CD80、CD86抗体对耗竭性T细胞效应功能的影响

目的以CD80、CD86抗体作为激动剂、脾细胞为研究对象,观察CD80、CD86抗体对耗竭性T细胞效应功能的影响。方法将42只8周龄雌性HLA-A11DR1转基因小鼠随机分为7组各6只,分别在8、11、14周龄时,A^F组臀部皮下接种磷脂酰肌醇蛋白聚糖3(GPC3)多肽,G组接种等体积无热源PBS;A^E组在24周龄和F、G组在17周龄时脱颈处死,取脾脏并制成脾细胞悬液;A、B组分别滴加20、40μg/m L抗鼠CD80抗体和10μg/m L GPC3多肽,C、D组分别滴加20、40μg/m L抗鼠CD86抗体和10μg/m L GPC3多肽,E、F、G组仅滴加10μg/m L GPC3多肽。孵育18 h后,采用酶联斑点分析仪检测干扰素γ(IFN-γ),以此判断T细胞效应功能。结果 A^G组脾细胞IFN-γ阳性斑点数分别为(80.61±48.91)、(207.67±60.41)、(1.67±0.97)、(1.33±0.49)、(2.33±1.53)、(38.17±5.18)、(2.33±1.53)个。其中,F组高于E、G组(P均<0.01),E、G组间比较无统计学差异;A组>B组>C、D、E组(P均<0.01),C、D、E组间比较差异无统计学意义;而且,IFN-γ阳性斑点数随着CD80抗体浓度升高而增加,二者呈正相关(r=0.760 5,P<0.01)。结论 CD80抗体能够刺激耗竭性T细胞恢复产生细胞因子,且呈剂量依赖的动力效应;而CD86抗体未能检测到该功能。

The immunophenotypic changes and clinical effectiveness after treatment of cancer patients with infusion of human peripheral blood lymphocytes stimulated by anti-CD28 and anti-CD80 monoclonal antibodies in combination with radiotherapy and chemotherapy

To investigate the changes on the immunopbenotypes and the clinical effects of treatment of the late cancer patients with infusion of human peripheral blood lymphocytes stimulated by anti-CD28 and anti-CD80 monoclonal antibodies in combination with radiotherapy and chemotherapy, 42 patients with late cancers were collected for study, among which 22 patients were treated with infusion of stimulated lymphocytes in combination with radiotherapy and chemotherapy. The immunological treatment procedure was given twice per week, and one course of treatment consisted of 8 times of giving infusion of lymphocytes. Another 20 patients were selected for control group, in which only radiotherapy and chemotherapy were given without lymphocyte infusions. Flow cytometry was used to examine the immunophenotypes and the clinical symptoms were observed before and after treatments. It was found that the numbers of the CD3^ ＋ , CD4^＋ cells increased, while those of the CD8 ^＋ cells decreased, with an increase of CD4/CD8 radios, but no significant difference existed in case of 22 patients treated with lymphocyte infusion as well as with radiotherapy and chemotherapy. Fifteen patients out of these 22 cases （68.18%）, the immunophenotypes changed obviously with increased numbers of CD3^ ＋ , CD4^ ＋ cells in comparison with those before treatment, and the number of CD95^ ＋ cells was increased after treatment. The PS value in this group of patients decreased after treatment. In comparison with 20 cases in the control group, the immunophenotypes showed no differences before and after treatment. While the PS value decreased obviously. Seven out of the 22 cases （31.83 % ） treated with lymphocyte infusions as well as with radiotherapy and chemotherapy illustrated no major changes in their i mmunophenotypes, compared with the situation before treatment, but the PS value also decreased. In case of treatment with lymphocyte infusions in combination with radiotherapy and chemotherapy, the alteration of phenotypes was reversely correlated with the changes of clinical grades. Although there were 7 cases showing no major alterations of the immunological phenotypes, but their correlation was still evident. In the control group, neither alteration of immunophenotypes nor changes in clinical grades was found. It is concluded that immunotherapy in combination with radiotherapy and chemotherapy can relieve the side effects induced by radiotherapy and chemotherapy and also enhance the therapeutic efforts.

免疫因子对慢性免疫性血小板减少性紫癜骨髓巨核祖细胞体外生长的影响

目的 :探讨慢性免疫性血小板减少性紫癜 (CITP)骨髓巨核祖细胞生长成熟与免疫因素的关系。方法 :通过巨核祖细胞体外培养 ,观察CITP骨髓巨核祖细胞生长和成熟情况 ,并观察抗CD80抗体及干扰素α 2b(IFN α 2b)对CITP和对照组巨核祖细胞生长和成熟的影响 ,并与对照组比较。结果 :①CITP组的各种集落数(CFU MK、BFU MK、mCFU MK)均低于对照组 ,两组间集落数的均值比较差异有统计学意义 (P <0 .0 5 )。CITP组的直径和面积均低于对照组 ,两组间直径和面积均值比较差异有统计学意义 (P <0 .0 5 )。②加入抗CD80抗体的CITP的巨核祖细胞集落生成数明显增多 (P <0 .0 5 )。③IFN α 2b对两组的总集落均有抑制 ,CITP比对照组的巨核祖细胞集落生成受抑制程度低 (P <0 .0 5 )。结论 :CITP巨核祖细胞的集落生成和成熟度较低。抗CD80抗体和IFN α

CD80mAb协同imDC诱导同种异体大鼠胰十二指肠移植免疫耐受

目的 观察共刺激分子阻断剂CD80单克隆抗体（CD80mAb）在协同未成熟树突细胞（imDC）诱导同种异体大鼠胰十二指肠移植免疫耐受中的作用。方法 建立糖尿病大鼠胰十二指肠移植动物模型；4E5杂交瘤细胞株BABIMC小鼠腹腔注射，抽取腹水，分离纯化后获得CD80mAb；分离供体大鼠骨髓来源DC细胞前体，经GM—CSF、IL-4体外刺激后。再加入IL-10共培养，鉴定为imDC；移植前7d，将2×10^6imDC经静脉途径注射至受体体内，同时分别给予生理盐水1ml、CD80mAb5mg连续14d。结果 四组受体大鼠移植后中位生存时间分别为12．7d、32．4d、50．2d、92．0d，实验组存活时间明显延长；组织学观察发现移植后7dCD80mAb＋imDC组移植物形态尚完整，淋巴细胞浸润减少；混合淋巴细胞反应证实移植后7dCD80mAb＋imDC组供受体间呈低反应性。结论 共刺激分子阻断剂CD80mAb能够协同imDC诱导受体T细胞对移植物的免疫耐受，降低宿主对移植物的急、慢性排斥反应，延长移植物的存活时间。