Characterization of gp 50, a major glycoprotein present in rat brain synaptic membranes, with a monoclonal antibody

Several cell lines secreting monoclonal antibodies (Mabs) against a major forebrain synaptic membrane (SM) glycoprotein, gp 50, have been raised. Western blots show that the Mabs react with a polypeptide doublet of Mrs 49 and 45 kDa. These polypeptides exist solely in a concanavalin A (Con A) binding form. Removal of the Con A receptors by digestion with endo-beta-N-acetylglucosaminidase H (endo H) lowers the Mrs of the glycoprotein doublet to 36.5 and 34 kDa. Western blots of 2D polyacrylamide gels indicate that gp 50 exists in several isoforms. Solid phase radioimmunoassay (RIA) and Western blots of brain subcellular fractions show the antigenic material to be concentrated in the SM fraction, but to be present in much lower amounts in synaptic junctions and postsynaptic densities. Gp 50 appears to be brain specific. Regional distribution studies show that it is present in all brain regions but is two-fold concentrated in cerebellum, brainstem and midbrain compared to forebrain. Immunocytochemical studies of several brain regions show that gp 50-like immunoreactivity is neuron specific and is concentrated in selected neuronal species, particularly granule cells. In both cerebellar and hippocampal granule cells gp 50-like immunoreactivity is localized in the perikarya and primary dendrites. Though immunocytochemistry did not show staining of synaptic regions this may be due to masking of the reactive epitope. The results are discussed in terms of the molecular properties of gp 50 and its subcellular localization in brain tissue.     

间接免疫荧光法检测特异性IgG抗体对流行性出血热诊断意义

本文报告用间接免疫荧光法检测特异性IgG抗体对流行性出血热的诊断意义。53例不同病期的出血热患者的血清,其间接荧光抗体的阳性率为49/53(占92.45％)。有77.36％的患者血清,检出IgG抗体的时间是在发病后的1～2周。血清中IgG的滴度与疾病的严重性无密切相关。另25例非EHF患者,其血清特异性IgG抗体均为阴性。可见检测特异性IgG抗体对EHF的诊断颇为有用。     

An epitope shared by central nervous system myelin and peripheral blood macrophages

Lewis rats were immunized with a homogenate of human spinal cord. Splenocytes from the immunized rats were fused with cells from the SP2/0-Ag14 cell line to form hybrids that were subsequently screened immunohistochemically for secretion of antibodies against myelin. Thirty hybrids secreting anti-myelin antibodies were cloned. One secreted antibody (774) that immunohistochemically stained central nervous system (CNS) myelin but not peripheral nervous system (PNS) myelin also bound to the surface of peripheral blood macrophages. Hence we have identified an epitope that is shared by peripheral blood macrophages and CNS myelin.     

白细胞介素-15对骨髓增生异常综合征骨髓造血干/祖细胞增殖的影响

目的：观察白细胞介素(IL-15)对体外培养的骨髓增生异常综合征(MDS)患者CD34^ 细胞增殖作用。方法：应用单克隆抗体免疫磁珠分离系统提取MDS患者CD34^ 细胞，以加IL-15组为实验组，不加IL-15组为对照组，进行液体和甲基纤维素半固体集落培养，计算培养后细胞数和CFU—E、BFU—E、CFU—GM、CFU—GEMM等集落数，并用MTT比色法检测IL-15对MDS患者CD34^ 细胞增殖的抑制作用，流式细胞术检测上述培养细胞周期的变异情况。结果：11例MDS对象平均CD34^ 细胞比例、回收率、纯度和富集倍数均达要求，MTT比色法检测IL-15对CD34^ 细胞的增殖作用呈最佳浓度效应，最佳浓度为20μg／L，细胞增殖抑制最低峰值时间为8d。用0μg／L IL-15(对照组)和20μg／L IL-15(实验组)作用MDS CD34^ 细胞，计数显示培养细胞最大增殖倍数和集落形成比率实验组均较对照组明显增加，IL-15作用后各细胞周期G1、S、G2期比例有明显改变，与对照组比较，差异有统计学意义。结论：IL-15对MDS CD34^ 细胞有促增殖效应，与其它造血生长因子具有协同作用，对MDS治疗可能有一定的应用前景。     

Kidney Transplantation in Patients with Antibodies against Donor HLA Class II

The immunologic risk associated with donor-specific antibodies (DSA) against Class II human leukocyte antigens (HLA) in kidney transplant (KTx) recipients is unclear. The aim of this study was to determine the outcome of KTx when DSA was detected only against HLA Class II. To isolate the impact of anti-Class II DSA, we retrospectively analyzed 12 KTx recipients who at baseline had a positive B-cell flow cytometric crossmatch (FXM) and a negative T-cell FXM. Using alloantibody specification analysis, 58.3% (7/12) had DSA against donor Class II and 41.7% had no demonstrable DSA. Biopsy-proven AMR occurred in 57% (4/7) in the Class II(+) group and 0% in the Class II(-) group (p > 0.05). Peritubular capillaries stained positive for C4d in 86% (6/7) of the Class II(+) patients and in 40% (2/5) of the Class II(-) patients (p > 0.05). One patient in the Class II(+) group lost their graft at 3 months to accelerated transplant glomerulopathy, while all other grafts were functioning 3-37 months posttransplant despite the persistence of anti-Class II DSA. We conclude that KTx recipients with clearly defined anti-Class II DSA are at risk for humoral rejection suggesting that desensitization and/or close posttransplant monitoring may be needed to prevent AMR.     

P物质抑制剂辣椒素和bFGF抗体对增生性瘢痕成纤维细胞增殖协同抑制作用的实验研究

目的：探讨P物质抑制剂-辣椒素及bFGF抗体单独或联合使用对体外培养的人体增生性瘢痕成纤维细胞增殖的影响。方法：体外培养12例增生性瘢痕组织的成纤维细胞,分别加入不同浓度的辣椒素或/和bFGF抗体,培养24h后观察细胞形态学变化,并用MTT法检测细胞增殖抑制率。结果：所有不同浓度的辣椒素（2.5,5,20,40,60,80mg/L）均能使细胞皱缩、坏死,抑制细胞增殖,其抑制作用随剂量增大而增加。中高浓度bFGF抗体（40,80mg/L）也可促使细胞皱缩、坏死,抑制细胞增殖,其抑制作用也随剂量增大而增加。辣椒素与bFGF抗体联合应用对成纤维细胞抑制作用较单独使用效果更显著（P﹤0.05）。结论：辣椒素及bFGF抗体可抑制增生性瘢痕成纤维细胞增殖,其抑制作用随浓度增加而增加,两者联合使用有协调作用,本研究结果可能在增生性瘢痕的治疗提供新的思路和手段。     

抗神经元抗体对副肿瘤综合征早期诊断的临床意义

目的 探讨抗神经元抗体抗-Hu、抗-Yo、抗-Ri抗体的检测与副肿瘤综合征早期诊断的关系。方法 采用蛋白免疫印迹分析(Western Blot)法对12例确诊为副肿瘤综合症(PNS)的患者血清及32例非PNS血清进行检测。结果 12例确诊患者血清中检测到一项或多项抗体阳性;PNS患者血清中抗-Hu、抗-Yo、抗-Ri抗体的阳性率高于非PNS对照组,差异有极显著性意义(P<0.01);正常对照组血清抗-Hu,抗-Yo,抗-Ri抗体全部为阴性。结论 抗神经元抗体抗-Hu、抗-Yo、抗-Ri抗体可作为PNS早期诊断及确诊的指标。     

TSo联合IFN-γ鼻内免疫小鼠诱导的弓形虫IgA抗体特异性免疫应答

目的研究速殖子超声裂解物(TSo)和IFN-γ鼻内免疫小鼠经口感染弓形虫速殖子后,血清、小肠液和粪便IgA抗体分泌的动态变化。方法将6～7周龄BALB/c小鼠100只随机分为4组,每组25只,分别用10μl缓冲液PBS、20μgTSo、500UIFN-γ和20μgTSo 500UIFN-γ鼻内免疫小鼠。用RH株弓形虫速殖子4×104个/只灌胃攻击,分别于攻击后第7、10、13、16、19天处死小鼠,收集血清、小肠液及粪便,ELISA法测定IgA含量。结果各组血清IgA抗体水平在攻击后第10天达到峰值,TSo IFN-γ组血清IgA抗体含量高于其他各组。小肠液和粪便IgA抗体含量在攻击后第13天达到峰值,在实验期各时点TSo IFN-γ组小肠液和粪便IgA抗体含量高于其他组。小肠液与粪便IgA抗体水平呈正相关。结论TSo或IFN-γ或TSo联合IFN-γ鼻内免疫小鼠可诱导黏膜免疫,产生高水平的弓形虫特异性IgA抗体。TSo联合IFN-γ鼻内免疫优于单独免疫,免疫小鼠肠道产生大量SIgA,发挥抗虫作用。鼻黏膜免疫是一种安全有效的免疫接种途径。     

17β-雌二醇对子宫内膜异位症患者在位子宫内膜间质细胞β-catenin mRNA和蛋白表达的影响

目的研究17β-雌二醇(17β-E2)对子宫内膜异位症(内异症)患者在位子宫内膜间质细胞β-catenin mRNA和蛋白表达的影响,探讨Wnt/β-catenin信号通路在介导雌激素促进内异症发生发展的作用。方法体外分离培养内异症患者在位子宫内膜间质细胞。用不同浓度17β-E2处理子宫内膜间质细胞48 h;此后选用10-10mol/L 17β-E2处理子宫内膜间质细胞12、24和48 h,逆转录聚合酶链反应(RT-PCR)和免疫印迹法(Western blotting)检测17β-E2处理前后子宫内膜间质细胞β-catenin mRNA和蛋白的表达水平。同法分析雌激素受体拮抗剂ICI182,780(10-6mol/L)对17β-E2促进β-catenin mRNA和蛋白表达的影响。免疫组织化学染色观察17β-E2作用后β-catenin在子宫内膜间质细胞中的定位。结果17β-E2能明显促进内异症患者在位子宫内膜间质细胞β-catenin mRNA和蛋白的表达,并呈剂量和时间依赖性,于10-10mol/L作用48 h最明显。雌激素受体拮抗剂ICI182,780能明显抑制17β-E2对子宫内膜间质细胞β-catenin mRNA和蛋白的表达。免疫组织化学染色发现17β-E2能促进β-catenin在子宫内膜间质细胞核内的表达。结论雌激素可能通过激活Wnt/β-catenin信号通路促进内异症在位子宫内膜的异位种植。