Phase I/II trial of PIXY321 (granulocyte-macrophage colony stimulating factor/interleukin-3 fusion protein) for treatment of inherited and acquired marrow failure syndromes

Fourteen paediatric patients with advanced amegakaryocytic thrombocytopenia (AMT) or other bone marrow (BM) failure syndromes were enrolled on one of two phase I/II dose escalation studies of PIXY321. PIXY321 was administered subcutaneously in doses ranging from 250 to 750 mg/m²/d. No dose-limiting toxicity was observed. Peak absolute neutrophil count (ANC) was higher than baseline in all patients. Most transfusion-independent patients demonstrated elevation in haematocrit and/or platelet count. Trilineage haemopoietic responsiveness was evident in the three transfusion-independent patients. In these paediatric populations PIXY321 is well tolerated and merits consideration as a potential therapy.     

人白细胞介素2基因转移表达及抗乙型肝炎病毒和诱导LAK细胞活性的研究

为了探讨白细胞介素（ＩＬ）－２转基因表达抗乙型肝炎病毒（ＨＢＶ）的作用，应用逆转录病毒载体ｐＺＩＰＳｖ（Ｘ）－包装细胞系ＰＡ３１７基因转移系统，研究了人ＩＬ－２ｃＤＮＡ转移和表达，以及抗ＨＢＶ和诱导淋巴因子激活的杀伤细胞（ＬＡＫ细胞）的作用。所构建的人ＩＬ－２重组表达载体ｐＺＩＰｈｕＩＬ－２，以ＤＮＡ－磷酸钙共沉淀技术转染ＰＡ３１７细胞，筛选到Ｇ４１８抗性克隆，分泌假病毒颗粒达１０６ＣＦＵ／ｍｌ。以假病毒颗粒感染２．２．１５细胞系及正常人外周血单个核细胞，分泌ＩＬ－２水平可达６ＩＵ／ｍｌ，明显抑制２．２．１５细胞ＨＢｓＡｇ的分泌表达，与１００ＩＵ／ｍｌ重组的ＩＬ－２一样可诱导出相似的ＬＡＫ细胞活性。而不含ＩＬ－２ｃＤＮＡ的ｐＺＩＰＳＶ（Ｘ）的假病毒颗粒则无此作用。提示逆转录病毒载体。包装细胞系可以成功地实现人ＩＬ－２ｃＤＮＡ的转移和低水平的分泌和表达，并可明显抑制２．２．１５细胞分泌ＨＢｓＡｇ及诱导ＬＡＫ细胞活性。     

Th—LAK细胞对肺癌试验治疗的观察

利用Th-LAK细胞对46例肺癌患者进行了临床试验治疗的观察。结果表明,71.7%的病例原有症状和体征都有不同程度的改善;各期综合的一年生存率为82.6%,高于其它疗法的生存率;部分病例的肿瘤及转移淋巴结有不同程度的缩小。     

MTT比色法的改良及其在LAK细胞活性检测中的应用

目的：改良四甲基偶氮唑盐（MTT）比色法,并用来检测淋巴因子激化的杀伤细胞（LAK细胞）活性。方法：在MTT代谢产物甲瓒DMSO溶解后的最佳吸光度测定及效靶细胞不同孵育时间的比较等方面进行研究。结果：MTT代谢产物甲瓒DMSO溶解后的最佳吸光度为510nm,效靶细胞最佳孵育时间为4h。结论：改良的MTT比色法优于常规的MTT比色法。     

淋巴因子激活的杀伤细胞与自发性高血压大鼠的一氧化氮样内皮依赖性舒血管因子

目的探讨淋巴因子激活的杀伤细胞（ＬＡＫ细胞）与一氧化氮样内皮依赖性舒血管因子（ＥＤＲＦ）作用的相关性。方法动态观察自发性高血压大鼠（ＳＨＲ，ｎ＝５）ＬＡＫ细胞的增殖、活性及其所表达的ＳＯＤ样物质活性当量、自由基清除效应。同时观察胸主动脉环对乙酰胆碱（Ａｃｈ）的舒张反应，以及上述ＬＡＫ细胞与ＳＯＤ标准品对主动脉环（ｎ＝３０）的Ａｃｈ舒张反应的影响，与等量ＷＫＹ大鼠比较。结果与ＷＫＹ比，ＳＨＲ的ＬＡＫ细胞增殖、活性及其所表达的ＳＯＤ样物质和其自由基清除效应均明显降低（Ｐ＜０．０５），经ＬＡＫ细胞（无论来自ＷＫＹ还是ＳＨＲ）孵育后的主动脉环对Ａｃｈ的舒张反应均有不同程度增加。标准ＳＯＤ与ＬＡＫ细胞有相同效应。结论ＬＡＫ细胞可能通过表达ＳＯＤ样物质来减少ＮＯ氧化分解从而增强主动脉环的ＮＯ样舒血管作用。     

淋巴因子激活的LAK／TIL细胞的表型

为研究肿瘤生物疗法中淋巴因子激活的杀伤细胞及其功能和特征与表型之间的关系，用流式细胞术（ＦＣＭ）结合抗体双荧光标记法对培养的淋巴因子激活杀伤细胞作连续的多项表型监测，发现存在着ＣＤ３－ＣＤ１６＋ＣＤ５６＋型，ＣＤ４＋型，ＣＤ８＋ＣＤ２８＋型，ＣＤ８＋ＣＤ２８－型的淋巴因子激活杀伤细胞的亚型。研究了ＣＤ３－ＣＤ１６＋ＣＤ５６＋亚型、ＣＤ８＋ＣＤ２８＋亚型的杀伤活性，显示了对不同的肿瘤靶细胞有不同的敏感性。     

An inhibitory factor of DNA synthesis derived from the medium of short-time incubated human lymphocytes

The medium of human tonsillar lymphocytes incubated for 4 h without any in vitro activation was found to contain a non-dialyzable, soluble inhibitor of DNA synthesis. The spontaneous- as well as the PHA-stimulated DNA synthesis of lymphocytes were both affected by the inhibitor, which was not cytotoxic for its target cells. The factor was isolated by ammonium sulphate precipitation and was purified on a DEAE-cellulose column. It is assumed that the factor is continuously produced by lymphocytes in vivo and plays a role in the homeostatic regulation of lymphocytes.     

白细胞介素-Ⅱ孵育自体淋巴细胞回输治疗慢性乙型肝炎及其机理初步探讨

用白细胞介素Ⅱ(IL-Ⅱ)孵育自体淋巴细胞回输治疗慢性乙型肝炎24例,并另以23例作对照。结果表明,治疗组与对照组HBeAg阴转率分别为54.2％和17.4％(P<0.05);HBeAg/抗-HBe转换率分别为37.5％和8.7％(P<0.05);治疗3个月谷丙转氨酶(ALT)复常率分别为83.3％和52.2％(P<0.05)。治疗组HBeAg/抗-HBe转换者均为慢性活动性肝炎,治疗中多伴有血清球蛋白升高;未出现HBeAg/抗-HBe转换者11例中有7例出现一过性ALT升高。     

Tumoricidal activation of murine resident peritoneal macrophages on pancreaticcarcinoma by interleukin-2 and monoclonal antibodies

INTRODUCTION Macrophages play an important role in tumor lysis and growth inhibition. They can be activated to a tumoricidal state by a variety of agents such as IFNr, TNFa or IL2. The killing machanisms of activated macrophages have been extensively investigated[1,2]. Recently, it has been proved that antibody dependent cellular cytotoxicity (ADCC) is one of the potent arms to lyse tumor cells resistant to cytotoxic macrophages,and that the antitumorous effect of a macrophage activator is significantly augmented by the combined use of mAbs capable of inducing ADCC to tumor cells[3].     

Characterization and partial purification of a novel cytotoxic lymphokine (factor 2) produced by a human B cell line (Karpas 160).

A subclone (160b) of the human B cell (Karpas 160) was shown to produce a novel cytotoxic lymphokine [Factor 2 (F2)] in addition to tumour necrosis factor alpha (TNF-alpha) and beta (TNF-beta). F2 was found to have selective toxicity to numerous human tumour cell lines, particularly the erythroleukaemic cell line K562, whereas TNF-alpha/beta were not cytotoxic to these cells, even at relatively high concentrations. Our studies have shown that F2 activity in crude preparations is heterogeneous both in its molecular weight, isoelectric point (pI) and hydrophobicity, which depends not only on the source of F2 cytotoxicity but also on the conditions of methods for its production. Our studies also showed that F2 was separable from TNF by DE52, S-300 gel filtration and Rotofor isoelectric focusing. F2 was partially purified up to 1000-fold by two procedures. The major active form, as assessed by gel filtration was of mol. wt of 45-67 kDa. On SDS-PAGE, F2 activity was recovered mainly from two regions of the gels corresponding to 10-14 kDa and 60-70 kDa. Antibodies of human TNF-alpha, TNF-beta, IFN-alpha, IFN-gamma, and TGF-beta failed to prevent F2-mediated cytotoxicity to K562. F2 activity was not inhibited by mannose-6-PO4 or mouse mAb to rat granule content, both of which have been reported to block human natural killer cytotoxic factor. Our studies indicated that F2 is likely to be a distinct human cytokine with selective cytotoxic activity against tumour cells.