丝裂霉素C对人胎脾LAK细胞活性的影响

本文对人胎脾LAK细胞活性及丝裂霉素C(MMC)对LAK活性的影响作了初步研究。结果显示,人胎脾淋巴细胞可诱导产生明显的LAK活性,对K562和Raji瘤细胞的杀伤活性分别为74.4％和69.4％(E:T=50:1)。诱生的LAK细胞经不同浓度MMC处理后,DNA合成受到显著抑制(p<0.O1),但仍可保持较高水平的LAK活性。提示细胞增殖对MMC的抑制作用更为敏感,LAK细胞在效应阶段并非绝对有赖于细胞增殖。     

人胎脾LAK细胞活性发育动力学

本文采用4h~(51)Cr释放试验观察了不同眙龄脾的LAK细胞活性。胚胎发育至16周时,胎脾淋巴细胞经IL-2诱导72～96h后,产生明显的LAK活性,对Raji靶细胞的杀伤率为67.7±2.2％,而16周前(15周)的LAK活性低下(20.2±5.8％)。结果还显示,16周后的胎睥LAK活性与眙龄增长无关,与正常成人水平无显著性差异(P>0.05);胎脾LAK前体细胞的发生可能早于NK细胞。     

重组IL-2激活的胎儿肝脾细胞LAK样活性

本研究证明,胎儿肝脾细胞经重组IL-2作用96小时,可产生显著的LAK样活性,在重组IL-2 1000u/ml,E:T为100:1其杀伤率分别达60%和62%。如果这一体外试验结果在体内应用中得到证实,无疑将给临床获取LAK细胞提供一条新的途径。     

流式细胞仪检测人外周血和脾LAK细胞的研究

流式细胞仪检测人外周血和脾ＬＡＫ细胞的研究陈宝安周振英朱敏生高雪芝金宝翠邵泽叶临床上使用淋巴因子激活的杀伤细胞（ＬＡＫ细胞）治疗恶性肿瘤，取得良好效果，但在疗效判断上，仍缺乏有效客观的手段。流式细胞仪（Ｆｌｏｗｃｙｔｏｍｅｔｒｙ，ＦＣＭ）是一种先进的...     

冬虫夏草对白血病人NK和LAK细胞活性的影响

本文采用１２５ＩＵｄＲ释放法，研究了冬虫夏草水提液对缓解期和活动期白血病人ＮＫ和ＬＡＫ细胞活性的影响。结果表明冬虫夏草水提液能显著增强活动期白血病人ＮＫ细胞杀伤Ｋ５６２肿瘤细胞的能力，但对ＬＡＫ细胞活性有抑制作用，前列腺素Ｅ１能阻断冬虫夏草水提液增强ＮＫ细胞活性。由此可见，冬虫夏草水提液增强ＮＫ细胞活性在其抗肿瘤免疫中有着极其重要的作用。     

人外周血树突状细胞对LAK细胞杀伤活性的影响

从人外周血中分离出树突状细胞，体外观察了其对ＬＡＫ细胞活性的影响。发现：５×１０２～１×１０４／ｍｌ树突状细胞对ＬＡＫ细胞活性起增强作用，而５×１０４～１×１０５／ｍｌ树突状细胞却抑制ＬＡＫ杀伤活性。这说明人外周血树突状细胞对ＬＡＫ细胞活性起双相性调节作用。     

人胎肝提取物对小鼠脾细胞增殖反应的影响

本实验采用不同浓度的饱和硫酸铵分段沉淀入胎儿肝浸液,获得一组可溶性蛋白,称为人胎肝提取物(Fetal liver extract,FLE)。对其免疫学活性的研究发现,FLE没有种属限制性,能抑制C_(57)BL/6小鼠脾细胞对ConA的增殖反应。31～50%硫酸铵饱和度沉淀的FLE具有较高的抑制活性。FLE对小鼠脾细胞无明显细胞毒活性,较高浓度时对脾细胞的生长呈现抑制作用。FLE作用2小时即可明显抑制脾细胞的增殖反应,洗去FLE后,其抑制作用不能逆转。其抑制活性可被L-精氨酸解除。     

人外周血树状突细胞在体外对LAK杀伤活性的影响

本文采用三因素不同水平的析因设计，研究了人外周血树状突细胞在体外对ＬＡＫ抗肿瘤作用的影响。结果显示，３种浓度的树状突细胞均能增强ＬＡＫ的杀伤活性（Ｐ＜０．００１），以中等浓度的树状突细胞作用最为明显。加入ＩＬ－２后，树状突细胞对ＬＡＫ抗肿瘤活性的增强程度进一步提高。     

人LAK细胞抗菌杀瘤活性多肽HLP-2b的分离纯化

目的 :分离纯化人LAK细胞小分子抗菌多肽 ,并检测其抗肿瘤细胞株K5 6 2活性。方法 :采用密度梯度离心法分离人外周血单个核细胞 ,并用IL 2和PHA刺激培养。 5 %乙酸匀浆细胞获得其酸溶性提取物。应用制备性酸性尿素聚丙烯酰胺凝胶电泳技术和反相高效液相色谱技术分离纯化多肽 ,Tricine SDS PAGE鉴定其分子量 ,并运用琼脂糖弥散法、MTT法鉴定其抗菌、抗肿瘤活性。结果 :从人LAK细胞酸溶性提取物中纯化出一多肽 ,分子量为 7 9kD ,命名为HLP 2b ,具有抗金黄色葡萄球菌活性和抗肿瘤细胞株K5 6 2活性。结论 :HLP 2b可能为LAK细胞一新的抗菌抗瘤效应分子。     

不同浓度腐胺对人肝细胞增殖及凋亡的影响

目的探讨不同浓度腐胺对体外培养人正常肝细胞增殖、凋亡的影响。方法将体外培养的人正常肝细胞株LO2分为腐胺组与对照组,不同浓度腐胺组以含腐胺浓度分别为0.25、0.50、1.00、10.00、20.00、40.00、80.00、160.00、320.00、640.00μg/mL完全培养基培养细胞,对照组以不添加腐胺完全培养基培养细胞。对照组与不同浓度腐胺组处理的LO2细胞培养12 h后,再以四唑化合物电子耦联显色法（MTS）、流式细胞技术（FCM）分别测定细胞的增殖活性（用吸光度值表示）与凋亡率,并对两组数据进行相关性分析。结果 MTS结果显示,0.25、0.50、1.00、10.00、20.00μg/mL浓度腐胺组LO2细胞吸光度值均较对照组（0.474±0.022）升高,差异有统计学意义（P〈0.05）,并且1.00μg/mL浓度时达到峰值（0.834±0.012）;80.00μg/mL及以上浓度腐胺组LO2细胞吸光度值较对照组降低,差异有统计学意义（P均〈0.01）;40.00μg/mL腐胺组LO2细胞吸光度值（0.477±0.009）与对照组比较,差异无统计学意义（P〉0.05）。流式细胞技术结果显示,0.25、0.50、1.00、10.00、20.00μg/mL浓度腐胺组LO2细胞凋亡率均较对照组（15.23±1.82）%降低,差异有统计学意义（P〈0.05）,并且1.00μg/mL腐胺组细胞凋亡率达到最低值（2.30±1.00）%;80.00μg/mL及以上浓度腐胺组LO2细胞凋亡率较对照组升高,差异有统计学意义（P均〈0.01）;40.00μg/mL腐胺组LO2细胞凋亡率（16.10±1.45）%与对照组比较,差异无统计学意义（P〉0.05）。相关性分析结果显示,各组浓度腐胺处理LO2细胞其细胞增殖与凋亡率呈负线性相关关系（r=-0.989,P=0.000）。结论低浓度（0.25-20.00μg/mL）腐胺对人正常肝LO2细胞增殖有促进作用,而较高浓度（80.00μg/mL及以上）的腐胺则出现明显的诱导凋亡作用,低浓度腐胺促进细胞增殖可能是通过抑制细胞凋亡而实现。     

人LAK细胞抗菌活性多肽HLP-3P21的分离纯化和鉴定

目的　分离纯化人LAK细胞小分子抗菌多肽 ,鉴定其分子特性 ,检测其抗菌活性。方法　采用密度梯度离心法分离人外周血单个核细胞 ,并用IL 2和PHA刺激培养。 5 %乙酸匀浆细胞获得其酸溶性提取物。应用制备酸性尿素聚丙烯酰胺凝胶电泳技术和反相高效液相色谱技术分离纯化多肽 ,Tricine SDS PAGE鉴定其相对分子质量 (Mr) ,运用琼脂糖弥散法鉴定其抗菌活性 ,采用Edman降解法测定其氮端序列 ,并进行肽质谱分析。结果　从人LAK细胞酸溶性提取物中纯化出一多肽 ,Tricine SDS PAGE表观Mr 为 17× 10 3,命名为HLP 3P2 1(humanlymphocytepeptide) ,具有抗致病性大肠杆菌ML 35P及铜绿假单胞菌ATCC6 5 92 2的活性。其氮端序列为PKRKAEGDAK ,肽质谱的检索分析未发现匹配度大于 5 0 %的蛋白质分子。结论　HLP 3P2 1可能为LAK细胞一新的抗菌效应分子。     

Transplantation of human fetal pancreas: fresh vs. cultured fetal islets or ICCS

The paucity of human adult islets available for transplantation in IDDM makes the use of human fetal pancreas a potential alternative. Fetal pancreatic endocrine cells grow and differentiate over time when fresh explants or cultured islet-like cell clusters (ICCs) are transplanted under the kidney capsule in athymic nude mice. We have recently developed a procedure to isolate fetal islets, which differ from ICCs in their β-cell content. This study was undertaken to compare the maturation and growth of grafts from purified fetal islets, containing mostly β-cells, to grafts of mostly undifferentiated endocrine cell precursors, cultured as ICCs, and fresh, uncultured tissue. Total insulin content was highest in the fresh tissue pre-transplant while insulin levels fell precipitously during culture as either fetal islets or ICCs. Although 500 fetal islets contained more insulin than 500 ICCS before transplantation, the insulin content of the resulting grafts was the same 3 months post-transplantation. The degree of stimulation following glucose challenge was comparable, as was the histological appearance. However 70 mg of fresh tissue was needed to generate the fetal islets while only 30 mg was needed for the ICCs. Grafts of 30 mg fresh tissue also had similar total insulin contents and stimulation following glucose challenge, but, when normalized to DNA there was a significantly higher concentration of insulin in the grafts from ICCs or fetal islets. Moreover there were distinct morphological differences, with fibrous and ductal elements prominent in the grafts from fresh tissue, which were also much larger and more diffuse, with cystic elements evident macroscopically. Quantitative immunohistochemical analysis showed that grafts from cultured tissue were 48.3±5% positive for immunoreactive insulin compared with grafts from fresh tissue which were only 13.3±1.4% positive for insulin. In conclusion cultured ICCs, a heterogeneous mixture of hormone-containing and undifferentiated endocrine cells, are a preferable source for transplantation than either purified fetal islets or uncultured tissue.     

人膀胱癌上皮细胞株T24分泌LAK细胞诱导因子（LAK—IF）

从人膀胱上皮癌细胞株T24的亚克隆株T24-8的无血清培养上清液经55%硫酸铵沉淀,再对无离子水透析,得到了有诱导LAK 细胞活性的粗提品。有此作用的因子称为LAK 细胞诱导因子(LAK-IF)。它的分子量为67KD,等电点为pH3.1～3.5。抗人IL-6单抗不能阻断LAK-IF 的作用。LAK-IF 粗提品经Mono-Q 柱快速蛋白液相层析见3个峰。第2峰蛋白质有LAK-IF 活性。     

Enhancement of terminal B lymphocyte differentiation in vitro by fibroblast-like stromal cells from human spleen

Stromal elements are major components of lymphoid tissues contributing to both tissue architecture and function. In this study we report on the phenotype and function of fibroblast-like stromal cells obtained from human spleen. These cells express high levels of CD44 and ICAM-1 and moderate levels of VLA-4, VCAM, CD40 and CD21. They fail to express endothelial, epithelial, lymphocyte and monocyte/macrophage markers. We show that these cells interact with B cell blasts induced in vitro by anti-CD40 and anti-μ stimulation. As a result of these interactions both IL-6 and IgG secretion into culture medium is increased. The enhanced secretion of IgG is partly inhibited by abolishing B cell blast-stromal cell contact or by anti-IL-6, anti-VCAM or anti-CD49d antibodies. Our studies also suggest that the ability of stromal cells to promote B cell survival is most likely the underlying mechanism of the enhanced immunoglobulin secretion. Comparison of stromal cells from different lymphoid and non-lymphoid organs revealed that bone marrow- and spleen-derived stromal cells are the most effective in promoting B cell blast differentiation.     

Application potential of human fetal stem/progenitor cells in cell therapy

We analyzed the possibility of using fetal stem and progenitor cells for the treatment of various pathologies. A comparative characteristic of stem cells from fetuses and adult donors is presented and advantages of the use of fetal biometerial for biotransplantation are described. The main tissue sources of fetal stem cells are characterized and experimental and clinical data on the use of fetal cells for cytotherapy are presented. __________ Translated from Kletochnye Tehnologii v Biologii i Meditsine, No. 1, pp. 5–13, January, 2007     

脾LAK细胞培养上清中BLT酯酶活性的动态观察

脾ＬＡＫ细胞培养上清有前后两个杀伤活性高峰，以第１１ｄ为界，第一个活性高峰与上清中ＢＬＴ酯酶活性呈正相关。含血清和无血清的培养体系产生ＢＬＴ酯酶的机理可能不同。在有高活性ｒＩＬ－２时，ＢＬＴ酯酶的分泌与无血清培养液中有无２－ＭＥ相关。