多种活化因子共刺激对人外周血单个核细胞体外增殖和表型的影响

目的:观察多种活化因子共刺激后对人外周血淋巴细胞增殖和表型的影响。方法:用淋巴细胞分离液分离人外周血单个核细胞(PBMC),根据加入刺激因子(CD3 mAb、CD28 mAb、IFN-γ、IL-1α、IL-2、IL-15和IL-21)种类和方法不同将实验分为7组。用全自动五分类血液分析仪计数不同细胞因子诱导培养的PBMC增殖力、流式细胞术测定诱导后共刺激细胞表面CD3,CD4,CD8,CD28,CD16、CD56+CD16,CD3+CD8+,CD3+CD4+,CD3+CD56+,CD45RO等分子的变化、乳酸脱氢酶释放法测定诱导后的共刺激细胞对SGC-7901、SW-1990和SW-116细胞株的杀伤活性。结果:在PB-MC培养体系中加入不同的刺激因子其细胞增殖能力有明显的差异,以含刺激因子CD3、CD28、IFN-γ、IL-2、IL-1α、IL-15和IL-21组增殖倍数最高,在培养第10 d时该组的增殖倍数为255.3 6.3,明显高于仅含CD3、IFN-γ和IL-2培养体系组(166.6 5.5)(P<0.05)。在PBMC培养体系中加入不同的刺激因子其部分细胞表面标志有所差异,在培养体系中无IL-15时CD16+CD56+(NK细胞)细胞和CD3+CD56+细胞比例明显高于其他组;CD45RO+的记忆性T细胞以延迟3 d添加IL-15和IL-21组升高最明显。经不同活化因子刺激培养10 d的PBMC对SGC-7901、SW-1990和SW-1116细胞杀伤活性有明显差异,以延迟3 d添加IL-15和IL-21组最高(分别为76.2%、60.3%和70.6%),明显高于仅含CD3、IFN-γ和IL-2培养体系的细胞组(分别为54.9%、44.6%和50.4%)(P<0.05)。培养的细胞对胃腺癌细胞SGC-7901杀伤活性最强。结论:不同刺激因子活化的PBMC其增殖能力、表面标记和杀伤活性有明显差异,在培养体系中增加相应的刺激因子对细胞定向培养有一定价值。     

IL-23单独或联合IL-2对人PBMC增殖及抗白血病活性影响的研究

目的比较单独应用IL-23或联合应用IL-23和IL-2对人外周血单个核细胞(PBMC)增殖及抗白血病活性的影响。方法密度梯度离心法分离人PBMC,以不同的细胞因子培养液培养,MTT比色法测定PBMC的增殖情况及PBMC对白血病K562细胞株的杀伤活性,流式细胞术分析诱导前后PBMC的细胞表型变化。结果细胞因子各组培养PBMC 1 d、3 d、5 d后,PBMC的增殖以及对K562细胞的杀伤活性均增强(P<0.05),IL-23与IL-2联合后PBMC的增殖以及杀伤活性进一步增强(P<0.05);IL-23(50 ng/ml)与IL-23+IL-2(50 ng/ml+100 IU/ml)组作用于PBMC 5 d后,检测CD3+细胞为(80.7±4.37)%和(83.2±4.04)%,CD16+CD56+细胞为(8.4±0.28)%和(12.7±0.9)%,CD4+细胞为(45.2±1.4)%和(47.0±1.72)%,CD8+细胞为(34.5±2.53)%和(35.4±2.11)%;与对照组相比,两组对PBMC细胞表型的增加有显著性差异(P<0.05);两组间比较CD16+CD56+细胞的表达上有显著性差异(P<0.05),对CD3+、CD4+、CD8+细胞的表达以及CD4+/CD8+比值的变化无显著性差异(P>0.05)。结论 IL-23对PBMC的增殖,以及PBMC对K562细胞的杀伤均有促进作用,与IL-2联合有协同作用。IL-23单独或联合IL-2诱导后PBMC中CD3、CD16/56、CD4、CD8抗原的表达均增加,二者联合后可以进一步增加CD16/56抗原的表达。     

抗原负载的DC诱导CIK对MDA-MB-231细胞的杀伤活性研究

探讨乳腺癌相关抗原负载后树突状细胞(DC)联合细胞因子诱导的杀伤细胞(CIK)对MDA-MB-231乳腺癌细胞株的杀伤效应。采用密度梯度离心法获取外周血单个核细胞(PBMC),常规法诱导DC、CIK细胞;流式细胞技术分析测定细胞表型;用反复冻融法制备MDA-MB-231细胞的相关抗原,并测定抗原浓度;MTT法分别测定在四个不同效靶比时CIK、DC+CIK和抗原负载的DC+CIK三组效应细胞对MDA-MB-231乳腺癌细胞的杀伤活性。结果:CIK细胞分别与DC和Ag-DC共培养后,可见CD3+CD56+双阳性表达细胞较培养前显著增加(P<0.05);将负载肿瘤相关抗原的DC与CIK共培养后,杀瘤活性明显提高(P<0.05)。提示,抗原负载的DC与CIK细胞共同孵育培养后,在相同效靶比时,对MDA-MB-231乳腺癌细胞杀伤效应与CIK细胞和DC-CIK两组效应细胞相比有显著提高,表明抗原负载后可提高CIK细胞对肿瘤细胞的杀伤活性,为乳腺癌临床生物免疫治疗提供实验室基础。     

CD3AK细胞的诱导及其抗肿瘤作用机制的初步探讨

采用两种状态的IgG2型小鼠抗人CD3单抗,辅以小剂量外源性IL-2刺激诱导CD3AK细胞,测定其对K562,Raji及P815细胞的杀伤作用。结果发现,0.1μg/ml浓度的游离状态抗CD3单抗和10μg/ml浓度的粘附状态抗CD3单抗刺激人PBMC增殖达最强效果,其中粘附状态抗CD3单抗诱导的PBMC增殖活性明显强于激离状态抗CD3单抗;外源性IL-2能协同增强抗CD3单抗诱导PBMC增殖,但当粘附状态抗CD3单抗处于最强激活浓度时,其协同作用并不明显。实验结果显示,CD3AK细胞对三种不同性质的肿瘤细胞均具有较强的杀伤作用,其杀伤活性既有效靶细胞直接接触杀伤,又有杀伤因子介导。     

抗原致敏的DC-CIK对前列腺癌细胞DU145杀伤的作用

目的探讨抗原致敏的DC与CIK细胞共同培养后获得的DC-CIK细胞(Ag-DC-CIK)对前列腺癌细胞株DU145的杀伤作用。方法分离健康志愿者外周血单个核细胞,贴壁细胞以GM-CSF、IL-4及TNF-α诱导并加入DU145全细胞冻融抗原培养抗原致敏的DC,非贴壁细胞以IL-2、IL-1α、IFN-γ和mAb诱导培养CIK细胞,并将两种细胞共同培养6d获得Ag-DC-CIK;每隔2天计数细胞;流式细胞术检测细胞免疫表型;以共培养6d的Ag-DC-CIK作为效应细胞,以相同天数未致敏的DC-CIK和CIK细胞为对照组,DU145细胞作为靶细胞,采用MTT法分析各组杀瘤率。结果外周血单核细胞成功诱导DC,Ag-DC-CIK细胞培养6d的扩增倍数为CIK的1.69-2.56倍(P<0.05)。CIK组、DC-CIK组及Ag-DC-CIK组中CD3+CD56+细胞所占百分比分别为(19.15±1.55)%,(28.43±1.51)%和(39.12±2.29)%,依次升高(P<0.01),对前列腺癌DU-145细胞24h杀瘤率分别为(26.39±4.47)%,(46.82±5.68)%,(62.80±2.01)%,依次升高(P<0.01)。结论 Ag-DC-CIK细胞增殖活性强,对前列腺癌DU-145细胞有明显杀伤作用,强于未致敏DC-CIK细胞和CIK细胞。     

IL-33增强体外培养的外周血单个核细胞细胞因子的分泌以及杀伤功能

目的探讨白细胞介素33(IL-33)是否能在体外增强外周血单个核细胞(PBMC)分泌细胞因子和杀伤功能。方法常规分离培养健康人PBMC,分别经CD3单克隆抗体(m Ab)/CD28mAb/IL-2、CD3mAb/CD28mAb/IL-2/IL-12、CD3mAb/CD28m Ab/IL-2/IL-12/IL-33三种组合刺激培养72 h,收集细胞。经实时定量PCR(qRT-PCR)检测各组细胞γ干扰素(IFN-γ)、颗粒酶B(granzyme B)mRNA的水平;CCK-8法检测各组细胞对A549人肺腺癌细胞的杀伤活性;采用流式细胞术分析各组细胞IFN-γ、程序性死亡蛋白1(PD-1)的水平。结果各组因子组合刺激的PBMC,形态无明显差异;qRT-PCR检测发现,IL-33刺激组的PBMC中IFN-γ、granzyme B mRNA的水平升高;IL-33刺激组的PBMC对A549的杀伤能力更强;IL-33刺激组的PBMC中CD3+CD8+细胞亚群IFN-γ的水平升高、PD-1的水平降低。结论 IL-33能够在体外增强PBMC的效应功能。     

脐血CIK细胞诱导K562细胞的凋亡作用

探讨脐血CIK细胞对K562细胞的杀伤活性及诱导凋亡作用。通过第1天在脐血淋巴细胞中加入IFN-r，第2天再加入IL-2和CD3单抗进行细胞培养获得多种细胞因子诱导的杀伤细胞(Cytokines induced killer cells)即脐血CIK细胞。用MTT法测定其抗肿瘤活性，原位末端标记法进行凋亡分析，台盼蓝拒染法计数细胞。脐血CIK细胞及培养上清抗K562活性明显优于CD3AK细胞，短期培养后其增殖活性低于CD3AK细胞；但是加入G-CSF可以提高CIK细胞的增殖活性，且CIK细胞的抗K562活性仍高于CD3AK细胞。CIK细胞诱导K562细胞凋亡率大于CD3AK细胞，G-CSF能部分抑制CIK细胞诱导的K562细胞的凋亡。脐血CIK细胞是一种有效的杀伤活化细胞，脐血在多种细胞因子适当组合的诱导下可提高杀瘤活性。     

IL-21增强γδT细胞体外抗肿瘤活性的实验研究

探讨IL-21对人外周血γδT细胞抗肿瘤活性的影响。分离健康人外周血单个核细胞(PBMC),分别在有或无IL-21参与的条件下,加入到含异戊烯焦磷酸(isopentenyl pyrophosphate,IPP)、IL-2或IL-15的RPMI 1640完全培养基中诱导培养γδT细胞。在培养的第10天用台盼蓝染色计数细胞;采用流式细胞术检测各组γδT细胞的纯度及γδT细胞穿孔素、颗粒酶B和CD107a的表达;用CCK-8试剂盒检测γδT细胞对K562细胞的体外杀伤效应。结果显示,不同细胞因子组诱导10d后γδT细胞纯度均达到70%以上,IL-2+IL-21、IL-15+IL-21和IL-2+IL-15+IL-21组与IL-2、IL-15组相比,细胞纯度和扩增倍数没有显著性变化;IL-2+IL-21、IL-15+IL-21和IL-2+IL-15+IL-21组γδT细胞穿孔素、颗粒酶B和CD107a的表达及对K562细胞的杀伤活性均显著高于IL-2、IL-15单独组。以上结果表明,IL-21可通过上调γδT细胞穿孔素和颗粒酶B的表达增强其抗肿瘤活性。     

DC-CIK细胞体外抗淋巴瘤细胞的免疫效应研究

目的:探讨共培养树突状细胞(Dendritic cell,DC)和细胞因子诱导的杀伤细胞(Cytokine-induced killer cell,CIK),即产生的DC-CIK细胞抗淋巴瘤免疫效应的机制。方法:分离健康人外周血单个核细胞,分别于体外诱导DC和CIK,然后共培养成DC-CIK细胞。分四组:DC组、DC-T组、CIK组和DC-CIK组。对各组分别进行免疫机制研究:测定细胞因子IL-12、IL-17和IFN-γ的分泌量、检测细胞免疫表型和分析对人淋巴瘤细胞株杀伤活性。结果:在DC-CIK组,细胞因子IL-12、IL-17和IFN-γ的分泌量均明显高于其它三组(P0.05);杀伤活性亦较其它组增强(P0.05)。CD8+、CD3+/CD56+细胞在DC-CIK组亦明显增加(P0.05)。结论:DC-CIK细胞抗淋巴瘤免疫效应与分泌大量的细胞因子和产生细胞毒性细胞有关。     

IL-24基因修饰的树突状细胞促进CIK细胞对肺癌细胞的杀伤作用

目的 研究IL-24基因修饰的树突状细胞(DC)与同源细胞因子诱导的杀伤细胞(cyto-kine-induced killer,CIK)共培养后对A549肺癌细胞的杀伤作用及其机制.方法 从健康人外周血单个核细胞中常规诱导DC、CIK细胞,同时抽提重组腺病毒质粒pAdEasy-1-pTrack-CMV-IL-24,Pac I酶切线性化后脂质体转染QBl-293A细胞,收获Ad-IL-24重组病毒.将IL-24基因通过重组病毒导入已负载肿瘤抗原的DC,获得细胞称为DC-IL-24.RT-PCR和ELISA法检测DC中IL-24基因的表达,FCM和ELISA法检测DC表型及分泌细胞因子能力的变化,将DC和CIK细胞混合培养,溶血试验检测CIK细胞产生穿孔素的能力,FCM法检测共培养的DC-CIK细胞对A549肺癌细胞细胞毒活性的变化.结果 获得了高滴度的重组病毒Ad-IL-24并成功将IL-24基因导入DC,在倒置荧光显微镜下可观察到荧光,IL-24可上调Dc表而CD80、CD83、HLA-DR、CD40、CXCR4分子的表达,转染后Dc分泌IL-12、TNF-α和IL-24的能力显著增强,DC-IL-24更能促进CIK细胞产生穿孔素,与同源CIK细胞共培养后对A549肺癌细胞的细胞毒活件明显增强.结论 IL-24基因修饰的DC能增强自体CIK细胞产生特异性抗肿瘤免疫,其机制与IL-24能促进DC表型成熟,分泌Th1型细胞因子,维持DC活化状态,进而促进CIK细胞活化密切相关.     

Interleukin-15 enhances cytokine induced killer (CIK) cytotoxic potential against epithelial cancer cell lines via an innate pathway

CIK cells are a subset of effector lymphocytes endowed with a non-MHC restricted anti-tumor activity making them an appealing and promising cell population for adoptive immunotherapy. CIK are usually generated ex-vivo by initial priming with Interferon-γ (IFN-γ) and monoclonal antibody against CD3 (anti-CD3), followed by culture in medium containing Interleukin-2 (IL-2). Interleukin-15 (IL-15) shares with IL-2 similar biological functions and recently it has been reported to induce CIK with increased anti-leukemic potential.The aim of the study was to compare the killing efficacy of CIK generated by IL-2 alone or IL-2 and IL-15 toward tumor targets of different origins, leukemic cells and malignant cells from epithelial solid tumors. CIK bulk cultures were examined for cell proliferation, surface phenotype and cytotoxic potential against tumor cell lines K562, HL60, HeLa and MCF-7. The results showed that IL-15 is able to induce a selective expansion of CIK cells, but it is less effective in sustaining CIK cell proliferation compared to IL-2. Conversely, our data confirm and reinforce the feature of IL-15 to induce CIK cells with a potent cytotoxic activity mostly against tumor cells from epithelial solid malignancies via NKG2D-mediated mechanism.     

PD-L1-GPI分子膜表面修饰胰岛细胞抑制反应性T细胞的攻击

目的： 构建PD-L1表达腺病毒载体,研究其对反应性T细胞的免疫抑制效应,以探讨预防1型糖尿病患者的胰岛β细胞的免疫损伤机制。方法: 用含目的基因的腺病毒载体pAd5/ PD-L1-GPI感染小鼠胰岛瘤细胞株NIT细胞, 提取膜蛋白经Western blotting证实靶细胞高效表达目的蛋白PD-L1-GPI。取小鼠外周血非贴壁的淋巴细胞, 与靶细胞混合淋巴细胞培养,进行淋巴细胞增殖和反应性T细胞功能检测,并检测细胞培养液中IL-2、TNF-α和IFN-γ水平。 结果: 与对照组NIT细胞比较,用PD-L1-GPI修饰的NIT细胞能够更有效地抑制已致敏的淋巴细胞增殖,且下调反应性T细胞分泌的IL-2、IFN-γ和TNF-α等细胞因子水平(P<0.05)。 结论: PD-L1基因在胰岛细胞中表达可有效地抑制已活化的反应性T细胞,阻断自身免疫性T细胞的胞毒作用,诱导免疫耐受,将成为治疗糖尿病的一个有价值的方向。     

细胞因子诱导的杀伤细胞对卵巢癌耐药细胞SKOV3/CDDP的作用机制

目的：探讨卵巢癌耐药细胞在癌变过程中免疫逃逸的分子机制以及细胞因子诱导的杀伤细胞（CIK）对其耐药性及免疫原性的影响。方法：采用电镜、流式细胞术（FCM）、MTT等检测方法，观察CIK细胞作用人卵巢癌耐药细胞系SKOV3／CDDP细胞超微结构、细胞周期、凋亡率、耐药相关基因MDR1和Topo—Ⅱβ以及hB7-1、hB7-2、MHCⅠb、HLA—DR抗原表达的变化；应用放射免疫（RIA）ELISA等方法检测CIK细胞作用于和荷人卵巢癌SKOV3／CDDP耐药细胞SCID小鼠腹腔移植模型后血清IL-2、TNF—α、IFN-γ、GM—CSF等细胞因子含量的改变。结果：电镜显示：CIK组可见较多的凋亡细胞，表现为细胞体积萎缩，膜发泡，细胞密度增加，线粒体浓缩染色质固缩于核膜周边；与生理盐水组相比，CIK组的G0／G1期细胞比例下降，S＋G2／M期细胞比例升高，细胞周期阻滞于S＋G2／M期，有统计学意义（P〈0．05）；在抑制细胞增殖的同时，尚诱导细胞凋亡，其细胞凋亡率为9．07％，二者比较，有统计学意义（P〈0．05）；与NS组相比，CIK组细胞MDR-1和Topo—Ⅱβ表达明显下降（P〈0．05），提示CIK细胞可逆转人卵巢癌耐药细胞SKOV3／CDDP细胞耐药相关基因MDRI和Topo—Ⅱβ的表达；与NS组相比，CIK组细胞MHCⅠb、HLA—DR、hB7—1和hB7-2抗原的表达均明显升高（P〈0．01）。提示CIK细胞可提高人卵巢癌耐药细胞株SKOV3／CDDP的免疫原性；与NS组相比，CIK组小鼠血清中IL-2、TNF-α、INF-γ、GM—CSF含量均明显增加（P〈0．01）。结论：CIK细胞在将卵巢癌耐药细胞的细胞周期阻滞于S＋G2／M期的同时，尚能诱导其凋亡，降低耐药基因的表达，提高耐药细胞的免疫原性，分泌IL-2、TNF-α、INF-γ、GM-CSF等具有抗肿瘤活性的细胞因子发挥抗肿瘤作用。     

Expansion of cytotoxic CD3+ CD56+ cells from peripheral blood progenitor cells of patients undergoing autologous hematopoietic cell transplantation.

Immunotherapy may potentially improve the outcome of autologous hematopoietic cell transplantation (HCT). Poor effector cell proliferation and marginal antitumor activity limit attempts to use immunotherapy. We have characterized the ex vivo expansion, up to 1000-fold, of CD3+ CD56+ lymphocytes from the peripheral blood lymphocytes (PBL) of healthy donors. Expanded cells termed cytokine-induced killer (CIK) cells induce non-major histocompatibility complex-restricted lysis of tumor cells and demonstrate cytolytic activity superior to lymphokine-activated killer cells without the requirement of interleukin (IL)-2 treatment in vivo. To determine whether cytolytic cells could be expanded from patient material, we evaluated samples of peripheral blood progenitor cells (PBPCs) from 25 patients undergoing autologous HCT. The PBPCs were expanded by priming with interferon-gamma followed by anti-CD3 monoclonal antibody and IL-2 the next day. Fluorescence-activated cell sorting analysis was performed on days 0, 15, 21, and 28 of cell culture. The median T-cell content rose from 15.3% (range, 1.1% to 89.7%) on day 0 to 97.2% (range, 83.6% to 99.5%) by day 15. By day 21, T cells expanded 21.8-fold (range, 1.7- to 420.0-fold) and CD3+ CD56+ cells expanded 44.8-fold (range, 5.1- to 747.0-fold). CIK cells were used as effector cells against B-cell lymphoma targets (OCI-Ly8) with a median of 24% (range, 3% to 67%) and 42% (range, 6% to 96%) specific lysis of target cells on days 21 and 28, respectively. CIK cells derived from PBL of 2 additional patients with acute myelogenous leukemia demonstrated 39% and 78% specific lysis of OCI-Ly8 and 26% and 58% specific lysis of autologous leukemic blasts at an effector:target ratio of 40:1. CIK cells may be expanded from granulocyte colony-stimulating factor-mobilized PBPCs of patients undergoing autologous HCT. CIK cells may provide a potent tool for use in posttransplantation adoptive immunotherapy.     

NK 细胞的高效扩增及其对体外肿瘤模型的杀伤效果研究

目的:高效扩增NK 细胞,并确定其对不同肿瘤的杀伤作用,为临床应用提供参考。方法:从成人外周血中分离PBMCs 并利用表面表达4-1BBL、IL-15 和IL-21 的K562 滋养细胞对其进行共刺激培养,15 d 后计数并检测CD3- CD56+细胞纯度;利用间接免疫荧光及Real-time PCR 方法检测NK 细胞Granzyme B 及Perforin 表达变化的同时检测其对肺癌、胃癌、肝癌、卵巢癌、胰腺癌、宫颈癌的杀伤作用,确定其对不同肿瘤的杀伤效果。结果:扩增15 d 后,细胞数量高于110 ‘,CD3-CD56+比例达95%以上;培养后的NK 细胞高表达Granzyme B、Perforin,且对A549 的杀伤效果(E:T=10 :1)约为90%,同时对其他肿瘤细胞(E:T=10:1)的杀伤效果由强到弱的顺序为:胃癌、胰腺癌、宫颈癌、卵巢癌、肾癌、肝癌(P<0.05);NK 细胞对宫颈癌、肝癌、胰腺癌的杀伤效果呈现一定的时间依赖性。结论:此方法可以扩增出高数量、高纯度的NK 细胞,同时该NK 细胞具有高效杀伤多种肿瘤细胞的能力。     

细胞因子诱导的杀伤细胞的生物学特性研究

目的:探讨细胞因子诱导的杀伤细胞(CIK)的生物学特性。方法:健康人非贴壁单个核细胞用含IFN-γ、IL-1β、IL-2、CD3单抗诱导CIK细胞。以LAK细胞作为对照。流式细胞仪和免疫细胞化学染色检测细胞表型特征;乳酸脱氢酶释放法分析细胞毒活性。结果:诱导2周后,CIK细胞的增殖率达到高峰,CD3⁺细胞占95％以上;第3周细胞生长进入平台期。诱导15d时,CD3⁺CD56⁺NKT亚群占16.5％,比例在2-4周区间内无明显变化。LAK细胞增殖缓慢,显著低于同期CIK细胞增殖率(P＜0.01)。不同效:靶比例CIK细胞对肝癌细胞BeL-7402的特异性溶解率显著高于LAK细胞(P＜0.01)。免疫细胞化学染色结果显示,CIK细胞高度表达HLA-DR和CD54抗原,NKT细胞体积较CD3⁺CD56^-细胞略大,细胞表面有大量伪足。结论:CIK细胞具有高度增殖能力,其体外杀瘤活性明显优于LAK细胞。诱导14-21d,CIK细胞增殖率和CD3⁺CD56⁺阳性细胞率均达到高峰,此时期的CIK细胞适合临床应用。     

Effect of CD80 and CD86 on T Cell Cytokine Production

Conjugation of the T cell receptor (TCR) with antigen/MHC proteins must be accompanied by conjugation of T cell counterreceptors (CD28 or CTLA-4) with costimulatory molecules CD80 or CD86 (B7-1 or B7-2) on antigen presenting cells (APC) to avert T cell anergy, and to provide essential signals for T cell activation and cytokine production. However, T cells and APC express changing patterns of counterreceptors and costimulatory molecules during the immune response. To determine the involvement of CD80 and CD86 in costimulation of T cell cytokine production, T cells were incubated with peritoneal exudate macrophages, which express CD80 and CD86, and stimulated in vitro for 48 or 72 hrs with anti-CD3 in the presence or absence of blocking antibody to CD80 or CD86. Alternatively, enriched anti-CD3 stimulated T cells were costimulated with antibody to CD28 and CTLA-4. Production of T cell IL-2, IL-4, and IL-5 was depressed in the presence of anti-CD86 but not anti-CD80. Production of IFN-γ was significantly blocked by either anti-CD80 and anti-CD86. Anti-CD28 was a potent costimulator of IFN-γ and IL-2 production, but a less potent costimulator of IL-4 and IL-5 production. The data suggest that T cell counterreceptors and APC costimulatory molecules act with varying efficacies at stimulating production of T cell cytokines.     

Hydatidiform Mole Macromolecules Inhibit Interleukin-2-Mediated Murine Lymphocyte Proliferation In Vitro

ABSTRACT: Macromolecules extracted from hydatidiform mole trophoblast inhibit mitogen-induced lymphocyte proliferation. To characterize the mechanism of this immunomodulation, we determined the effects of hydatidiform mole vesicle fluid (HMF) and tissue extracts (HME) on lymphokine function in vitro. Utilization of interleukin-1 (IL-1) and interleukin-2 (IL-2) were determined by using a lymphoma cell line (LBRM-33-1A5) and a murine T cell line (CTLL2), respectively. HMF suppressed (P < .05) IL-2-dependent CTLL2 cell proliferation at 500 (36.4% of controls) and 50 (74.9% of controls) μg/ml. HME also suppressed CTLL2 proliferation (P < .05) at 500 (46.0% of controls), 100 (67.2% of controls), 50 (71.5% of controls), and 10 (85.4% of controls) μg/culture ml. In contrast, HMF exhibited no effect on IL-1-stimulated LBRM-33-1A5 production of IL-2. However, 500 μg/ml of HME inhibited (P < .05) IL-2 production (63.0% of controls) in the IL-1 utilization assay. This suppressive effect was probably due to a carry over of HME from the LBRM-33-1A5 culture to the target cells (CTLL2) used to measure IL-2 production. Molecular weight chromatography of an HME sample eluted an IL-2 inhibitor in a low molecular weight (35–50 kd) and high molecular weight (> 250 kd) fraction. These data suggest that one way in which macromolecules derived from hydatidiform mole could interfere with in vitro immunologic responses is by modulating interleukin-2 function.