Electrophoretic studies of the conversion products of serum β1C-globulin

By the method of antigen—antibody crossed electrophoresis where the antigens after a first separation in agarose gel are driven into a new antibody containing gel, serum β_1C-globulin and its conversion products were analysed in native serum as well as after addition of EDTA, incubation with immunoprecipitate and hydrazine, and after inactivation at 56°.

It is shown that β_1C-globulin can be converted not only to β_1A-globulin or components with the same mobility as β_1A-globulin but also to additional components, one in the α₂-region, one in the inter-β-region and one in the γ₁-region. α₂- and inter-β-components are seen especially in serum to which EDTA has been added. EDTA did not inhibit the spontaneous conversion of β_1C-globulin.

     

Charge and size of the conversion products of serum β1C-globulin

Using gel filtration on Sephadex G-200 combined with antigen—antibody crossed electrophoresis of the fractions, serum β_1C-globulin and the conversion products present in stored native serum and stored serum to which EDTA had been added, respectively, were analysed. The conversion products in the inter-β- and γ₁-zone demonstrated in EDTA serum after 1 days storage and after hydrazine treatment, respectively, could not be characterized by gel filtration, probably because of their lability. Sephadex filtration did not induce any conversion of β_1C-globulin in serum. It was found that β_1C- and β_1A-globulin left the column in homogeneous peaks in the `7S' peak, the former slightly before the latter. The conversion product obtained in stored EDTA sera and on electrophoresis migrating in the α₂-zone was found in the right extension of the macroglobulin peak. The precipitation line produced by this component was diffuse and differed distinctly from the precipitation lines of the other components. The possibility of some kind of complex is discussed.     

Effect of mercaptans and other agents on the human complement component β1C-globulin

Immunoelectrophoretic analyses and complement titrations of whole human serum show that a number of physical, chemical and immunological agents affect the β_1C/β_1A-globulin system which is considered to represent the third component of complement (C′₃). The transformation from β_1C-globulin to β_1A-globulin is the normal result of ageing, while antigen-antibody complexes, polylysine, hydrazine and cobra venom accelerate this change. In addition, α-globulin fragments of β_1A-globulin appear after interaction of normal sera with antigen-antibody complexes, treatment with cobra venom, and by storage under nitrogen in glass tubes or in polyethylene containers. Similar α-fragments are seen in the aged sera of some patients with glomerulonephritis or renal allografts. With prolonged storage these α-fragments can reform β_1A-globulin. On the other hand, 2-mercaptoethanol and penicillamine produce complete dissolution of β_1C-globulin into rapidly migrating, poorly defined fragments in the α-globulin and albumin regions, and transform β_1A-globulin into a stable β₂-globulin. The reduced fragments of β_1C-globulin, if not alkylated, can subsequently form the β₂-globulin, but not the β_1A-globulin. These results indicate that β_1C-globulin is composed of several subunits, some of which are joined by disulphide bonds, but that the α-globulin fragments seen in some pathological sera are not a result of disulphide bond reduction.     

The relationship between circulating soluble antigen—antibody complexes and the production of chronic glomerulonephritis in the rabbit

Twenty-three rabbits were injected over long periods of time with isotopically labelled bovine serum albumin. Varying injection schedules and primary and secondary responses were studied, but all schedules were designed to preclude the presence of free circulating antibody at any time. All animals became tolerant of injected antigen after a period of 60–80 days. During the intervening period (from 10 to 60 days) large amounts of antigen—antibody complex were shown to be circulating.

Proteinuria was intermittent in all animals. Histological examination of the kidneys from 50 days on showed only very minor evidence of renal damage; no animal developed chronic renal disease.

     

Long-term observation of glomerulonephritis induced by multiple injections with anti-Thy-1 antibody in rats

Multiple Injections with a mouse monoclonal anti-rat Thy-1 antibody (five times, at weekly intervals) induced marked glomerular sclerotic lesions which are characterized by adhesion of glomerular capillaries to Bowman's capsule and persistent proteinuria in rats. Abnormal production of type I collagen and increased accumulation of type IV collagen and flbronectln were observed in these glomeruli. The glomerular expression of mRNA for these matrix components and transforming growth factor-pi (TGF-β1) were markedly increased at 4 days after the last injections with anti-Thy-1 antibody, but decreased to below the levels of control rats at 5 weeks. This may be down-regulation of mRNA In mesangial cells. The glomerular sclerotic lesions were not progressive but the process of glomerular healing seemed to be retarded. The proteinuria and the glomerular adhesion were irreversible.     

Effect of erythrocyte breakdown products on stem cells and erythropoietin formation

In experiments on CBA mice and albino rats the effect of erythrocyte breakdown products (EBP) on the number of colony-forming units (CFU), differentiation of stem cells, and erythropoietin production was studied. After three or four injections of EBP to normal or lethally irradiated (1000 rad) mice, no changes in the number of CFU or in differentiation of the stem cells were observed after transplantation of bone marrow. Daily administration of EBP to mice for 3 days before irradiation (1000 rad) and bone marrow transplantation led to an increase in the number of colonies in the recipients' spleen, mainly on account of colonies of erythroid type. Injection of EBP into the animals did not change the erythropoietic activity of the blood serum. The possible role of EBP in the mechanism of autoregulation of erythropoiesis is discussed.Division of Experimental Pathology and Experimental Diagnosis, Sverdlovsk Scientific-Research Institute of Tuberculosis. Sverdlovsk Scientific-Research Institute of Work Hygiene and Occupational Diseases. Department of Pathological Physiology, Sverdlovsk Medical Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR N. A. Fedorov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 84, No. 8, pp. 143–145, August, 1977.     

Effects of leflunomide on glomerulonephritis induced by antibasement membrane antibody in rats

Leflunomide (HWA 486, a novel isoxazol derivative), shown to have potent immunosuppressant and antiinflammatory effects, was evaluated for its inhibitory and therapeutic effects on the glomerulonephritis induced in rats by rabbit antiserum against rat glomerular basement membrane. Leflunomide was administered orally to rats at 0.5 and 2 mg/kg/day for 20 days from 2 days before injection of the rabbit antiserum and at 2 mg/kg/day for 14 days from 5 days after the antibody injection. The present study consisting of 2 experiments for inhibitory (I) and therapeutic (II) effects of leflunomide revealed the following effects at 2 mg/kg: in experiment I, significant decreases in (a) urinary total protein, (b) plasma total cholesterol and fibrinogen and (c) thymus weight, and decreased incidences of fibrin deposits in Bowman's space, adhesion of the glomerulus to Bowman's capsule and deposition of rat IgG and C₃; and in experiment II, decreases in (a), (b) and (c), though smaller than in experiment I, and decreases deposition of rat C₃. Thus, leflunomide had potent inhibitory and limited therapeutic effects on glomerulonephritis, suggesting that the compound is effective in inhibiting the onset and development of glomerulonephritis.     

Suppressive effects of Sairei-to on monoclonal antibody 1–22–3-induced glomerulonephritis: Analysis of effective components

The effects of treditional Chinese medlclne (Salrel-to) on experimental glomerulonephritls Induced In rats by monodonal antibody (mAb) 1–22–3 lnjectlon was examined. The level of proteinuria in the Sairel-to-treated group was significantly lower than that In the PBS treated group. This suppressive effect was caused by the major component of Sealer-to, Syo-salko-to but not by another component, Gorel-san. The suppressive effect of Syo-salko-to was Identified In Its components ( Bupleuri radix, Pindilae tuber and Zingibers rhizoma ), but not In the other combined components ( Ginseng radix and Zizyphl fructus ). Further study weeled that the suppressive effects of the combined components were mainly derived from Bupleuri radix . It was demonstrated that the actual active Ingredient is probably Salkosaponin-d. Light microscopy revealed that Sairel-to and Its effective components suppressed the proliferation of mesanglal cells and mesanglal matrix expansion. Semi quantitative morphological studies of glomerular lesions on the eighth day showed that Syo-salko-to and Its combined components ( Bupleuri radix, Zinglberis rhizoma and Pinelliae tuber ) suppressed mesanglal matrix expansion significantly compared with phosphate-buffered saline control groups (matrix score: 28.0±19.1 vs 102.3±14.1; 30.9±30.1 vs 102.3±14.1, p<0.005, respectively). It was concluded that Salkosaponln-d, as well as Bupleuri radix , Syo-salko-to and Sairel-to can suppress proteinurla and morphological changes In the rat glomerulonephritls model Induced by mAb 1–22–3.     

SENP1对小鼠B细胞数量及血清免疫球蛋白的影响初探

本研究利用Cre/LoxP系统构建B细胞条件性敲除Senp1基因小鼠模型(CKO),探讨了SUMO特异性蛋白酶1(SENP1)对C57BL/6小鼠B细胞数量以及对血清免疫球蛋白产生的作用。利用流式细胞术检测对照组(WT)及实验组(CKO)小鼠骨髓、脾脏及淋巴结B细胞含量及分布情况,结果显示,Senp1敲除后可显著降低脾脏和淋巴结中B细胞的比例,而对骨髓中的B细胞比例没有影响。利用ELISA方法检测血清中免疫球蛋白IgM、IgG、IgG3及IgG1亚型的水平,结果显示CKO小鼠血清IgG1的含量明显低于WT小鼠,而IgM、IgG及IgG3含量没有显著差别。以上结果提示SENP1在小鼠外周淋巴器官B细胞分布及IgG1产生过程中可能起调控作用。     

Comparative evaluation of three products for the detection of Borrelia burgdorferi antibody in human serum.

Eighty human serum specimens tested concomitantly by immunoblot and an enzyme-linked immunosorbent assay developed jointly at the University of Connecticut School of Medicine and the Connecticut Agricultural Experiment Station were used to evaluate three commercially available diagnostic products for Lyme borreliosis. The sources of the kits were Hillcrest Biologicals, Cypress, Calif.; Whittaker Bioproducts, Walkersville, Md.; and Cambridge Bioscience, Worcester, Mass. When compared with Western blot analysis, the sensitivities and specificities, respectively, for the diagnostic assays were as follows: Hillcrest Biologicals, 93 and 75%; Whittaker Bioproducts, 73 and 100%; Cambridge Bioscience, 89 and 100%; and University of Connecticut School of Medicine, 96 and 92%.     

Effect of antithymocyte serum on reaginic antibody formation in the rat

The effect of the time and dose of heterologous antithymocyte serum (ATS) on reaginic antibody formation was studied in the rat. Animals were treated with a single intravenous injection of ATS at various times before or after the primary immunization with dinitrophenylated Ascaris suum extract (DNP-As) and Bordetella pertussis vaccine. If animals were treated with a large lymphopenic dose of ATS shortly before or at the time of immunization, the production of reaginic as well as IgM and IgG antibodies was greatly suppressed, whereas the same treatment if given shortly after the immunization was started significantly enhanced and prolonged the reagin production. On the other hand, smaller doses of ATS even if given at the time of immunization only delayed the reaginic antibody response which also showed a marked prolongation. Furthermore, an unusual sequential production of IgM and IgG antibodies was observed in some of the ATS-treated animals. These results suggest that ATS can inhibit either inductive or regulatory function of the thymus-derived lymphocytes (T cells) depending on the time when it is administered and on the dose of ATS. Some other supporting data indicating that ATS inhibits the T cells specialized in the regulation of antibody formation are also presented.     

The influence of the ev 3 locus on the inducibility of serum antibody reactivity for envelope glycoprotein group-specific determinants

Chickens segregating for the ev 3 locus were bred by backcross matings of line 6(3) to line 15B. Analysis of RAV-1-infected segregants indicated that inducibility of antibody reactivity for envelope glycoprotein group-specific determinants correlated with the absence of ev 3, whereas noninducibility correlated with the presence of ev 3. Since the ev 3(+) and ev 3(-) segregants possessed similar genetic backgrounds, these results provide direct evidence that the ev 3 locus determines the phenotype of noninducibility.     

参地颗粒对系膜增生性肾小球肾炎大鼠CTLA-4/ B7-1介导的免疫炎症紊乱的干预作用

目的:建立系膜增生性肾小球肾炎(MsPGN)大鼠模型,观察参地颗粒对MsPGN 大鼠细胞毒性T 淋巴细胞相关 抗原-4(CTLA-4) / B7-1 介导的免疫炎症紊乱的干预作用。方法:将60 只SD 大鼠随机选取10 只作为正常组,余下50 只建立 MsPGN 大鼠模型,死亡11 只,剔除不合格3 只,共36 只大鼠成功建立MsPGN 模型,随机分为模型组、参地颗粒组和缬沙坦组,每 组12 只。参地颗粒组给予0.4 g/ (100 g·d)灌胃,缬沙坦组给予1.03 mg/ (100 g·d)灌胃,正常组和模型组给予等量温水灌胃。 12 周后观察各组大鼠24 h 尿蛋白定量和肾脏组织病理学改变,采用酶联免疫吸附(ELISA)法检测大鼠血清白介素-2(IL-2)、IL- 6、干扰素-γ(IFN-γ)、CTLA-4、B7-1 水平。结果:模型组大鼠24 h 尿蛋白定量高于正常组(P<0.05),治疗后参地颗粒组和缬沙坦 组均低于模型组(P<0.05),且参地颗粒组优于缬沙坦组(P<0.05);模型组肾组织病理示肾小球系膜细胞与系膜基质增生,治疗 后参地颗粒组和缬沙坦组均较模型组轻,参地颗粒组优于缬沙坦组(P<0.05);模型组大鼠血清IL-6、IFN-γ、B7-1 水平均高于正 常组(P<0.05),IL-2、CTLA-4 水平均低于正常组(P<0.05),治疗后参地颗粒组和缬沙坦组IL-6、IFN-γ、B7-1 水平均低于模型组 (P<0.05),IL-2、CTLA-4 水平均高于模型组(P<0.05),且参地颗粒组优于缬沙坦组(P<0.05)。结论:参地颗粒可以降低MsPGN 大鼠尿蛋白定量,减轻肾脏病理损害,其机理可能与抑制CTLA-4/ B7-1 介导的免疫炎症反应有关。     

Effect of treatment on serum antibody to Hymenolepis nana detected by enzyme-linked immunosorbent assay.

An enzyme-linked immunosorbent assay (ELISA) was developed to measure serum immunoglobulin G antibodies in 65 patients infected with Hymenolepis nana and 30 noninfected patients. Antibody was detected in 51 of 65 (sensitivity, 79%) and 5 of 30 H. nana-negative patients (specificity, 83%). Nine patients infected with H. nana were treated with praziquantel (20 to 25 mg/kg of body weight). Antibody disappeared from the sera at 90 days in six patients, five of whom had eliminated H. nana. Antibody persisted in three patients in whom H. nana infection did not clear after treatment. The H. nana ELISA had a high rate of cross-reactions with sera from patients with cysticercosis (8 of 29 [28%]) and hydatidosis (8 of 23 [35%]). The ELISA for H. nana may be useful for defining the epidemiology of H. nana infections, especially in areas free from cysticercosis and hydatidosis.     

The role of serum factors in the suppression of experimental allergic encephalomyelitis: evidence for immunoregulation by antibody to the encephalitogenic peptide.

Lewis rats immunized with myelin basic protein (MBP) in Freund's complete adjuvant (FCA) suffer from a single episode of paralysis from which they recover spontaneously. Subsequent to recovery, further episodes of paralysis cannot normally be induced by reimmunization with MBP in FCA. It is well established that serum, obtained from rats in the refractory state, can suppress the induction of experimental allergic encephalomyelitis (EAE) when given to animals from the time of immunization with MBP in FCA. Here it is shown that treatment with some such sera from Day 7 after immunization also suppressed the disease. However, not all convalescent sera were suppressive, indicating that rats immunized with MBP in FCA could become refractory to EAE without assayable levels of suppressive activity in their sera. In the context of this result it was notable that a correlation was found between the level of antibody specific for the encephalitogenic peptide in sera and the ability to suppress EAE. An inverse relationship was also shown between the amount of anti-encephalitogenic peptide antibody produced after immunization and the severity of EAE induced. Spleen cells from animals treated with Lewis anti-MBP serum after immunization with MBP in FCA could be activated to transfer EAE by in vitro culture with MBP despite the absence of any clinical signs in the donor animals, i.e. the serum inhibited the expansion or differentiation of these cells rather than preventing their priming or bringing about clonal deletion.