体外预处理对猪胰岛细胞免疫原性的影响  

Effects of pretreatments of swine islets on immunogenicity of islet graft

作　　者：董维平[1] 王煜非[1] 彭永德[1] 

机构地区：[1]上海交通大学附属第一人民医院糖尿病研究室内分泌科,200080

出　　处：《中华细胞与干细胞杂志（电子版）》2012年第2期8-12,共5页Chinese Journal of Cell and Stem Cell（Electronic Edition）

摘　　要：目的比较多种体外预处理对胰岛细胞免疫原性的影响,筛选既能降低胰岛细胞免疫原性、又不影响胰岛细胞功能的预处理方法。方法新生猪胰岛随机分为:A组(24℃培养5 d组)、B组(液氮冷冻保存组)、C组(中波紫外激光照射组)、D组[主要组织相容性复合体(MHC)Ⅱ类抗原抗体预处理组]和E组(37℃培养5 d组,对照组)。以胰岛-淋巴细胞混合培养的刺激指数(SI)、SLA-DR单抗阳性细胞数和淋巴细胞计数比较各组胰岛的免疫原性;以胰岛素释放试验比较各组胰岛的功能。以上各指标五组间的检测结果差异用单因素方差分析,两两比较采用LSD法。结果预处理各组ICCs的SI值分别为2.2±0.8、1.8±0.6、2.0±1.5、2.1±1.0和7.4±2.3,各组与对照组比较,差异有统计学意义(F=30.398,P<0.01);预处理各组的ICCs中SLA-DR阳性细胞数,分别为2.9﹪±0.4﹪、2.3﹪±0.8﹪、2.4﹪±0.3﹪、2.0﹪±0.8﹪和12.5﹪±5.9﹪,各组与对照组比较,差异有统计学意义(F=103.154,P=0.01);预处理各组ICCs内的淋巴样细胞数,分别为29.0±4.1、27.0±4.0、28.0±2.8、2.6±4.0和111.0±14.0,各组与对照组比较,差异有统计学意义(F=276.212,P<0.01);经预处理的ICCs与对照组的胰岛素释放试验结果,高糖浓度下各组分别为(832±157)、(592±126)、(783±109)、(872±105)及(773±141)pmol/L,各组与对照组比较,差异有统计学意义(F=6.918,P=0.000),而低糖组和释放指数,各组差异无统计学意义(F=0.629,P=0.644;F=0.686,P=0.991)。结论本组资料结果表明,胰岛移植物经短期24℃培养、冷冻保存、中波紫外线激光照射和抗MHCⅡ类抗原单抗预处理,明显降低其免疫原性,同时对胰岛功能无明显影响。Objective To evaluate the effects of different pretreatment methods on the immunogenicity and function of the islet graft.Methods Neonatal pig islets were freshly isolated,and randomly divided into 5 groups： Group A（cultured at 24℃ for 5 days）;Group B（cryopreservated in liquid nitrogen）;Group C（irradiated by medium wave UV laser）;Group D（pretreated by MHCⅡantibody） and control group（cultured at 37℃ for 5 days）.The immunogenicity of islets in each group was analyzed by islet-lymphocyte mixed culture stimulation index（SI）,the numbers of SLA-DR-positive cells and lymphoid cells.The function of islets in each group was evaluated by insulin release test.Comparison among the 5 groups was made with single factor analysis of variance.LSD test was used for comparison between 2 groups.Results SI was 2.2 ± 0.8,1.8 ± 0.6,2.0 ± 1.5,2.1 ± 1.0 and 7.4 ± 2.3 for Groups A,B,C,D and E respectively and was significantly different from control group（F =30.398,P 0.01）.Percentages of SLA-DR positive cells were 2.9﹪± 0.4﹪,2.3﹪± 0.8﹪,2.4﹪± 0.3﹪,2.0﹪± 0.8﹪ and 12.5﹪± 5.9﹪ for the 5 groups respectively,and was significantly different from control group（F = 103.154,P = 0.01）;The numbers of lymphocyte-like cells were 29.0 ± 4.1,27.0 ± 4.0,28.0 ± 2.8,2.6 ± 4.0,111.0± 14.0 for the 5 groups respectively,and was significantly different from control group（F = 276.212,P 0.01）;The re was no significant difference in insulin release test between the pretreatment groups and the control group.Insulin released under high glucose stimulus was（832 ± 157）,（592 ± 126）,（783 ± 109）,（872 ± 105）and 773 ± 141）pmol/L for the 5 groups respectively,and was significantly higher than the control group.（F = 6.918,P = 0.000）,There was no significant difference of stimulation index of low glucose,（F = 0.629,P = 0.644;F = 0.686,P = 0.991）.Conclusion Pretreatments by short-term culture at 24℃,cryopreservation in liquid nitrogen,irradiation by medium wave UV la

关 键 词：胰岛细胞 预处理 免疫原性 猪 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]