同基因背景小鼠核蛋白诱导狼疮鼠模型

背景：自发性狼疮鼠模型不能对基因以外其他的致病因素进行研究。目的：以同基因背景Balb/c小鼠核蛋白免疫小鼠后诱导狼疮鼠模型。方法：选取4～6周SPF级Balb/c小鼠30只,等分为3组。V1组肌肉注射提取的同系Balb/c小鼠核蛋白,间隔3周免疫1次,共免疫4次;V2组注射等体积PBS;V3组为正常对照。检测小鼠末次免疫后3周的24h尿蛋白、血清抗核抗体、抗双链DNA抗体、小鼠肾脏直接免疫荧光。结果与结论：V1组小鼠24h尿蛋白、血清抗双链DNA抗体、抗核抗体均明显高于V2组、V3组,且V1组小鼠肾小球有免疫球蛋白G免疫复合物沉积,可见肾小球轮廓,V2组、V3组未见肾小球轮廓,只见非特异性的微弱荧光。说明以同基因背景Balb/c小鼠核蛋白免疫Balb/c小鼠能够成功诱导狼疮鼠模型。     

脐带间充质干细胞移植对系统性红斑狼疮免疫系统的影响

背景:目前有关脐带间充质干细胞对系统性红斑狼疮的免疫抑制作用及对各个脏器的影响报道较少。目的:探讨脐带间充质干细胞治疗MRL/lpr狼疮鼠的疗效及对免疫系统和各个脏器的影响。方法:MRL/lpr狼疮小鼠随机分为对照组、环磷酰胺组、环磷酰胺+脐带间充质干细胞组、脐带间充质干细胞组。对照组给以生理盐水,其他各组分别给予相应的治疗。采用考马斯亮蓝法检测24h尿蛋白定量;酶联免疫吸附法检测白细胞介素10、白细胞介素17、干扰素γ、PDGF、抗ds-DNA抗体;间接免疫荧光法检测抗核抗体水平;常规病理苏木精-伊红染色观察小鼠的肾脏病理改变肾脏病理改变;免疫组织化学法检测间充质干细胞表面标志CD44、CD105在肾脏、肺脏、脾脏中的表达。结果与结论:①环磷酰胺+脐带间充质干细胞组干扰素γ、白细胞介素17、白细胞介素10水平与对照组相比明显降低(P<0.05)。②32周时环磷酰胺+脐带间充质干细胞组和脐带间充质干细胞组尿蛋白定量低于环磷酰胺组和对照组(P<0.01)。③32周时环磷酰胺+脐带间充质干细胞组抗ds-DNA抗体水平与对照组相比明显降低。④脐带间充质干细胞组肾小球硬化及炎性细胞浸润程度均较对照组为轻。⑤32周时肾脏、肺脏、脾脏中间充质干细胞表面标志CD44、CD105表达为阴性。提示,应用脐带间充质干细胞移植治疗MRL/lpr狼疮鼠可以显著降低狼疮活动指标,对狼疮肾炎具有治疗作用,同时可以通过降低促炎因子的水平发挥免疫抑制作用。     

脐带间充质干细胞移植对系统性红斑狼疮免疫系统的影响

背景：目前有关脐带间充质干细胞对系统性红斑狼疮的免疫抑制作用及对各个脏器的影响报道较少。目的：探讨脐带间充质干细胞治疗MRL/lpr狼疮鼠的疗效及对免疫系统和各个脏器的影响。方法：MRL/lpr狼疮小鼠随机分为对照组、环磷酰胺组、环磷酰胺＋脐带间充质干细胞组、脐带间充质干细胞组。对照组给以生理盐水,其他各组分别给予相应的治疗。采用考马斯亮蓝法检测24h尿蛋白定量;酶联免疫吸附法检测白细胞介素10、白细胞介素17、干扰素γ、PDGF、抗ds-DNA抗体;间接免疫荧光法检测抗核抗体水平;常规病理苏木精-伊红染色观察小鼠的肾脏病理改变肾脏病理改变;免疫组织化学法检测间充质干细胞表面标志CD44、CD105在肾脏、肺脏、脾脏中的表达。结果与结论：①环磷酰胺＋脐带间充质干细胞组干扰素γ、白细胞介素17、白细胞介素10水平与对照组相比明显降低（P〈0.05）。②32周时环磷酰胺＋脐带间充质干细胞组和脐带间充质干细胞组尿蛋白定量低于环磷酰胺组和对照组（P〈0.01）。③32周时环磷酰胺＋脐带间充质干细胞组抗ds-DNA抗体水平与对照组相比明显降低。④脐带间充质干细胞组肾小球硬化及炎性细胞浸润程度均较对照组为轻。⑤32周时肾脏、肺脏、脾脏中间充质干细胞表面标志CD44、CD105表达为阴性。提示,应用脐带间充质干细胞移植治疗MRL/lpr狼疮鼠可以显著降低狼疮活动指标,对狼疮肾炎具有治疗作用,同时可以通过降低促炎因子的水平发挥免疫抑制作用。     

载脂蛋白E基因缺乏对C57BL/6小鼠狼疮早期抗体产生的影响

目的 用两种不同狼疮造模方法研究载脂蛋白E基因缺乏(apoE-/-)对C57BL/6(B6)小鼠系统性红斑狼疮发病早期抗体产生的影响.方法 将36只apoE-/- C57BL/6雌性小鼠分为降植烷对照组,降植烷模型组,脂多糖对照组,脂多糖模型组;同时将34只普通B6雌性小鼠亦分为降植烷对照组,降植烷模型组,脂多糖对照组,脂多糖模型组;降植烷对照组一次性腹腔注射生理盐水0.5 mL,降植烷模型组一次性腹腔注射降植烷0.5 mL,脂多糖对照组1周2次注射生理盐水,每次0.2 mL,脂多糖模型组1周2次注射脂多糖溶液0.2 mL,浓度为250 mg/L;3周后采血测定血清中抗核抗体(ANA)、抗dsDNA抗体、抗可溶性抗原(ENA)抗体水平,并比较各组间的差异.结果 ANA各模型组均较对照组升高,且apoE-/-的降植烷模型组较普通B6降植烷模型组升高;抗dsDNA抗体仅apoE-/-的降植烷模型组较对照组升高;抗ENA抗体apoE-/-的脂多糖模型组较对照组及普通B6脂多糖模型组升高.结论 在降植烷诱导的模型早期,apoE-/-可促进雌性B6小鼠ANA及抗dsDNA抗体的产生;在脂多糖诱导的模型早期,apoE-/-可促进雌性B6小鼠抗ENA抗体的产生.     

钙蛋白酶抑制蛋白基因过表达Balb/C小鼠的繁育及鉴定

目的:建立钙蛋白酶抑制蛋白(calpastatin,CAST)基因过表达Balb/C小鼠的培育方法,并探讨碱性裂解液提取基因组DNA在子代鼠基因型鉴定中的价值。方法:将雄性C57BL/6-CAST~(+/-)转基因小鼠与雌性Balb/C野生型小鼠杂交,将雄性CAST~(+/-)杂合子代小鼠与雌性Balb/C野生型子代小鼠杂交,连续杂交至消除C57BL/6小鼠遗传背景。取子代小鼠鼠尾组织,采用碱性裂解液提取基因组DNA,PCR法扩增CAST基因片段,琼脂糖凝胶电泳鉴定结果。采用柯萨奇B3病毒(CVB3)感染Balb/C-CAST~(-/-)小鼠以建立急性病毒性心肌炎小鼠模型。结果:C57BL/6-CAST~(+/-)转基因小鼠与雌性Balb/C野生型小鼠连续杂交10代可消除C57BL/6小鼠遗传背景。采用碱性裂解液提取的基因组DNA数量、纯度能够满足后续实验需要,且产物大小与目的片段一致。成功建立Balb/C-CAST~(-/-)基因型小鼠急性病毒性心肌炎模型。结论:成功建立Balb/CCAST基因过表达小鼠,为后续研究奠定了基础;碱性裂解液提取基因组DNA简便、高效,值得推广。     

白细胞介素-6在ALD-DNA诱导的SLE模型小鼠SLE中的作用机制研究

目的探讨白细胞介素(IL)-6对ALD-DNA诱导的系统性红斑狼疮(SLE)模型小鼠SLE中的作用机制。方法在健康的雌性C57BL/6小鼠体内用活化淋巴细胞来源的DNA(ALD-DNA)免疫,从而诱导小鼠的SLE,未免疫的小鼠被用作对照。检测了疾病相关的一些发病指标,包括抗双链DNA抗体,尿蛋白和肾脏的病理学改变。结果 IL-6KO基因敲除的小鼠能抵抗ALD-DNA诱导的小鼠SLE模型。在IL-6KO免疫的小鼠中,CD4~+T细胞的活化状态低于野生型的免疫小鼠。胞内细胞因子染色结果表明,IL-6KO免疫的小鼠体内Foxp3的表达高于野生型免疫的小鼠。结论在ALD-DNA诱导的SLE模型中,IL-6可以抑制Treg细胞的分化,从而促进疾病的发展。     

丙泊酚通过抑制小鼠海马神经元突触释放组织纤溶酶原激活物引起神经毒性损伤

目的：建立钙蛋白酶抑制蛋白（calpastatin,CAST）基因过表达Balb/C小鼠的培育方法,并探讨碱性裂解液提取基因组DNA在子代鼠基因型鉴定中的价值。方法：将雄性C57BL/6-CAST+/-转基因小鼠与雌性Balb/C野生型小鼠杂交,将雄性CAST+/-杂合子代小鼠与雌性Balb/C野生型子代小鼠杂交,连续杂交至消除C57BL/6小鼠遗传背景。取子代小鼠鼠尾组织,采用碱性裂解液提取基因组DNA,PCR法扩增CAST基因片段,琼脂糖凝胶电泳鉴定结果。采用柯萨奇B3病毒（CVB3）感染Balb/C-CAST-/-小鼠以建立急性病毒性心肌炎小鼠模型。结果：C57BL/6 CAST+/-转基因小鼠与雌性Balb/C野生型小鼠连续杂交10代可消除C57BL/6小鼠遗传背景。采用碱性裂解液提取的基因组DNA数量、纯度能够满足后续实验需要,且产物大小与目的片段一致。成功建立Balb/C-CAST-/-基因型小鼠急性病毒性心肌炎模型。结论：成功建立Balb/C-CAST基因过表达小鼠,为后续研究奠定了基础;碱性裂解液提取基因组DNA简便、高效,值得推广。     

三氧化二砷对MRL-lpr狼疮肾炎小鼠淋巴细胞凋亡及相关基因的影响

目的通过检测淋巴细胞凋亡比例、凋亡相关基因表达的变化情况,探讨三氧化二砷对MRL-lpr狼疮肾炎小鼠淋巴细胞凋亡的影响。方法14只MRL-lpr小鼠,随机分为两组,实验组用三氧化二砷隔日腹腔注射,对照组用生理盐水,疗程结束后检测两组小鼠抗双链DNA抗体水平、用流式细胞仪检测两组小鼠脾脏淋巴细胞凋亡情况、逆转录-聚合酶链反应检测基因caspase-3的表达水平及蛋白印记法测bcl-2的蛋白表达。结果实验组抗双链DNA抗体水平显著低于对照组,脾脏淋巴细胞凋亡较对照组明显增多,caspase-3水平亦明显增高,而bcl-2表达减少。结论三氧化二砷能够诱导MRL-lpr狼疮肾炎小鼠自身反应性淋巴细胞发生凋亡,其作用机制可能与激活caspase家族成员、抑制bcl-2等自身基因表达有关。     

一组抗HCV核壳蛋白的单克隆抗体的制备及鉴定

目的 获取HCV 核壳蛋白抗原结构新的信息。方法 以基因重组的HCV 核壳蛋白区抗原免疫 Balb/c 小鼠,利用小鼠杂交瘤技术。结果 建立了42株分泌抗 HCV区单克隆抗体(单抗)的杂交瘤细胞株。其抗体类别均为IgG。经免疫印迹提示本组单抗与HCV 核壳蛋白有反应。结论 IFA(免疫荧光)证实本组单抗能识别天然 HCV核壳蛋白抗原决定簇。     

T细胞受体β独特型抗原决定簇DNA疫苗诱导抗淋巴瘤抗体的研究

目的 观察T细胞受体（TCR）β不同独特型抗原决定簇DNA疫苗诱导BALB/c小鼠抗人类CEM淋巴瘤细胞免疫反应的情况。方法 RT-PCR扩增CEM细胞特异性重排的TCRβ基因片段，克隆至真核表达载体pcDNA3中，然后以此为模板，扩增编码不同抗原决定簇的基因片段到pcDNA3中作为DNA疫苗。肌肉注射法免疫小鼠，间接免疫荧光法检测小鼠血清中抗体生成情况。结果 CEM细胞TCRβ共包含5个抗原决定簇，其DNA疫苗免疫的6只小鼠全部产生了特异性抗独特型抗体（最高滴度1：480）；含4个抗原决定簇的DNA疫苗免疫的6只小鼠中有4只产生了特异性抗独特型抗体，但滴度均较低（最高滴度1：80）；仅含2个抗原决定簇的DNA疫苗未能使小鼠产生特异性抗独特型抗体。结论 包含TCRβV区全长，共有5个抗原决定簇的独特型DNA疫苗可以诱导小鼠产生抗淋巴瘤细胞的独特型特异性抗体；含有4个抗原决定簇的DNA疫苗仍可诱导特异性抗体的生成，但这种反应较弱，而仅含有2个抗原决定簇的DNA疫苗则不能诱导小鼠产生抗淋巴瘤的特异性抗体。     

HSV-1感染后小胶质细胞炎性因子分泌特点研究

目的：比较单纯疱疹病毒I型（HSV-1）感染小鼠和BV2细胞后,小胶质细胞分泌炎性因子的特征,探讨BV2细胞可否作为小鼠HSV-1性脑炎模型的小胶质细胞功能的模拟研究对象。方法：24只Ba1b／c小鼠分为正常组、HSV-1组、黄芪组及地塞米松组（激素组）各6只;BV2细胞分为同样4组。HSV-1组,黄芪组及激素组小鼠颅内注射HSV-1制造小鼠HSV-1性脑炎（HSE）模型,BV2细胞接种HSV-1制造细胞模型。造模成功后,黄芪组及激素组分别用黄芪及激素作用前后,RT-PCR法检查炎性因子（IL-2、IL-4、IL-10、TNF-α及iNOS）mRNA浓度及细胞的变化。结果：BV2细胞与模型小鼠在病毒感染及相关药物作用前后,炎性因子具有一致的变化趋势。结论：BV2细胞和Balb／c小鼠脑组织具有相似的免疫反应特性,可作为研究单纯疱疹病毒性脑炎小鼠模型中小胶质细胞功能的模拟对象。     

新型抗狂犬病病毒核蛋白抗体进行RFFIT检测效果评估

目的评价自主研发的荧光标记抗狂犬病病毒核蛋白单克隆抗体用于快速荧光灶抑制试验(RFFIT)的效果。方法将狂犬病病毒核蛋白的一段线性表位肽与KLH耦联后免疫Balb/c小鼠制备的高效价单克隆抗体抗-RVNP-Mcl,经浓缩、纯化、异硫氰酸荧光素(FITC)标记用于RFFIT检测,以Millipore公司DFA5100为对照试剂,对200例人血清进行平行RFFIT检测,将结果进行统计学分析。结果两种抗体检测结果的线性相关分析显示相关系数(r)=0.980;两种抗体检测定性结果一致率为96.83%,卡方检验显示两种抗体检测定性结果差异无统计学意义(χ2=0.098,P>0.05),发生定性结果不一致的6例(3.00%)血清两种抗体检测结果均小于1 IU/mL;68例两种抗体检测结果均小于或等于1 IU/mL的血清样品中有6例(8.82%)发生结果定性不一致,其余样品均未发生不一致的情况。结论使用荧光标记抗-RVNP-Mcl与DFA5100进行RFFIT检测时结果高度一致,荧光标记抗-RVNP-Mc1可供RFFIT检测使用。     

Impact of blood transfusion and burn injury on microbial translocation and bacterial survival

The role of the immune system in microbial translocation must be clarified. In these studies, the effect of blood transfusion-related immunosuppression on translocation was investigated in a burn animal model previously known to increase the gut's permeability to 14C-radiolabeled Escherichia coli. In a first experiment, Balb/c mice underwent transfusion (T) with 0.2 mL per mouse of allogeneic C3H/HeJ mouse blood 5 days prior to undergoing 30-percent burn injury (B) and simultaneous gavage (G) with 10(9) E. coli bacteria labeled with 14C. An additional six groups of Balb/c mice underwent different combinations of T, B, and G procedures (TG, BG, TB, T, B, G). Survival rate was recorded for all groups on Day 10. This experiment suggested that B and T, to a lesser extent, were the factors affecting survival, although the combination of T, B, and G clearly showed a synergistic effect on mortality. In a second experiment, 18 Balb/c mice belonging to TBG, BG, TG, and G groups were sacrificed 1, 4, and 24 hours after burn or gavage. The residual radioactivity and the percentage of viable bacteria were computed for mesenteric lymph nodes, spleen, liver, lungs, blood, and peritoneal fluid. Statistical analysis of the radionuclide counts recognized B as the only variable able to enhance the magnitude of 14C E. coli translocation. The percentage of viable bacteria showed that T and, more moderately, B were the factors leading to the failure of bacterial clearance in the tissues.     

Donor WBCs can persist and transiently mediate immunologic function in a murine transfusion model: effects of irradiation, storage, and histocompatibility

BACKGROUND: Donor WBCs are responsible for numerous transfusion complications, but little is known concerning the natural history of their clearance following transfusion or of their function in the recipient's circulation. A murine transfusion model was developed to investigate the effects of blood component characteristics and histocompatibility on donor WBC survival kinetics and function. STUDY DESIGN AND METHODS: To investigate the effects of storage and irradiation, fresh whole blood and blood stored for 1, 2, and 6 weeks at 4 degrees C, all from male C57b (H2K(b)) mice, was transfused to female Balb/c (H2K(d)) mice. To study the effect of histocompatibility, blood was also transfused from C57b mice to Balb/c, FVB, C3H, and SW (outbred) mice. To investigate the xenogeneic setting, blood from humans, rats, and rabbits was transfused to Balb/c mice. Samples were collected weekly after transfusion, and the donor WBCs were analyzed, targeting the Y-chromosome with quantitative PCR. To investigate donor WBC function, dinitrochlorobenzene (DNCB) sensitivity was induced in donor and recipient mice, and the transfusion recipients were observed for hypersensitivity to DNCB. RESULTS: Donor WBCs had reduced in vivo survival equivalent to their period of storage ex vivo at 4 degrees C. Irradiation of donor blood produced no observable difference in donor WBC survival. Allogeneic male donor WBCs persisted (100-<1 cell/microL) in female Balb/c recipient mice blood over 6 weeks. Donor WBC survival kinetics displayed an early MHC-dependent phase, which was followed by a more rapid phase that was not influenced by donor-recipient MHC differences. All donor WBCs were cleared within 24 to 48 hours. DNCB sensitivity was passed through transfusion, where it was transiently expressed in naive recipients. CONCLUSION: The clearance of donor WBCs in the murine transfusion model is much slower than that in humans. Allogeneic donor WBC clearance may be biphasic, involving MHC-dependent as well as MHC-independent mechanisms. DNCB sensitivity can be transferred transiently to a naive recipient.     

Detection of antioxidant enzyme activities in renal tissues of early stage IgA nephropathy in ddY mice

The purpose of this study was to determine the antioxidant enzyme activities in renal tissues of early stage ddY mice, an animal model for primary IgA nephropathy. Eight- and 40-week-old ddY female mice and normal healthy Balb/c female mice were used in this study. The levels of Cu/Zn-SOD, Mn-SOD, and GSH-PX activities in the renal cortex were significantly higher in 40-week-old ddY mice than in Balb/c control mice of the same age; no change of catalase activity was observed. There were no significant differences in the levels of Cu/Zn-SOD, MT-SOD, GSH-PX, and catalase activities between the ddY mice and Balb/c mice at 8 weeks of age. Urinary protein was slightly higher in 40-week-old ddY mice. IgA or C3 was deposited at low levels in the glomerular mesangial areas of 8-week-old ddY mice. Marked depositions of IgA and C3 extended from the glomerular mesangial areas to the capillary walls of 40-week-old ddY mice. Expansion of glomerular mesangial matrices and mild mesangial cell proliferation was observed in 40-week-old ddY mice. Antioxidant enzyme activities in the renal cortex were already increased in the early stage IgA nephropathy in 40-week-old ddY mice. These findings suggest that measurements of antioxidant enzyme activities in the renal cortex of 40-week-old ddY mice was useful for evaluation of the pathogenesis of renal involvement in the early stage of IgA nephropathy. © 1996 Wiley-Liss, Inc.     

应用Tail-cuff技术改良小鼠颈部异位心脏移植模型

背景:小鼠颈部心脏移植模型是研究移植缺血再灌注损伤和排斥反应的理想模型,由于显微血管吻合的技术问题,限制了小鼠颈部异位心脏移植模型在医学研究中的广泛应用.目的:应用Tail-cuff技术改良小鼠颈部异位心脏移植模型.方法:选用Balb/c→Balb/c小鼠进行同系心脏移植,C57BL/6→Balb/c小鼠进行同种心脏移植.将24G和22G静脉内导管制作成带有尾巴的套管(Tail-cuff),在受体血管准备时,Tail-cuff通过右颈外静脉或右颈总动脉近心端后,用显微血管夹一起固定其近心端和Tail-cuff管的尾巴,翻转血管并固定后分别与供心肺动脉主干和升主动脉连接固定.结果与结论:共36例次正式实验,12例次同系移植,24例次同种移植,移植成功率为100%.应用改良技术后总操作时间和心肌缺血时间分别缩短为(49.6±7.4)min和(28.8±4.2)min,尤其是血管翻转和与供心连接时间明显缩短.结果提示应用Tail-cuff技术建立小鼠颈部异位心脏移植模型显示出一定的优越性,是一种理想的方法.     

Assessment of the role of antibiotics and enterococcal virulence factors in a mouse model of extraintestinal translocation

OBJECTIVE: To study the relative contribution of antibiotics and bacterial virulence factors in the process of translocation of Enterococcus faecalis from the gut to extraintestinal organs. DESIGN: Prospective controlled animal study. SETTING: Animal experimental laboratory at a university medical center. SUBJECTS: Fifty-two female Balb/c mice. INTERVENTIONS: We developed a mouse model to study the translocation of Enterococcus faecalis from the intestinal tract. Balb/c mice received sterile drinking water or antibiotic combinations to deplete their indigenous intestinal microflora. The animals subsequently were fed genetically engineered enterococci expressing different combinations of the putative enterococcal virulence factors aggregation substance and binding substance. Animals were killed, and their livers, spleens, and mesenteric lymph nodes were aseptically removed and cultured along with fecal samples for enumeration of bacteria. MEASUREMENTS AND MAIN RESULTS: All animals were colonized with the test strains at 2-6 x 109 colony forming units/g of feces; in the antibiotic-treated animals, feces were free from anaerobes and Enterobacteriaceae. In animals fed the identical bacterial mutant, the colony counts in mesenteric lymph nodes were significantly lower in mice not treated with antibiotics than in those treated with antibiotics (p =.016). Multigroup analysis of variance revealed no significant differences of the translocation frequencies for the different mutant strains; however, the differences were statistically significant for all groups receiving antibiotics vs. the group not receiving antibiotics (p <.05-.01). There was a trend (although not statistically significant) for a higher proportion of positive cultures from either spleen or liver in mice that had enterococci recovered from their mesenteric lymph nodes (28%) relative to those that did not have enterococci isolated from the lymph nodes (12%; rate ratio 2.39, p =.30 by logistic regression analysis). CONCLUSIONS: Oral antibiotics can select for extraintestinal translocation of Enterococcus faecalis, and neither aggregation substance nor binding substance seems to be required for this process. The experiments encourage further exploration of host and microbial factors contributing to translocation and may provide a better understanding of the pathogenesis of enterococcal infections in patients in intensive care units.     

胸腺内注射异基因抗原诱导特异性神经移植耐受的实验

背景:主要组织相容性复合物抗原不匹配是发生移植排斥的主要原因,诱导免疫耐受是防止器官移植排斥的理想途径,在胸腺内通过阴性选择可获得对自身耐受的T细胞,胸腺内异基因抗原注射是否可获得对该抗原的免疫耐受?目的:观察胸腺内注射异基因抗原在同种异基因神经移植免疫耐受中的作用。设计:对照观察实验。单位:哈尔滨医科大学免疫学教研室,哈尔滨医科大学第四临床医院骨科。材料:供体鼠C57BL/6(H-2b)30只,雄性,6～8周龄,体质量18～22g,由黑龙江省兽医研究所提供;受体鼠Balb/c(H-2d)60只,雌性,6～8周龄,体质量18～22g,由北京实验动物中心提供。主要组织相容性复合物(H-2b)抗原由哈尔滨医科大学免疫教研室制备,蛋白浓度为4.4g/L。方法:实验于2002-06/11在哈尔滨医科大学免疫教研室完成。将受体鼠Balb/c随机分为4组:胸腺内注射组、同系同基因移植组、异体神经移植组、免疫抑制剂组。自供体鼠C57BL/6(H-2b)的脾细胞中提取MHC抗原,注入受体鼠Balb/c(H-2d)胸腺内,两周后移植供体坐骨神经。移植后3周做单向混合淋巴细胞培养,诱导迟发型超敏反应。主要观察指标:各组混合淋巴细胞反应、诱导迟发型超敏反应的差异。结果:纳入供体鼠C57BL/6(H-2b)30只和受体鼠Balb/c(H-2d)60只,全部进入结果分析。①混合淋巴细胞反应结果:同系同基因移植组、胸腺内注射组和免疫抑制剂组细胞增殖均明显低于异体神经移植组[(546.1±75.1),(2668.3±533.8),(3101.3±429.1),(4312.3±534.1)min-1,P<0.05]。②诱导迟发型超敏反应结果:同系同基因移植组、胸腺内注射组和免疫抑制剂组两侧足垫的厚度差均明显低于异体神经移植组[(41.1±3.7),(72.1±5.1),(57.6±11.3),(86.2±13.2)μm,P<0.05]。结论:胸腺内注射异基因H-2b抗原可以诱导特异性神经移植耐受。     

QUANTITATIVE INVESTIGATIONS OF IDIOTYPIC ANTIBODIES : VI. IDIOTYPIC SPECIFICITY AS A POTENTIAL GENETIC MARKER FOR THE VARIABLE REGIONS OF MOUSE IMMUNOGLOBULIN POLYPEPTIDE CHAINS

Antisera were prepared in rabbits against anti-p-azobenzoate antibodies of an A/J and a BALB/c mouse and anti-p-azophenylarsonate antibodies of an A/J mouse. After appropriate absorption the antisera reacted with the anti-hapten antibody of the donor mouse but, by sensitive quantitative tests, not at all with other components of the hyperimmune serum or with preimmune serum of the donor mouse. The absorbed antiserum therefore appeared to be specific for idiotypic determinants. Nearly all idiotypic specificities identified in the serum of the donor were also present in the serum of other mice of the same strain, immunized against the same hapten group, but not in mice immunized with a different hapten. In each case the antibodies of the donor mouse reacted most effectively on a weight basis with antiidiotypic antiserum. Cross-reactions were observed among different strains of mice but homologous anti-bodies reacted most effectively with antiidiotypic antisera. C57/BL and DBA antisera contained very low concentrations of specificities present in the A/J and BALB/c antibody populations; antibodies of A/J and BALB/c antisera are more closely related to one another. The results indicate that idiotypic specificity may provide a genetic marker for the variable regions of immunoglobulin polypeptide chains.