利用独特型——抗独特型理论检测Graves病患者血清中异常免疫球蛋白

根据体外放射分析的原理及独特型——抗独特型理论,利用抗TSH抗体作为结合剂,放射性碘标记的金黄色葡萄球菌A蛋白(~(125)I-SPA)作为定量的示踪剂,建立一种新的检测 Graves病(GD)患者血清中的异常免疫球蛋白(aIgG)的方法,进行了有关方法学的初步研究,探索了TSHAb与aIgG反应的最适反应条件,并与以神经节苷脂为结合剂的酶联免疫吸附分析法(ELISA)作了比较.探讨了aIgG与甲状腺刺激性免疫球蛋白(TSI)的关系.结果显示:TSHAb与GD患者血清中aIgG在体外能够发生特异性结合,而与正常人血清中的IgG、系统性红班狼疮(SLE)患者IgG及糖尿病(DM)患者IgG不发生结合反应.在以aIgG指数作为评价指标时,29名正常人血清aIgG指数为1.06±0.17,其正常范围上限为1.39,在以大于1.39作为阳性时,72名不同临床时期GD患者,初发组阳性率为83%(n=24),临床缓解组阳性率为12%(n=25),复发组阳性率为82%(n=23).正常人血清aIgG指数值与GD初发患者及GD复发患者相比,差异具有极显著性(P<0.001).我们认为本法能较好检出该类aIgG,可用于反映GD患者免疫状况,鉴别诊断及预测治疗缓解后复发的有用指标.     

胆结石患者血清和胆汁幽门螺杆菌相关蛋白免疫印迹检测

目的：研究幽门螺杆菌（HP）感染与胆石形成的关系。方法：采用聚合酶链免疫吸附法（ELISA）对胆结石患者的血清及胆汁免疫球蛋白IgG,IgA,IgM标本进行检测，对血清学及胆汁IgG均阳性患者的胆汁，采用免疫印迹法（Western blot)进行HP感染相关蛋白检测。结果：胆结石组50例患者中血清学及胆汁IgG均阳性者29例（58.0%)，另取非胆结石组13例作为对照，胆结石组与非胆结石组血清及胆汁HP免疫印迹检测，均可检出4种主要HP感染相关蛋白，但胆结石组血清及胆汁中UreA检出率较非胆结石组高。结论：胆汁中存在多种HP感染相关蛋白，并可能参与胆结石的形成。     

Identification of the major allergenic proteins from silkworm moth (Bombyx mori) involved in respiratory allergic diseases

Introduction and ObjectivesMoths are a significant source of indoor and outdoor aeroallergens. High prevalence of IgE-mediated sensitization was demonstrated in a group of patients with allergic respiratory diseases. There are no studies on adult stage of these moth species allergens involved in allergic respiratory reactions - the aim of this study.Material and Methods36 participants were included in an experimental study, submitted to skin prick test with Bombyx mori wing extract and six other common allergens, as well as Western blot analysis with incubated nitrocellulose membrane impregnated with silkworm moth extract and human IgE-antibody. The participants were divided into 3 groups: 1) 21 allergic patients whose skin prick test was positive to Bombyx mori wing extract, 2) eight allergic patients whose skin prick test was positive to mite and negative to Bombyx mori extract 3) seven negative non-allergic subjects.ResultsAmong the 21 participants from group 1, 19 serum samples reacted to Bombyx mori extract by Western blot. All of them reacted to a protein at 80 kDa and five other proteins (66, 50, 45, 37 and 30 kDa) were identified in more than 50% of the individuals tested, considered as major allergenic proteins. Sera from seven out of eight patients sensitized to house dust mite demonstrated IgE-reactivity to Bombyx mori extract by Western blot analysis. Serum samples from healthy participants did not react at all.ConclusionSix major reactive proteins by immunoblot analysis from moth’s wings sensitized patients can be potential allergens. The one at 80 kDa is the major protein, seen in all IgE-reactive patients from group 1 and in none from group 2, yet to be identified. Future studies should be conducted to better characterize these proteins.     

Effects of d-penicillamine on mononuclear cells in vitro

Summary d-penicillamine (d-pen) inhibited pokeweed mitogen-induced plaque-forming cell (PFC) response in a dose-dependent manner. This inhibition was irreversible as preincubation for a few hours with the drug followed by washes still caused suppression of the PFC response. Pretreatment of the different mononuclear cell populations with d-pen for short periods (2–24 h) showed that both macrophages (Mo) and B lymphocytes were affected by the drug. By contrast T cells were resistant. Mo appears to be more susceptible to d-pen than B cells, and in the case of drug-treated Mo, the response was restored completely with the addition of 20% fresh Mo. Our results show that d-pen, without exogenous Cu²⁺, inhibits the polyclonal immunoglobulin secretion by human mononuclear cells in vitro due to a strong effect on both Mo and B cells. This may explain the decrease in serum immunoglobulin levels seen in patients with rheumatoid arthritis undergoing this therapy.     

Hodgkin's disease developing after spontaneous remission of chronic lymphocytic leukemia

We present a 71-year-old patient with chronic lymphocytic leukemia diagnosed 27 years ago. Initially, the disease was staged as Rai II and the patient suffered from secondary immunoglobulin deficiency. Nevertheless, no treatment was necessary at that time. Because of disease progression a single course of chemotherapy was given in 1984. During the following year there was a constant decline of the WBC, accompanied by normalization of the immunoglobulins; both have remained stable ever since that time. However, there was still residual bone marrow infiltration, indicating persisting CLL. In 1993 cervical lymphadenopathy occurred with acute onset. A diagnostic lymphadenectomy revealed Hodgkin's disease of the nodular-sclerosing subtype. The patient was staged as II–III according to the Ann Arbor Classification and underwent radiation therapy. Cytogenetic examination of the bone marrow revealed a normal karyotype with an inversion of chromosome 9. This case demonstrates the rare coincidence of two lymphoproliferative disorders in the same patient. The clinical course and the immunologic findings of this patient are presented, together with a review of the literature.     

天津某医院严重急性呼吸综合征暴发期间血清流行病学调查

目的 研究本院严重SARS暴发流行后，疫源地不同人群血清SARS冠状病毒（SARS-CoV）抗体水平及其产生和变化规律；探讨人群SARS-CoV隐性感染和IgG抗体的保护作用。方法 采用血清流行病学法，联合运用ELISA和免疫荧光试验（IFA），调查疫情暴发流行期间及流行后，疫源地内非SARS人群血清抗体水平变化；定性研究SARS患者病后6周内IgM、IgG抗体产生和变化规律；动态观察SARS患者康复期82周内IgG抗体水平变化规律。结果 各100例疫源地一般人群和非疫源地对照人群血清SARS-CoV IgG抗体ELISA抽样检测均为阴性；487例SARS高危人群血清SARS-CoV IgG抗体ELISA检测阳性率为0．41％，经IFA复核后均为阴性；疫源地内非SARS人群IgG抗体A值水平在疫情流行后高于流行期间，差异有统计学意义（P〈0．05）；SARS患者病后1～6周血清抗体IgM和IgG的阳性检出率分别为7．7％、40．0％、57．8％、88．2％、76．6％、57．1％和0、23．1％、48．4％、65．4％、77．8％、100．0％；SARS患者康复期血清IgG抗体A值逐渐增高，病后第22周达到高峰，此后缓慢下降，第82周时仍维持较高水平，下降趋势减缓。结论 SARS可能不存在隐性感染者；绝大多数SARS患者在急性期或恢复早期的血清中存在IgM抗体，其出现较早，消失也较快，抗体IgG出现稍晚，但在血清中存在时间较长，达到高峰后消退缓慢，提示该抗体可能具有保护作用。     

基因扩增技术检测急慢性B—淋巴细胞白血病免疫球蛋白基因重排

应用聚合酶链反应(PCR)扩增IgH重排产生的CDR-Ⅱ序列,检测23例急慢性B-淋巴细胞白血病患者。20例发生IgH重排,其中4例出现两种基因扩增产物。6例完全缓解的患者,3例检出异常重排的IgH基因,并先后复发。本文应用Southern blot法和PCR进行了对照性研究。     

A Rheumatoid Factor Specific Mimotope Identified by a Peptide Display Library

We screened a 10 amino acid peptide display library in filamentous phage with B'20, a monoclonal high affinity IgM rheumatoid factor (RF) expressing the Vk111a-dependent 4C9 idiotype. Using direct and indirect selection techniques, 12 B'20 reactive peptides were identified, 9 of which belonged to one of two motifs. Binding of B'20 to phage-bearing peptides was inhibited by both IgG and 4C9 antiidiotype. Synthetic peptides corresponding to the two motifs inhibited the Fc binding of a low avidity IgA B'20 construct. Purified IgM from 6/8 RF-positive RA patients and 8/11 monoclonal RFs with Vk111-encoded light chains bound to the phage, whereas none of the four monoclonal RFs with Vk1 or Vk11 encoded light chains bound. Phage binding appeared to be RF specific. Three 4C9 positive/RF negative cell lines from RA patients did not bind to phage nor did three B'20 mutants that had lost RF specificity, whereas two mutants that retained RF specificity also retained phage binding. We propose that there is a common epitope(s) recognized by Vk11 I encoded RFs that is mimicked by the structure of these peptides. Such mimotopes might be exploited to design novel agents that interfere with autoantibody binding.     

Inhibition of Fas-mediated Apoptosis by Antigen: Implications for Lymphomagenesis

Apoptotic deletion of expanded B cell populations is essential in avoidance of autoimmune disease and immune regulation of some B cell malignancies. The role of CD4+ T cells in B cell apoptosis is evident from the high incidence of B cell tumors and autoimmunity in patients with T cell diseases such as the acquired immune deficiency syndrome (AIDS). We have previously demonstrated that in Epstein-Barr Virus (EBV) negative Burkitt's lymphoma (BL), a tumor derived from proliferating centroblasts of the germinal center, the malignant lymphocytes can be induced to express Fas (CD95) by ligation of CD40 at the B cell surface. Upon CD40 engagement, BL cells are sensitized to T-cell derived death signals provided by Fas ligand (FasL, CD95L). HBL-3 is a cell line derived from an AIDS-related BL in which the tumor IgM binds the human erythrocyte "i" antigen. To determine whether Fas-mediated apoptosis of BL cells is reduced in the context of antigen to which the tumor IgM binds, we stimulated HBL-3 cells with CD40 ligand (CD40L, CD154) in the presence and absence of human erythrocytes expressing the "i" antigen, and measured Fas-mediated apoptosis upon exposure to an agonistic anti-Fas antibody. We observed that HBL-3 cells were sensitized to Fas-mediated death by exposure to CD40L. When i+ RBCs were present, Fas-mediated apoptosis in HBL-3 cells was reduced by greater than 30%. In contrast, there was no reduction in Fas-mediated apoptosis in the presence of i &#109 (I+) RBCs. These findings demonstrate that Fas-mediated deletion of BL cells is inhibited upon surface IgM engagement by antigen for which the malignant clone has affinity.     

Anti-idiotypes to Oxidized LDL Antibodies in Intravenous Immunoglobulin Preparations--Possible Immunomodulation of Atherosclerosis

Objective : The aim of this study was to examine whether intravenous immunoglobulin (IVIg) preparations contain anti-oxLDL and anti-anti-oxLDL antibodies. Background : Oxidized low-density lipoprotein (oxLDL) is one of the major players in atherogenesis. IVIg can reduce atherosclerosis in experimental animal models. Methods : Six commercial IVIg preparations were tested for the presence of anti-oxLDL antibodies by EIA. Inhibition studies were performed with the different IVIg preparations and IgGs purified from a pool of sera from patients with high anti-oxLDL antibody levels. Absorption assays were carried out to evaluate the presence of anti-idiotypes against anti-oxLDL antibodies in IVIg preparations. Results : IVIg preparations tested had various degrees of reactivity towards oxLDL. Absorption experiments suggested that the reactivity was specific because it could be effectively absorbed by oxLDL and not by an irrelevant antigen PPD. The reactivity was smaller than that observed with the IgG from the pool with high anti-oxLDL antibody levels. Inhibition studies with IVIg demonstrated 20-45% inhibition of anti-oxLDL binding to oxLDL, compared to 76% inhibition by the pool with high anti-oxLDL levels. To investigate the presence of anti-idiotypes against anti-oxLDL antibodies within IVIg, F(ab') 2 fragments of IVIg IgG were used to absorb IgG F(ab') 2 fragments from the pool of sera with high anti-oxLDL levels. The decreased binding to oxLDL of the absorbed supernatants shows that IgG F(ab') 2 fragments of the IVIg preparations had high inhibitory capacities ranging from 65 to 90%. Conclusions : IVIg preparations contain both anti-oxLDL and anti-anti-oxLDL activity. This finding may explain the immunomodulating effect of IVIg in atherosclerosis.