树突状细胞对LAK细胞抗人鼻咽癌细胞的促进作用

本研究对人血树突状细胞(Dendritic cell,DC)联合LAK(Lymphokine Activated Killer cell,LAK)细胞和IL-2对人鼻咽癌细胞株Hep-2的抗肿瘤活性进行了体外观察。实验分为LAK组、LAK+DC组和LAK+DC+IL-2组;效靶比例分别采用10:1和20:1二种。37℃、5％CO_2、饱湿条件下培养48h后,用中性红摄入比色法检测细胞毒活性。结果:各组的细胞毒活性依次为LAK+DC+IL-2组>LAK+DC组>LAK组(P<0.001),随着效靶比例升高,各组细胞毒活性增强(P<0.01).表明DC和IL-2有协同作用,能增强LAK细胞对Hep-2的细胞毒活性。     

人LAK细胞的体外抗肿瘤作用

用重组IL-2(rIL-2)以及部分纯化的IL-2(PPIL-2)体外激活人外周血单个核细胞.(PBM),使之形成LAK细胞,然后借助于~(51)Cr释放实验,研究了正常人和肿瘤病人的LAk细胞对传代的肿瘤细胞系和新鲜实体瘤细胞的杀伤能力。实验结果表明:1.二种来源的LAK细胞均能明显杀伤传代的肿瘤细胞系,包括NK敏感的K562细胞和NK抵抗的Daudi细胞。2.采用数种新鲜实体瘤细胞作靶,二种来源的LAg细胞同样具有明显的广谱杀伤力,这证实该群杀伤细胞确系LAK细胞。3.不同来源的实体瘤细胞对LAK细胞的杀伤敏感性不同,表现为杀伤程度上的差异,这似乎提示某些肿瘤对LAK细胞杀伤存在抗性。     

IL-2诱导LAK细胞活性但无HNK1阳性细胞数增加

目前多数学者认为,LAK细胞本质上是NK细胞,在IL-2诱导下其细胞毒活性增强,为LAK现象。在LAK细胞研究中,关于IL-2诱导下LAK活性增高与细胞增殖之间的关系,文献报道互有矛盾。鉴于HNK1抗原选择性地表达于NK和K细胞,LAK细胞HNK1亦为刚性,本实验从IL-2诱导前后HNK1阳性率的变化及LAK功能的流式细胞仪分析,探讨了LAK细胞活性与细胞增殖的关系。结果表明,在本实验条件下,IL-2可诱导出LAK活性,但HNK1阳性细胞数不增加,提示LAK细胞活性可能与细胞增殖无关。实验中还观察到去单核细胞使IL-2诱导的LAK活性下降,提示单核细胞在IL-2诱导下也有LAK活性。     

精脒和腐胺对人胎儿脾LAK活性的影响

本研究表明,精脒、腐胺本身均不能使人胎脾单个核细胞(FSMC)转化为LAK细胞,但与5u/ml重组白细胞介素2(rIL-2)伍用,可以增强人胎脾LAK细胞活性;精脒,腐胺不能直接刺激细胞增殖,但能协同亚适剂量Con A诱导淋巴细胞增殖;精脒或腐胺本身可刺激FSMC的IL-2受体(IL-2R)的表达,并能协同rIL-2进一步提高人胎脾LAK细胞IL-2R的表达;精脒、腐胺均可使FSMC及胎脾LAK细胞内鸟氨酸脱羧酶(ODC)活性增强,而以腐胺的作用尤为显著。     

胰岛素对LAK细胞的增殖和杀伤活性的影响

LAK细胞调节因素的研究对揭示LAK细胞的内在规律以及免疫过继疗法的开展与完善均具有重要意义。实验发现胰岛素能促进LAK细胞的增殖,提高细胞的产率,但是胰岛素不能替代IL-2使经短期IL-2刺激的淋巴细胞继续增殖。胰岛素还能促进LAK细胞表面IL-2R的表达。这可能是胰岛素促进LAK细胞增殖的重要机制。     

人胎脾脏和胸腺细胞的LAK活性与增殖特性

本文报道用细胞毒及~3H-TdR掺入等方法,研究人重组IL-2对人胚胎中期脾脏和胸腺细胞LAK活性的诱导及细胞增殖反应影响的一般规律。在低剂量IL-2的作用下,人胎脾脏和胸腺细胞出现广谱杀瘤活性,但二者的活性表达有所不同;脾脏细胞出现很强的增殖反应,胸腺细胞没有出现。实验结果提示,胎儿脾脏和胸腺细胞对重组IL-2的反应性存在差异。     

基因重组肿瘤坏死因子对LAK细胞体内外抗肿瘤作用的增强效果

TNF单独不能诱导LAK活性,也不能进一步提高最适剂量IL-2(1000U/ml)诱导的LAK活性,但能显著增强亚适剂量IL-2(10～100U/ml)诱导的LAK活性。抗TNF单抗和抗IL-2受体β链(p75)单抗显著抑制TNF/IL-2对LAK活性的协同诱导作用。实验证明,TNF能显著增强IL-2/LAK细胞的体内抗肿瘤效果,对于腹水型肝癌小鼠,以联合腹腔内注射TNF/IL-2/LAK细胞的体内抗肿瘤效果最佳。     

IL-2和IL-15协同调节T细胞的增殖和LAK细胞的杀伤活性

目的 探讨IL-2和IL-15在免疫调控和应答中的协同作用。方法 IL-2、IL-15和IL-2 IL-l5组刺激CTLL细胞的增殖，^3H—TdR掺入法测cpm值；IL-2、IL-15和IL-2 IL-l5组诱导PBMC中NK和LAK细胞的发育，4h^51Cr释放实验检测对K562和LiBr细胞的杀伤活性。结果 IL-2和IL-15都能诱导CTLL认细胞的增殖和NK、LAK细胞的杀伤活性，IL-2和IL-15可明显协同上调CTLL认的增殖和LAK细胞的杀伤活性。结论 IL-2和IL-15在免疫调控和应答中有一定的协同作用，为细胞因子联合应用于临床治疗肿瘤等疾病提供了实验依据。     

癌症的淋巴因子基因疗法

一、淋巴因子免疫疗法最近,重组淋巴因子的克隆、表达和大规模制备大大促进了辅助性T 细胞依赖性效应细胞的开发应用。尤其是重组IL-2已在体外用于诱导LAK 细胞,且能使转移性恶性黑色素瘤、肾细胞癌等某些肿瘤得以缓解。IL-2扩增后的TILS(肿瘤侵润性淋巴细胞),包括LAK 细胞前体、细胞毒性T 细胞和辅助性T 细胞,对同系肿瘤明显具有比LKA 细胞更强的细胞毒性,从而具有更大的疗效潜力。很多淋巴因子,包括IL-1、LL-4、IL-5、TNF、GM-CSF 和IFN—β在体外诱导LAK 细胞中与IL-2有协同作用,从而为淋巴因子联合应用的体内试验开     

白细胞介素2治疗前后甲襞微循环观察

<正> 自细胞介素2(Inter Leu kin2 IL-2)又称T细胞生长因子(TCGF)。是T淋巴细胞受抗原刺激后所分泌的具有免疫调节作用的淋巴因子。IL-2诱导自然杀伤细胞(NK)、细胞毒性T细胞(CTL)、淋巴因子活化的杀伤细胞(LAK)的活化和增殖,从而杀伤肿瘤细胞,的作用近年来日益受到人们的重视。联合应用大剂量IL-2和LAK细胞即IL-2/LAK疗法,治疗多种肿瘤患者已取得了显著的效果。但     

酵母菌重组白细胞介素2生物学活性鉴定

利用IL-2依赖细胞系CTLL检测酵母菌系统表达的人rIL-2生物活性。结果证实,酵母菌 rIL-2 能明显刺激 CTLL细胞的生长。用进口的 rIL-2作为标准,测定所表达的 rIL-2活性单位为 33u/ml。单克隆抗体中和实验证实,酵母菌 rIL-2 能特异地和天然 IL-2单抗结合,抑制其本身诱导的 CTLL细胞增殖。这种抑制是由 IL-2单抗参与的,因为鼠正常血清没有这种抑制作用。此外,经蛋白A 柱层析纯化后的单抗在 0.27μg/孔时就能表现明显地抑制,提示该抗原抗体反应的特异性和专一性。     

转移因子和免疫核糖核酸对LAK细胞抗肿瘤活性的影响

本研究用重组白细胞介素２（ｒＩＬ－２）联合转移因子（ＴＦ）或抗肿瘤免疫核糖核酸（ｉＲＮＡ）作用于外周血单个核细胞（ＰＢＭＣ），测定其对Ｋ５６２，Ｒａｊｉ及Ｈ７４０４细胞的杀伤活性。结果发现，高浓度ＴＦ（０．２５ｕ／ｍｌ）可抑制ＬＡＫ活性，ＴＦ进一步稀释（０．１２５ｕ／ｍｌ）可使其抑制作用消失。ＴＦ和抗肿瘤ｉＲＮＡ不能进一步提高最适剂量ｒＩＬ－２（５００ｕ／ｍｌ）诱导的ＬＡＫ活性，但能显著增强亚适剂量ｒＩＬ－２（２００ｕ／ｍｌ）诱导的ＬＡＫ活性，诱导ＬＡＫ活性增高的ＴＦ最适剂量为０．０３１ｕ／ｍｌ。ＴＦ或抗肿瘤ｉＲＮＡ单独不能诱导出ＬＡＫ活性，而当两者联合应用可诱导ＰＢＭＣ产生ＬＡＫ活性。本文为ＴＦ及抗肿瘤ｉＲＮＡ协同ＬＡＫ细胞疗法在临床应用提供实验基础。     

Decreased cell replication and polyamine content in insulin-producing cells after exposure to human interleukin 1 beta

Interleukin 1 (IL-1) has been suggested to cause the islet B cell destruction occurring during the development of insulin-dependent diabetes mellitus. One mechanism by which B cell loss can be compensated for is via de novo formation of new cells through replication. In the present study the replicatory activity of cells in isolated rat pancreatic islets and in the insulin-producing cell line RINm5F has been assessed by [3H]thymidine incorporation methods after exposure to 1-25 U/ml of human recombinant IL-1 beta (rIL-1 beta). In the rat islets [3H]thymidine incorporation was decreased by 20% 5 h after exposure to 25 U/ml rIL-1 beta. A similar inhibition was also observed in islets exposed to 2.5 and 12.5 U/ml rIL-1 beta. In the RINm5F cells there was a dose-dependent inhibition of the cell replication to approximately 50% of the controls in cells exposed to 25 U/ml rIL-1 beta for 48 h. This was also accompanied by an increased cell death, as measured by trypan blue inclusion (controls 13% and rIL-1 beta treated cells 25%). The insulin content of the RINm5F cells was reduced by about 40% after a 48-h exposure to 25 U/ml rIL-1 beta. When the exposure of the RINm5F cells to rIL-1 beta was decreased to 24 h there was no increased cell death, but a reduced replicatory activity was still observed. rIL-1 beta decreased the cellular content of the polyamines spermidine and spermine.(ABSTRACT TRUNCATED AT 250 WORDS)     

Differentiation of Macrophage Precursors to Cells with LAK Activity under the Influence of CSF-1 and High Dose IL-2

Mouse macrophage precursor cells with natural killer (NK) like activity, derived from in vitro culture of light-fraction-bone marrow cells in the presence of colony-stimulating factor 1 (CSF-1) and low dose IL-2, were incubated with high dose (1000 U/ml) IL-2. After 3 days of incubation, cells had developed from NK like (killing Yac-1 but not P815) into LAK-like (killing both YAC-1 and P815) effector cells. Morphological studies revealed that LAK activity occurred at the time when macrophage precursors with NK like activity containing few cytoplasmic granules had further differentiated into cells with abundant azurophilic granules in their cytoplasma. Proliferation of macrophage-precursor derived NK/LAK-like cells was dependent on the presence of colony-stimulating factor, generation of cytoplasmic granules was induced by IL-2 in a dose dependent way. Flow cytometric analysis showed that macrophage precursor-derived LAK effectors were positive for NK 1.1 but almost negative for F4/80. When the same starting cell population was cultured in the presence of 200 U/ml Interferon gamma (IFN gamma), proliferation was completely stopped and within 3-4 days all cells differentiated into mature macrophages expressing F4/80. In context with our previous data, we describe here a continuum of development from agranular macrophage precursors to granular cells with NK-like activity and further to cells with LAK activity under the influence of CSF-1 as growth factor and IL-2 as granule- and cytotoxicity-inducing factor.     

国产rIL-2诱导LAK作实验性抗肿瘤过继免疫治疗

肿瘤细胞体外杀伤实验表明,应用1000u/ml国产重组白细胞介素-2(rL-2)体外诱导6天产生的LAK细胞可获最佳杀伤效果,应用这一诱导条件并参照临床过继性免疫治疗的“3×5”方案,合并IL-2对Balb/c小鼠体内鼠肝癌及裸鼠体内人胃癌移植肿瘤作过继性免疫治疗,生长期肿瘤的总抑瘤率分别达到68.7%和52.4%,表明该剂量的国产rIL-2在实验性抗肿瘤过继免疫治疗中是有效的和安全的。     

The biological effects of rainbow trout (Oncorhynchus mykiss) recombinant interleukin-8

In this report, recombinant interleukin-8 (rIL-8) was produced and its activity tested for the first time in fish. The rainbow trout rIL-8 was produced in Escherichia coli and purified using a 6xHis tag at the N-terminus. The rIL-8 induced a dose-dependent migration of head kidney leukocytes at concentrations from 0.1 to 10 ng/ml, with a peak response at 1 ng/ml. Trout rIL-8 also had a significant effect on superoxide production by head kidney cells, with maximal activity at 0.1 and 1 ng/ml. When injected intraperitoneally into trout, rIL-8 had a clear effect on total leukocyte number in the peritoneal cavity, with increasing doses (up to 5 microg) eliciting more cells. Of three leukocyte types distinguished, neutrophils were the dominant cell type, especially at higher rIL-8 concentrations. In contrast, the proportion of macrophages and lymphocytes decreased with rIL-8 administration, suggesting that they were not attracted at the same rate as neutrophils.     

生物素-链霉亲和素系统在检测人PBMC IL-2R_α中的应用

本文报道用自制生物素-链霉亲和素系统配以抗IL-2R_aMcAb,建立检测人PBMCIL-2R_a的方法,并对本法中的关键条件进行探索,确定生物素化二抗和链霉亲和素的最适浓度以及多种诱导物的最适诱导时间和浓度。用本法对比检测了健康献血者和肿瘤患者PBMC IL-2R_a的表达水平,表明未经诱导的肿瘤患者PBMC IL-2R_a百分率显著高于正常人(P<0.01)。     

Population dynamics of natural killer cells in the spleen and bone marrow of normal and leukemic mice during in vivo exposure to interleukin-2.

By quantitative and functional methods, changes were assessed in NK(ASGM-1+) cell numbers and NK cell-mediated lytic function of the spleen and bone marrow of mice bearing a tumor of hemopoietic origin (FLV-induced erythroleukemia) for 9 days +/- simultaneous administration of indomethacin (10 micrograms/ml drinking water) +/- rIL-2 (3x/day, 12 x 10(3) Units/injection) during the last 4 days of tumor-bearing. Recombinant IL-2 alone during the last 4 days of tumor-bearing increased both the NK(ASGM-1+) cell numbers (p less than 0.001) and the functional activity (24-fold) of the spleen. In the bone marrow, however, no change in the numbers of NK(ASGM-1+) cells was observed relative to untreated tumor-bearing mice, but the NK cell-mediated lytic activity of that organ was augmented 30-fold. The continuous presence of indomethacin from the onset of tumor-bearing prior to rIL-2 treatment during the last 4 days of tumor-bearing, further boosted both the already high, rIL-2 driven numbers of NK(ASGM-1+) cells in the spleen (p less than 0.01), as well as splenic NK cell lytic function (2-fold). In the bone marrow, continuous presence of indomethacin prior to and during the terminal 4 days of co-administration with rIL-2 increased 3-fold the numbers of NK(ASGM-1+) cells relative to that of the bone marrow of tumor-bearing mice given rIL-2 alone, and resulted in lytic activity of that organ which was 140% of that of the rIL-2 treated, tumor-bearing mice. The results indicate that under the combined influence of indomethacin and rIL-2, the production of NK(ASGM-1+) cells was augmented in the bone marrow of tumor-bearing mice, export of immature NK(ASGM-1+) cells from the bone marrow was increased, and import of immature NK(ASGM-1+) cells by the spleen was increased. The increased NK(ASGM-1+) cell numbers in each organ was reflected in increased lytic function.     

Recombinant interleukin 2 inhibits pokeweed mitogen-induced proliferation of human adult peripheral blood and cord blood mononuclear cells

We studied the effects of recombinant interleukin 2 (rIL-2) on pokeweed mitogen (PWM)-induced proliferation of unfractionated human mononuclear cells from the peripheral blood (PBMCs) and cord blood (CBMCs). rIL-2 was found to inhibit PWM-induced proliferation of both PBMCs and CBMCs in a dose-dependent manner. The response was statistically significant at concentration 10 U/ml. The inhibition of adult PBMC proliferation was relatively mild, but in newborns up to 75% inhibition was found. The inhibitory effect of rIL-2 was best detectable at day 4, at the same time as PWM-induced proliferation peaked. However, also at days 8 and 10, the response induced by PWM plus rIL-2 was much lower than that induced by rIL-2 alone. These finding suggest that the inhibition was due to rIL-2-mediated stimulation of the function of PWM-activated suppressor cells. CD8+ T cells were, however, not alone responsible for the rIL-2-induced inhibition of PWM-induced proliferation, since the responses were essentially similar independent of whether CD8+ T cells were present or absent. In addition, the release of interferon-gamma (IFN-gamma) in response to rIL-2 was not a major cause of the inhibitory effects, because rIL-2 inhibited PWM-induced proliferation of CBMCs, which are deficient in producing IFN-gamma. Our results provide novel data to the ongoing discussion of IL-2 as a down-regulator of immune functions.     

Effect of recombinant human interleukin 2 (rIL-2) on normal peripheral blood B cells and B lymphoblastoid cell lines

The role of interleukin 2 (IL-2) for growth and differentiation of normal and malignant B cells still remains controversial. We assessed normal peripheral blood B cells and cell lines derived from patients with B non-Hodgkin's lymphomas (NHLs) with respect to their responsiveness to recombinant human IL-2 (rIL-2). The NHL cell lines used in our experiments expressed the Tac antigen (CD25)--a compound of the IL-2 receptor (IL-2R)--in a percentage ranging from 28 to 57%. As measured in a [3H]thymidine uptake assay, the normal peripheral blood B cells demonstrated a dose-dependent proliferative response to rIL-2, whereas the NHL cells did not show any responsiveness to rIL-2. In a clonogenic culture assay we evaluated the colony formation of the NHL cells and found a decrease of 28 to 41% on average in the presence of rIL-2 (10-50 U/ml). This moderate inhibitory effect on the clonal growth of the NHL cells was not due to a differentiation inducing effect of rIL-2, as studied by measuring the Ig production under increasing doses of rIL-2 (1 to 100 U/ml). Thus, malignant NHL B cells may express the CD25 compound of the IL-2 receptor on their surface, demonstrating a different functional responsiveness to rIL-2 compared to normal peripheral blood B cells.