人耳软骨移植抗原性及脱细胞处理对其的影响

目的 研究人耳软骨的移植抗原性及脱细胞处理对其的影响。方法 用低渗的TritonX—100为主的方法脱去人耳软骨细胞，然后通过抗人主要组织相容性复合体—I(MHC—I)免疫组织化学染色，与单个核细胞共同培养来研究人耳软骨的移植抗原性及脱细胞处理对其的影响。结果 软骨细胞脑浆抗人MHC—I为阳性，抗原的分布以细胞核周边的细胞质为主。细胞外基质抗人MHC—I为阴性。而经过脱细胞处理后的人耳软骨则呈抗人MHC—I阴性。单个核细胞与软骨共同培养时则显示出较高DNA合成能力，与对照组比较有统计学意义(P＜0．05)。单个核细胞与脱细胞处理的软骨共同培养后的DNA合成能力较低，与单纯的单个核细胞培养展示出新的DNA合成能力，但无显著差异(P＞0．05)。单个核细胞与正常人耳软骨共同培养的过程中，可见移向软骨的单个核细胞随时间增加而增加。相反，单个核细胞与脱细胞处理的软骨共同培养后，却少见单个核细胞移向脱细胞处理的软骨(P＜0．01)。结论 软骨具有移植抗原性；以低渗的TritonX—100为主的脱细胞方法可以有效的去除移植物的抗原性，从而避免同种异体组织移植排斥反应的发生。     

Evaluation of resorption and biocompatibility of collagen hemostats in the spinal epidural space

Background contextCollagen hemostats have different characteristics depending on their properties and configuration. In vivo serial evaluation of local reactions because of placement of hemostats in the epidural space has not been reported.PurposeThis study compared the resorption and biocompatibility of two types of collagen hemostats placed in the epidural space.Study designThis in vivo study used experimental animals to evaluate collagen hemostats that were placed in the epidural space.MethodsA ligamentum flavum resection model was created in Japanese white rabbits (n=65). A microfibrillar collagen hemostat (MCH group, n=5), cotton-type collagen hemostat (CCH group, n=5) that was chemically cross-linked, or no hemostat (control group, n=4) was placed in the spinal epidural space. For histologic evaluation, each group was euthanized 1, 2, 4, and 8 weeks postoperatively (PO), and hematoxylin-eosin and immunohistochemical (IHC) staining for inflammatory cytokines (tumor necrosis factor [TNF]-α, interleukin [IL]-6), cyclooxygenase (COX)-2, and macrophages (CD68) was performed. To evaluate exudate accumulation and the degree of inflammation in the epidural space, magnetic resonance imaging at 7.04 T was serially performed in each group (n=3) under anesthesia and sedation.ResultsThe collagen hemostats in both groups were reabsorbed at 4 weeks PO. In the MCH group, there was inflammatory cell infiltration and granuloma formation around the hemostat, TNF-α–positive cells were seen up to 1 week, and IL-6–, COX-2–, and CD68-positive cells were seen at all evaluation times. In the CCH group, no inflammatory cell infiltration around the hemostat was observed, and IHC staining showed no positive cells at 4 weeks PO and later. T2*-weighted MR images showed significantly higher mean signal intensity of the epidural space in the MCH group than in the CCH group but only at 1 week PO (p<.05).ConclusionsResorption of both hemostats was similar. In the MCH group, there was intense tissue inflammation around the hemostatic material, and MR images showed high signal intensity because of exudate accumulation in the epidural space. This indicated a strong foreign-body reaction to the MCH, thus demonstrating a difference in biocompatibility with the CCH.     

两种方法筛选噬菌体肽库所获日本血吸虫抗原模拟表位的抗原性比较

目的比较两种不同方法筛选噬菌体肽库所获得的日本血吸虫抗原模拟表位的抗原性.方法以日本血吸虫病患者血清免疫球蛋白(Sj-Ig)作为靶分子, 分别以噬菌体12肽库(SjA)和经过蚯蚓免疫兔血清免疫球蛋白(L-Ig)筛选一轮后的噬菌体12肽库(SjB)为原肽库,进行3轮吸附-洗脱-扩增免疫筛选.随机挑取SjA和SjB蓝色噬菌斑各24个, 扩增后用ELISA方法检测其抗原性.结果 24个SjA克隆中有6个可以与Sj-Ig特异结合, 其中A491 nm值较高的2个克隆具有较好的特异性和敏感性;24个SjB克隆中有10个可以与Sj-Ig特异结合, 其中A491 nm值较高的5个克隆具有较好的抗原性.比较SjA 和SjB两组间克隆的抗原性,结果显示SjB的目的克隆具有更好的特异性和敏感性.结论从SjA 和SjB中均筛选到了对日本血吸虫病具有较高潜在诊断价值的抗原模拟表位,提示以异源性抗体结合后的肽库作为原肽库来筛选寄生虫抗原模拟表位是可行的,同时也为从噬菌体肽库中筛选抗原模拟表位提供了一个新的实验方法.     

日本血吸虫虫卵中毛蚴抗原的融合表达及抗原性分析

目的 重组表达制备日本血吸虫虫卵中毛蚴抗原（SjMP10）。 方法 根据日本血吸虫虫卵中毛蚴抗原SjMP10分子的读框序列设计合成1对引物，扩增SjMP10 DNA片段并克隆到表达载体pGEX-4T-3中。转化BL21(DE3)后，诱导表达谷胱甘肽转移酶-虫卵中毛蚴抗原10（GST-SjMP10）融合蛋白。采用洗脱法制备GST-SjMP10融合蛋白，应用免疫印迹法（Western blotting)和淋巴细胞增殖试验进行重组蛋白的抗原性分析。 结果 SjMP10基因克隆到表达质粒pGEX-4T-3后，经异丙基-β-D-硫代半乳糖苷(IPTG)诱导能表达出Mr 39 000的GST-SjMP10融合蛋白，经电洗脱法纯化的上述重组融合蛋白可被血吸虫感染兔血清所识别，并能刺激血吸虫感染鼠脾淋巴细胞增殖。 结论 日本血吸虫虫卵中毛蚴抗原SjMP10融合表达获得成功。     

The effect of polyamines on the binding of anti-DNA antibodies from patients with SLE and normal human subjects

Antibodies to DNA (anti-DNA) are the serological hallmark of systemic lupus erythematosus (SLE). To elucidate specificity further, the effect of polyamines on the binding of anti-DNA antibodies from patients with lupus was tested by ELISA to calf thymus (CT) DNA; we also assessed the binding of plasmas of patients and normal human subjects (NHS) to Micrococcus luteus (MC) DNA. As these studies showed, spermine can dose-dependently inhibit SLE anti-DNA binding to CT DNA and can promote dissociation of preformed immune complexes. With MC DNA as antigen, spermine failed to inhibit the NHS anti-DNA binding. Studies using plasmas adsorbed to a CT DNA cellulose affinity indicated that SLE plasmas are mixtures of anti-DNA that differ in inhibition by spermine and binding to conserved and non-conserved determinants. Together, these studies demonstrate that spermine can influence the binding of anti-DNA autoantibodies and may contribute to the antigenicity of DNA.     

日本血吸虫Calpain蛋白免疫原性研究

目的：探讨日本血吸虫疫苗候选分子Calpain蛋白的抗原性及其所诱导的免疫应答类型。方法：用Genetyx—MAC9．0分析Calpain肽链的亲水性，在亲水性区域以9个氨基酸为肽链单位的固相重叠肽链库(每相邻两个肽链之间仅有一个氨基酸不同)被Auto—spot Robot制备，用Dot—ELISA在合成的肽链库上筛选B细胞表位，含有B细胞表位的肽链免疫BALB／c小鼠，ELISA鉴别特异性IgG抗体亚类的产生。结果：由758个氨基酸组成的日本血吸虫Calpain蛋白含有两个主要的亲水区，其存在区域分别在氨基酸229～375和555～613；测定合成的肽链文库，在亲水区内筛选出4个B细胞的表位，分别是氨基酸234～251(YAHISGGT)；290～297(CLMGCSIH)；338～346(PWGDSHEW)：358～364(AWCDGA PQ)；与对照组比较，免疫组鼠血清中IgG1、IgG2a和IgG2b特异性抗体有显著性意义的上升。结论：日本血吸虫Calpain是一个具有免疫原性的蛋白，能够激发宿主产生高水平的免疫球蛋白，提示Calpain可以发展成为日本血吸虫病的诊断抗原和日本血吸虫疫苗候选分子。     

肾综合征出血热病毒PS-6株和JR-C-1株抗原性的比较研究

目的 比较肾综合征出血热（HFRS）汉坦病毒（HV）PS-6株和JR-C-1株的抗原性，选择制备疫苗的毒种，方法 应用空斑减少中和试验（PRNT）和McAb间接免疫荧光试验（IFAT），检测PS-6株和JR-C-1株HFRSHV的抗原性。结果 PS-6和JR-C-1均为Ⅰ型HV。两株病毒的免疫血清对17株国内外不同来源的HV的中和反应和76-118株汉滩病毒（HTHV的免疫血清基本相似。但对一些病毒的中和反应效价两有差异。与多株McAb反应结果显示，两株病毒间存在抗原决定簇的差异。结论 PS-6和JR-C-1的抗原性与76-118株HTHV基本相同，PS-6和JR-C-1之间存在抗原性和抗原决定簇的差异。两合用抗原性会有互补作用。     

Pre-clinical immunogenicity of human papillomavirus alpha-7 and alpha-9 major capsid proteins

Human papillomavirus (HPV) vaccines confer protection against the oncogenic genotypes HPV16 and HPV18 through the generation of type-specific neutralizing antibodies raised against the constituent virus-like particles (VLP) based upon the major capsid proteins (L1) of these genotypes. The vaccines also confer a degree of cross-protection against some genetically related types from the Alpha-9 (HPV16-like: HPV31, HPV33, HPV35, HPV52, HPV58) and Alpha-7 (HPV18-like: HPV39, HPV45, HPV59, HPV68) species groups. The mechanism of cross-protection is unclear but may involve antibodies capable of recognizing shared inter-genotype epitopes. The relationship(s) between the genetic and antigenic diversity of the L1 protein, particularly for non-vaccine genotypes, is poorly understood.     

Decrease of blood type antigenicity over the long-term after ABO-incompatible kidney transplantation

Background

Few studies have investigated the changes in the antigenicities of the transplanted organs after transplantation.

Methods

We examined, by immunohistochemical assay, the changes in expression of the blood-type antigens on the transplanted kidneys over the long-term after ABO-incompatible kidney transplantation with A- or B-antigen incompatibility (blood type A to B and B to A). The subjects were six patients, including one case with graft loss, who had received ABO-incompatible kidney allografts more than ten years previously. As normal controls, four cases of ABO-compatible transplantation during the same period, including two recipient/donor pairs each with blood group A1 and blood group B were enrolled.

Results

Expression of the blood-type A or B antigens decreased gradually to 91.8% during the first three months after transplantation, 85.8% during the first five years, 64.1% during the first ten years, and 57.6% over ten years after transplantation on average in ABO-incompatible transplant recipients. In one patient with graft loss due to severe antibody-mediated rejection, the donor's type B blood-type antigens on the transplanted graft had changed but partially to the recipient's blood-type A antigen by 2582 days after the transplantation, suggestive of the occurrence of blood-type chimerism on the endothelium. In ABO-compatible transplant recipients, such changes in expression were not observed. The average percentage of blood-type antigen-positive vessels at more than ten years after the renal transplantation was 99.8%.

Conclusions

Decrease in the expression of the donor's blood-type antigen on the endothelium of the graft has been considered as one of the mechanisms underlying the accommodation occurring over the long-term after ABO-incompatible kidney transplantation. On the other hand, establishment of antigenic chimerism on the graft endothelium could be one of the hallmarks of the immunological reaction associated with antibody-mediated rejection.     

鼠疫耶尔森菌外膜蛋白抗原性研究

目的 建立分析鼠疫耶尔森菌外膜蛋白抗原性的简便方法. 方法 采用DNAstarProtean和EMBOSS网络分析平台综合预测鼠疫耶尔森菌外膜蛋白YopE、YopH、YopT、YopJ、YopM的免疫原性,并通过蛋白质芯片技术对上述外膜蛋白的免疫原性进行验证. 结果 计算机综合预测分析显示外膜蛋白YopE抗原...