝癌细胞及生物学特性

[目的]将人THANK基因转染入人肝癌细胞SMMU-7721中稳定表达,检测转染后细胞的生物学特性变化.[方法]将由pMD18-T-THANK质粒上获得的THANK全长cDNA克隆到真核表达载体pcDNA3.1中;用脂质体法将pcDNA3-THANK导入人肝癌细胞株SMMU-7721中,G418筛选克隆,再以RT-PCR及Western blot检测THANK的表达,流式细胞仪及细胞毒杀伤实验分析转染后细胞的生物学特性变化.[结果]THANK在7721细胞中的表达可抑制7721细胞的生长,使肿瘤细胞的生长停滞于S期,且可在体外诱导强烈的细胞毒性T淋巴细胞(CTL)反应,人外周血单个核细胞(PBMC)对THANK-7721细胞的杀伤活性明显增强.THANK的转染与IFN-γ协同可增强7721细胞的凋亡比例.[结论]THANK基因的转染改变了7721细胞的生物特性,在体外诱导PBMC的CTL反应,有可能用于肝癌的免疫治疗.     

治病与治人

相信很多人都听说过一个“治驼背”的笑话：一个驼背问医生，你能否把我的驼背治好啊？医生说，这个简单。驼背问，简单？怎么治啊？医生说，你趴在地上，我在你的驼背上猛踩几脚不就直了吗!     

人精子诱变研究进展

人精子诱变研究进展黄天华汕头大学医学院汕头市５１５０６３１９７８年,以色列学者Ｒｕｄａｋ在夏威夷大学医学院Ｙａｎａｇｉｍａｃｈｉ教授指导下,将本世纪生殖生物学界的一项重大成就“异种体外受精技术”引入遗传学研究领域,首次制备出人精子染色体标本（１）。继...     

可溶性重组人CD40配体抑制人胶质瘤细胞生长

背景和目的：CD40是存在于B细胞,单核细胞,树突状细胞,内皮细胞和多种肿瘤细胞上的肿瘤坏死因子受体超家族成员,已有研究证明用CD40抗体或CD40配体激活CD40可引起人卵巢癌,结肠癌,乳腺癌和肺癌细胞凋亡或死亡,并抑制鼠体内移植瘤生长,本研究拟了解可溶性重组人CD40配体（srhCD40L)对人胶质瘤细胞增殖的影响。方法：用免疫组化法检测胶质瘤标本中CD40的表达,用流式细胞仪检测人胶质瘤细胞中CD40的表达,然后在其培养基中加入srhCD40L孵育,在分别培养72h和24h后检测细胞增殖和凋亡指标。结果：23个胶质瘤标本中有7个和3个胶质瘤细胞系有1个具有CD40表达。有CD40表达的胶质瘤细胞系与srhCD40L共孵育后发现细胞活力从98%降至87%;凋亡和坏死升高了17.4%。结论：在体外用CD40配体激活CD40可抑制有CD40表达的人胶质瘤细胞的生长,提示CD40配体在治疗胶质瘤上有潜在的临床应用价值。     

人IL-18克隆

从人外周血单核细胞中分离总RNA,进行RT-PCR获得人IL-18成熟编码序列cDNA,测序结果显示,所克隆的人IL-18cDNA编码序列与GenBank所报道的序列完全一致.本研究结果为进一步探讨IL-18的生物学活性和特性奠定了基础,同时亦为利用IL-18进行免疫基因治疗打下了良好的基础.     

顾健人院士题词

三代英才展宏图呕心沥血不言苦众志成城创大业只为苍生镇顽疴。     

一个人的坚持

我是一名肿瘤放射治疗科医生,工作这么多年,接触过许许多多、各式各样的癌症病人,一些病人的面容在脑海中已渐渐模糊,而有一些则让我终生难忘,李强（化名）便是其中一位。     

Anti-human lung giant cell cancer (PG) effect of human LAK cellsin vitro and in nude mice

Human LAK cells were prepared by culturing normal human peripheral blood mononuclear cells (PBMC) with or without rIL-2 and assayed for T cell surface markers as well as anti-tumor activity against PGin vitro and in nude mice. Although the percentages of T₃, T₄ and T₈ positive cells in rIL-2-activated cells did not differ significantly from those of control cellsin vitro, the former showed stronger cytotoxicity than control cells to PG tumor cellsin vitro. In vivo, LAK cells completely inhibited the growth of PG tumor in nude mice, whereas PBMC control cells were to be of no effect. The anti-tumor effect of human LAK cells in nude mice may offer a useful model to study the role of human LAK cells against human tumorin vivo.     

RESEARCHES ON THE ANTI-AUTOLOGOUS TUMOR EFFECTS OF HUMAN CANCER-KILLING CELLS IN NUDE MICE

An investigation of human splenic LAK cells killing autologous tumor in vivo has been performed in this study. Briefly, an ovary embryo carcinoma (OEC) removed surgically from patient was transplanted Into nude mice (s. c. ) and cultured in vitro, that would be used as targets. Lymphokine-activated killer (LAK) cells were generated from splenic lymphocytes of the OEC patient, who died two and half months after operation, by co- culture with recombinant human interleukin- 2 ( rIL- 2 ) in vitro. The results from Winn' s test in nude mice suggested that these LAK cells could effectively inhibit the tumorlgenicity of autologous tumor m vitro.     

CIK的体外增殖及体内外杀瘤活性的实验研究

目的:从人骨髓造血前体细胞体外培养扩增树突状细胞(dendritic cells,DCs),测定其表型及T细胞刺激活性.方法:采用Mini-MACS分离技术,从正常人骨髓、脐血分离CD34~ 造血干细胞,体外以重组hGM-CSF,hTNF-α,hIL-3诱导培养2周,流式细胞术检测扩增细胞的表面表型及细胞内IL-12的表达,体外同种混合淋巴细胞反应检测扩增DCs的T细胞刺激活性.结果:从正常人骨髓、脐血分离得到高纯度(>90%)的CD34~ 造血干细胞,经重组hGM-CSF,hTNF-α的共同诱导培养,扩增得到大量DCs,加人hIL-3可以进一步增加DCs产量;FACS检测表明,扩增的DCs表达HLA-DR,CD40,CD54,CD80,CD86分子,细胞内有hIL-12的P35,P40亚基的表达;与外周血单核细胞培养生成的DCs相比,由CD34~ 干细胞扩增的DCs具有更强的激发同种T细胞增殖的能力.结论:人CD34~ 干细胞体外经诱导培养,可以生成大量功能成熟的DCs,从而为进一步开展DCs的基础及临床研究打下了基础.     

人肺癌细胞NHE—1基因片段的克隆及其反义表达载体的构建

目的:动态观察CIK(cytokine induced killer)细胞的体外增殖,体外的细胞毒活性,及通过动物实验研究其体内的抗肿瘤作用.方法:通过提取健康供血者的PBMC,第0天加入γ-IFN,第1天加入IL-2、抗-CD3单抗和IL-1培养CIK细胞;在流式细胞仪上做动态培养物的表型分析;与LAK细胞作对比,分别用MIT法测定其体外细胞毒活性及对S180荷瘤鼠的体内抗肿瘤作用.结果:CIK细胞在培养2周后获得大量增殖,表型分析表明,CIK细胞属异质性细胞群,在培养的过程中,群体的CD3~ CD56~ 细胞大量扩增达1000多倍,是CIK细胞的主要效应细胞;实验证明,CIK细胞的体外细胞毒活性及对S180荷瘤鼠的体内抗肿瘤作用均强于LAK细胞;其较强的体内抗癌活性可能与荷瘤鼠主体内T细胞活化有关.结论:CIK细胞是一种强于LAK细胞的、新型、高效、具有广谱杀瘤活力的免疫活性细胞.     

In vitro and in vivo susceptibility of human leukemic cells to lymphokine activated killer activity

The susceptibility of human myeloid and lymphoid leukemic blasts to the lytic action of recombinant interleukin-2 (rIL-2)-generated lymphokine activated killer (LAK) cells was analyzed. With the exception of the K562 cell line, all 9 leukemic cell lines tested were resistant to the natural killer activity of freshly isolated peripheral blood lymphocytes (PBL) from healthy donors but were susceptible to the lytic action of PBL cultured for 3 days in the presence of rIL-2. Of the 32 primary myeloid and lymphoid acute leukemia samples investigated, the great majority were natural killer cell-resistant but were variably sensitive to LAK effectors. Variations in LAK activity were observed according to the donor of PBL, while little or no difference was documented in the capacity to elicit LAK activity of PBL cultured with 100 or 1,000 U of rIL-2/ml. Pretreatment of the leukemic target cells with neuraminidase did not increase substantially their sensitivity to LAK activity. LAK cells generated from the PBL of patients at the onset of the disease or in complete clinicohematological remission lysed Raji cells as efficiently as normal LAK effectors. Finally, LAK cells were capable of abrogating the tumor growth in nude mice of a human leukemic T cell line. These findings demonstrate the susceptibility in vitro and in vivo of human leukemic blasts to the lytic effect of LAK cells and point to a possible clinical exploitment of this new form of adoptive immunotherapy in the management of acute leukemia.     

CIK细胞体内外抗肝癌细胞作用

目的：研究肝癌患者CIK（cytokine-induced killer) 细胞的体外杀伤自体肝癌原代细胞的细胞毒活性以及正常人CIK细胞在裸鼠体内的抗肿瘤作用。方法：分别分离获得肝癌患者和下人的外周血单个核细胞（PBMC）,加入细胞因子,体外诱导成CIK细胞,用流式细胞仪对细胞作动态表型分析,并与正常人的CIK细胞作对比。用^51Cr释放法,测定肝癌患者的CIK细胞体外杀伤自体肿瘤细胞的细胞毒性活性。在Balb/c裸鼠皮下接种肝癌细胞BEL－7402,观察CIK细胞对荷瘤鼠的抑瘤作用,并与LAK、PBMC细胞相对比。结果：肝癌患者的CIK细胞体外增殖力强,至培养28天时达到最大增值倍数300多,表型分析结果表明,CD^3 CD56^ 双阳性细胞得到了大量的扩增,其含量由原来的0．23％上升到第21天的17．8％。体外实验表明,肝癌患者的CIK细胞杀伤自体原代肝癌细胞的细胞毒性活性明显高于自体的PBMC细胞。裸鼠体内实验表明,肝癌患者的CIK细胞能够显著抑制肿瘤的生长,其抑瘤率可达84．7％,高于LAK细胞的52．8％及PBMC的37．1％（P＜0．05和P＜0．01）。结论：CIK细胞具有较强的体内外抗肝癌细胞活性,有可能应用于临床上肝癌的过继性免疫治疗。     

Potentiation by interleukin 2 of Burkitt's lymphoma therapy with anti-pan B (anti-CD19) monoclonal antibodies in a mouse xenotransplantation model

To study the immunotherapeutic potential of monoclonal antibodies (mAbs) directed against the human pan-B-cell antigen CD19, a xenotransplantation model was developed in which the human Burkitt's cell line Daudi is s.c. transplanted into nude mice. IgG1, IgG2b, and IgG2a isotype variants of the anti-CD19 mAb (CLB-CD19) were tested for their capacity to inhibit the growth of 10 x 10(6) Daudi cells injected s.c. into nude mice. When mAb treatment was started 30 min after the injection of tumor cells, only the IgG2a isotype of CLB-CD19 had a marked antitumor effect in vivo. If treatment with IgG2a anti-CD19 mAb alone was delayed until Day 10 after tumor injection, no therapeutic effect was observed. However, the combination of this delayed mAb treatment with recombinant interleukin 2 (rIL-2) inhibited the growth of the Daudi cells in the nude mice, while treatment with rIL-2 alone was ineffective. The results of in vitro experiments showed that peritoneal exudate cells were able to inhibit the proliferation of Daudi cells in the presence of the IgG2a isotype variant of CLB-CD19 mAb but not in the presence of the other CLB-CD19 mAb isotype variants. Fresh nude mouse spleen cells did not mediate antibody-dependent cellular cytotoxicity against CLB-CD19 mAb-sensitized Daudi cells, irrespective of the isotype used for sensitization. However, preculture of these spleen cells with rIL-2 induced antibody-dependent cellular cytotoxicity against CD19+ target cells sensitized with CLB-CD19 mAb of all isotypes. These results indicate that it is possible to enhance mAb-dependent effector systems in vivo with the lymphokine rIL-2.     

Anti-tumor effects of interleukin-2 and interleukin-1 in mice transplanted with different syngeneic tumors

We have studied the anti-tumor effects of human recombinant IL-2, alone or in association with LAK cells, in mice transplanted subcutaneously (s.c.) with the following syngeneic tumors: highly metastatic Friend leukemia cells (FLC), nonmetastatic FLC, lymphoma RBL-5 cells and HeJ16 fibrosarcoma cells. In these tumor models, peri-tumoral injections of IL-2 were more effective in inhibiting tumor growth than a systemic treatment. Although s.c. IL-2 treatment resulted in marked inhibition of tumor growth in mice injected s.c. with highly metastatic FLC, it was not effective in inhibiting growth of FLC in the liver and spleen. IL-2 therapy was more effective at increasing survival time in mice transplanted with non-metastatic FLC or with RBL-5 cells. In mice transplanted with HeJ16 fibrosarcomas, s.c. IL-2 treatment resulted in highly significant anti-tumor effect and survival of 70% of tumor-injected mice. No general correlation was found between in vitro sensitivity or resistance to the cytolytic activity of LAK cells and the anti-tumor effects observed in vivo. Subcutaneous injection of IL-1 beta in mice transplanted with highly metastatic FLC resulted in a marked increase in survival time and inhibition of metastatic tumor growth in liver and spleen. Combined treatment of IL-1 beta and IL-2 produced a synergistic anti-tumor effect: 60% of mice injected with highly metastatic FLC survived. Combined IL-1/IL-2 treatments exerted no anti-tumor activity either in DBA/2 mice injected with antibody to Thy 1.2 antigen or in nude mice, indicating that T cells play important roles during IL-1/IL-2 therapy. In vitro treatment of FLC with IL-1 beta resulted in a slight inhibition of cell multiplication, whereas even high doses of IL-2 did not affect FLC multiplication. Our results indicate that local combined treatments with IL-1 and IL-2 can induce potent, host-dependent (T cell-mediated) anti-tumor effects against highly malignant tumors.     

ANTITUMOR EFFECTS OF HUMAN IL-15 GENE MODIFIED LUNG CANCER CELL LINE

ＡＮＴＩＴＵＭＯＲＥＦＦＥＣＴＳＯＦＨＵＭＡＮＩＬ－１５ＧＥＮＥＭＯＤＩＦＩＥＤＬＵＮＧＣＡＮＣＥＲＣＥＬＬＬＩＮＥＳｈｅｎＹｏｎｇｑｕａｎ１沈永泉ＣｕｉＬｉａｎｘｉａｎ２崔莲仙ＨｅＷｅｉ１何维ＸｕｅＬｉ１薛莉ＢａＤｅｎｉａｎ１巴德年１Ｉｎｓｔ...     

Functional analysis of cytotoxic cells in patients with acute nonlymphoblastic leukemia in complete remission

In the current study, we investigated the cytotoxic ability of peripheral blood mononuclear cells (PBMC) recovered from patients with acute nonlymphoblastic leukemia (ANLL) in complete remission (CR) against natural killer (NK)-sensitive, NK-resistant, autologous and allogeneic leukemic target cells taken at diagnosis. Our purpose was to define the role played by cytotoxic mechanisms in the control of leukemic cell growth in ANLL. Experiments were carried out at resting conditions and after in vitro activation with recombinant interleukin-2 (rIL-2) and anti-CD3 monoclonal antibody (moAb). At resting conditions, PBMC recovered from ANLL patients displayed a NK function that was not significantly different from controls (mean +/- standard error of the mean [SEM]: 21.9% +/- 3.9% versus control values of 27.5% +/- 2.9%; the P value was not significant [NS]), but they were unable to show cytotoxic activity against autologous and allogeneic leukemic cells. After in vitro boosting with rIL-2, PBMC were able to generate lymphokine activated killer (LAK) cells, as demonstrated by an increased killing of NK-resistant Daudi targets (16.3% +/- 2.7%). Although LAK activity was quantitatively lower than in control subjects (mean +/- SEM: 16.3% +/- 2.7% versus control values of 79.8% +/- 3.1%; P less than 0.001), it still exerted a cytotoxic effect against autologous and allogeneic leukemic cells. Similar results were obtained when anti-CD3 moAb was used as a stimulus in vitro. Our data suggest that nonspecific cytotoxic cells may be triggered to exert an in vitro cytotoxic effect on leukemic cells, which could possibly play a key role in vivo in the control of leukemic cell growth regulation.