有寄生虫和细菌感染的三例非洲病人血清抗补体活性分析

作者对三例患有盘尾丝虫病和雅司病的非洲人进行了血清中补体情况的研究。三例病人总溶血活性及单个补体成分活性检查结果:与正常人血清比较,三例病人血清皆无溶血反应。补体成分C_1-C_4明显降低或未能测出。病人血清抗补体活性检查:三例病人血清在不同稀释浓度下,对正常豚鼠血清的溶血反应产生不同程度的抑制,若用与病人同样浓度的正常人血清,则能提高溶血活性。病人血清抑制C_4及C_2试验结果表明:病人血清抗补体活性对C_4和C_2有直接作用,对C_1或C_3无作用。并可使C_4和C_2产生温度依赖性消耗,C_4比C_2较为敏感。     

多发性骨髓瘤患者血清IL-6活性与急性相蛋白浓度的相关性研究

运用敏感的B_9细胞增殖试验检测了81例多发性骨髓瘤(MM)患者血清IL-6活性,同时分析了标本的几种急性相蛋白含量,结果表明,68％MM患者血清中IL-6活性大于5μ/ml(正常对照为5μ/ml以下),几种急性相蛋白中C-反应性蛋白(CRP)在MM时升高(P<0.01),平均达正常对照组的17倍以上,MM患者补体C_4与正常对照组无差异(p>0.05),C_3、白蛋白及转铁蛋白在MM时分别比正常下降24.42％、38.83％和32.80％,且与疾病分期有关,在血清IL-6大于5μ/ml的55例中,IL-6活性与CRP、C_3、白蛋白的相关系数分别为0.46,-0.34和-0.29,IL-6与转铁蛋白浓度相关不明显。本文结果提示:CRP、C_3及白蛋白等含量的变化可作为反映MM病情的简易而敏感的指标。     

新生儿肺出血时血清补体的变化

本文对21例正常新生儿脐血血清和10例肺出血患婴血清C_(1q)、C_4和C_3用单向扩散法作定量分析;对14例正常新生儿脐血血清和全部肺出血患婴血清用交叉免疫电泳法检测C_3裂解产物。21例正常新生儿脐血血清C_(11)、C_4 和 C_3含量(x±SD)分别为 13.3±4.0、17.4±6.3和81.1±25.2mg％。10例肺出血患婴分别为 5.2±2.4、11.7±6.8和 51.2±28.8mg％。两组相比均有明显差异(P<0.001、0.01和 0.01)。14例正常新生儿脐血EDTA血清全部未能检出C_3裂解产物,10例肺出血患婴中 4例检出 C_3裂解产物。表明肺出血患婴存在明显的低补体血症和补体激活。结合10例肺出血患婴中8例存在感染,表明严重感染的肺出血患婴伴有的低补体血症,可能与细菌抗原与相应抗体形成的免疫复合物或细菌抗原直接激活补体,导致补体消耗增加有关。     

测定C_4活性的改良溶血试验

C_4是补体经典激活途径的一个重要组分,它的变化对一些自身免疫性疾患的研究有着重要的意义。本文介绍了一种测定血清中C_4活性的改良溶血试验,方法简便,试剂用量少,具有特异性,去除了C_3干扰。本文还摸索了测定中各种影响因素,采用了质控措施。初步应用于临床,56例正常人血清C_4溶血活性为8270单位/毫升,而急性肾炎,系统性红斑狼疮等疾患的患者,其C_4溶血活性均低于正常人组。     

家兔高胆固醇性动脉硬化时血清C_3的变化及C_3、IgG在粥样硬化斑中的沉积

我们曾报道,静脉内注射异种蛋白质可以加重高胆固醇性动脉硬化的程度,并提出这种作用可能和免疫复合物在动脉壁上的沉着所引起的血管壁损伤有关。一般认为,这种血管壁的损伤主要是补体系统被激活而导致的。为探讨这一现象的机制,我们对家兔实验性动脉硬化时血清的总补体,补体第三成分C_3、血清的抗补体活性进行了测定,并用免疫荧光法对动脉粥样硬化斑块中的C_3和IgG做了检查,结果表明,无论是单独喂胆固醇饲料或者合并注射异种蛋白质的家兔,血清C_3都明显降低,并都可在粥样硬化斑块中查出IgG和C_3。这一结果进一步提示了动脉硬化的发病与机体疫反免应的密切而复杂的关系。     

激活补体类HBsAg循环免疫复合物与乙型肝炎肝损害关系初探

对六类236例乙型肝炎血清标本并行检测激活补体类HBsAg免疫复合物、GPT和C_3。在SGPT升高组和血清C_3降低组中,激活补体类HBsAg循环免疫复合物的检出率分别非常显著地高于SGPT和血清C_3正常组。但进一步型内分析发现,在重型乙型肝炎及慢性活性乙型肝炎者中,激活补体类HBsAg循环免疫复合物的检出率与SGPT升高或血清C_3降低有关;而在HBsAg无症状携带者、急性乙型肝炎、慢性迁延性乙型肝炎及乙型肝炎后肝硬化者中,其检出率与之无关。这些结果提示,与HBsAg循环免疫复合物有关的肝损伤作用,在急性乙型肝炎和慢性迁延性乙型肝炎中不起主要作用,而在重型乙型肝炎和慢性活动性乙型肝炎中则起重要作用;激活补体类HBsAg循环免疫复合物还是导致乙型肝炎患者补体激活和消耗的重要原因之一。     

C_3肾炎因子(C_3NeF)交叉免疫电泳测定法

<正> 1969年 Spitzer 等发现膜增殖性肾小球低补体血症(membranoproliferative glo-merulonephritis,简称 MPGN)病人血清中存在一种能使 C_3裂解的活性物质,命名为 C_3肾炎因子(C_3 Nephritic factor 简称 C_3NeF)。Koch 等发现 C_3NeF 与正常人血清中的辅助因子在 Mg~(++)存在下能形成一活性物质,并称之为 C_a 溶解肾炎因子(C_3 Lytic     

CA15-3IRMA对乳腺癌诊断价值的探讨

应用灵敏、特异的免疫放射分析(IRMA)对60例正常人、44例乳腺癌患者(手术前、后)93例其它恶性肿瘤患者及147例良性病变患者血清CA15-3进行检测,正常值为17.9±5.7u/ml,若以(?)+2SD为阳性界值,则CA15-3为30u/ml。44例乳腺癌术前CA15-3平均水平为62.4±57.3u/ml,>30u/ml的有29例,占65.9％;34例行乳癌根治术后CA15-3为17.8±8.4u/ml,>30μ/ml的仅4例;45例乳腺良性病变患者CA15-3为15.3±7.9u/ml,仅2例阳性,假阳性率为4.4％。乳腺癌组CA15-3水平明显高于乳腺良性病变组和正常组(p均<0.001)。肝癌、胃癌、结-直肠癌患者血清CA15-3均不同程度地升高,但不如乳腺癌患者明显,其值分别为39.5±32.3u/ml、31,6±35.2u/ml、24.2±25.8u/ml;CA15-3>30u/ml的阳性率分别为42.0％、24.2％、20.6％。102例良性病变患者血清CA15-3为17.2±6.6u/ml,>30u/m1的有10例,总的假阳性率为9.8％。结果表明,从灵敏度与特异性两方面总体考虑,CA15-3是乳腺癌诊断的良好标志物。     

肿瘤标志物CA—242IRMA的临床应用

CA-242是继CA-50之后出现的较新的肿瘤标志物之一。近年来,我们将这一新项目运用于临床,获得了一些收获,本文将结果加以总结,并对其在肿瘤诊断中的应用价值作一初步评价。 材料和方法 一、材料和试剂:CA-242IRMA试剂盒由中国科学院肿瘤研究所提供。正常人血清均为本院体检合格者,各类良恶性肿瘤患者血清取自本院就诊者,静脉取血,分离血清后-20℃保存待用,诊断皆经病理证实。 二、CA-242测试:采用一步法进行,于标准管和样品管中分别加入50μl标准CA-242抗原和待测血清,加入100μl保温液及100μl~(125)I-CA-242,再加入单抗包被珠一粒,置4℃过夜。吸去上清,用蒸馏水洗珠三次,吸干后测珠上的放射性活度,从标准曲线中读取CA-242的含量。 结果和讨论 68例正常人血清CA-242的水平为2.3±7.2u/ml,范围0～9u/ml,如以(?)±2SD计算,阳性界值为17u/ml,此时正常人的假阳性率3.4％。103例良性疾病血清CA-242值为3.0±6.8u/ml,与正常人之间无显著差别。108例恶性肿瘤患者血清CA-242值为49.6±89.1u/ml,与正常组以及良性病组相比较有显著差别。恶性肿瘤CA-242总的阳性率为67.4％,其中胰腺、胆囊恶性肿瘤的阳性率达92.2％,而假阳性率较低为13.2％(表1)。     

一种测定旁路激活途径中补体活性的方法

补体激活的旁路途径中补体活性的测定方法至今没有统一的标准,现有的方法都是根据非致敏的家兔红细胞具有激活旁路途径的特性而设计的,观察指标为相对吸光度,难以准标化。作者以兔红细胞50％溶血为标准,定量测定补体旁路激活途径的补体活性,并对其在类风湿性关节炎诊断中的意义进行了研究。实验过程:在4支试管中分别注入0.02ml、0.04ml、0.06ml、0.08ml人血清,     

Generation of complement anaphylatoxins and C5b-9 by crystalline cholesterol oxidation derivatives depends on hydroxyl group number and position

Cholesterol crystals activate the human alternative complement pathway. Loss of Factor B hemolytic activity in C2-deficient serum was comparable to that in normal human serum after incubation with cholesterol crystals. Consumption of Factor B hemolytic activity in normal serum incubated with cholesterol occurred in a time- and dose-dependent manner. The reduced capacity of crystals-absorbed serum to activate C2, but not Factor B, on fresh crystals, indicated that cholesterol mediates antibody-dependent classical pathway activation in addition to alternative pathway activation in whole serum. Cholestane triol, an oxidation derivative of cholesterol which bears three hydroxyl groups, cleaved 5-fold more C3 than cholesterol in normal human serum. Three cholesterol derivatives, each bearing two hydroxyl groups, were intermediate activators between cholesterol and cholestane triol. The compounds differed, however, in their complement-activating ability, indicating that hydroxyl position as well as number exerts an influence on complement activation. Measurements of C3adesArg and C5adesArg antigens in cholesterol crystal treated serum revealed that approx. 10% of total serum C3 was cleaved and that, on a molar basis, only 3% C5 cleavage occurred relative to C3 cleavage. For 1 mole of C5a generated, 0.1 moles of fluid-phase C5b-9 was detected. Although the extent of C3 cleavage varied with each cholesterol derivative depending on the position and number of hydroxyl groups, the relative coupling efficiency of C3 and C5 cleavage and C5a and C5b-9 generation was similar for all compounds. The ability of cholesterol and its oxidation products to generate anaphylatoxins and C5b-9 complexes may be of importance in mediating inflammatory processes involved in atherogenesis.     

Immunological properties of glycolipids from membranes of Acholeplasma laidlawii.

Glycolipids, the predominant class of lipids in the membranes of Acholeplasma laidlawii, are the haptenic determinants that react with anti-A. Laidlawii serum to fix complement. The predominant complement-fixing activity of the membrane glycolipids was associated with the monoglucoysyl diglyceride, diglucosyl diglyceride, glycerlphosphoryl diglucosyl diglyceride (GPDD), and an unknown lipid B, which did not react with ninhydrin but release glucose and glycerol and traces of phosphorus upon hydrolysis. The glycolipids monoglucosyl diglyceride and diglucosyl diglyceride or GPDD and unknown lipid B were paired as a result of their cross-reactions with selective antisera prepared with the aid of reconstituted membrane complexes containing membrane lipids. Reconstituted membrane complexes assembled from [14C]monoglucosyl diglyceride and delipidated membrane proteins gave optimal complement fixation titers before saturation of the complexes with the ]14C]monoglucosyl diglyceride. The phosphoglycolipid of the membrane, GPDD, was anticomplementary as a pure lipid, a cholesterol liposome, and a reconstituted membrane complex. This anticomplementary activity, which was caused by 3 mug of pure GPDD, affected both human and guinea pig complement. Although human C1, C4, C3, and C5 were not inhibited by GPDD, C2 was inhibited 10-fold by reconstituted membrane complexes containing 150 mug of GPDD. A role for this phosphoglycolipid is discussed in the hypothetical mechanism of inhibition of C2 attachment to SAC1, 4 sites.     

Activation of complement system by porins extracted from Salmonella typhimurium

The effect of porins purified from Salmonella typhimurium on the complement system was investigated both in vitro and in vivo. Incubation of porins with either human or guinea pig serum resulted in the consumption of the total complement activity when an amount of porins ranging from 8 to 10 micrograms per 100 microliters of serum was used. The activation of the complement system was temperature dependent, suggesting an active process rather than passive adsorption of the complement components by porins. In addition, the activation had a fast kinetic and proceeded mainly through the classical pathway. This conclusion is supported by the consumption of C1s and C4 in normal human serum treated with porins and also by the depletion of C3 activity in the C1s-deficient serum which was marked only when purified C1s was added to the serum before incubation with porins. Injection of 100 micrograms of porins into guinea pigs induced profound complement consumption at 6 h postinjection that persisted up to 12 h. We conclude from this study that porins can effectively contribute to complement activation and to subsequent biological events induced by gram-negative bacteria.     

Activation of human complement by rat peritoneal mast cells and its inhibition by a rat serum factor

The effects of normal sera from humans, rats, and guinea pigs on unsensitized rat peritoneal mast cells were studiedin vitro. Five to 20% fresh human sera induced mast cell death and substantial histamine release. The factor was heat labile. Neither hereditary C3-deficient sera nor experimentally Clq-depleted sera showed cytotoxicity. The CH50 activity of human serum was decreased to about one half after a 15-min incubation with 2×10⁶ mast cells/ml at 37°C. The cytotoxic activity and CH50 reduction were completely eliminated by an addition of 10 mM Mg-EGTA to the serum. These data demonstrated that unsensitized rat mast cells served as both the initiator and target of complement activity when human serum was used as a complement source. Requirements of both Ca⁺⁺ and Clq suggested the activation of the classical pathway of complement. Fresh 5–20% sera from rats and guinea pigs, on the other hand, showed neither cytotoxicity nor CH50 reduction. Furthermore, these sera strongly inhibited the human serum-induced reaction. The latter results indicated the presence of a modulating factor in rat and guinea pig sera, which inhibits mast cell associated complement activation.     

BrkA protein of Bordetella pertussis inhibits the classical pathway of complement after C1 deposition

Bordetella pertussis produces a 73-kDa protein, BrkA (Bordetella resistance to killing), which inhibits the bactericidal activity of complement. In this study we characterized the step in the complement cascade where BrkA acts, using three strains: a wild-type strain, a strain containing an insertional disruption of brkA, and a strain containing two copies of the brkA locus. Following incubation with 10% human serum, killing was greatest for the BrkA mutant, followed by that for the wild-type strain, while the strain with two copies of brkA was the most resistant. Complement activation was monitored by enzyme-linked immunosorbent assay (ELISA) or Western blotting. ELISAs for SC5b-9, the soluble membrane attack complex, showed that production of SC5b-9 was greatest with the brkA mutant, less with the wild type, and least with the strain containing two copies of brkA. Deposition of complement proteins on the bacteria was monitored by Western blotting. A decrease in deposition on the bacteria of C4, C3, and C9 corresponded with decreased complement sensitivity. Deposition of C1, however, was not affected by the presence of BrkA. These studies show that BrkA inhibits the classical pathway of complement activation and prevents accumulation of deposited C4.     

Dichotomy between opsonization and serum complement activation by encapsulated staphylococci.

Previous studies have demonstrated that encapsulated Staphylococcus aureus strains are not effectively opsonized by the serum complement system. Encapsulated staphylococci thereby "resist phagocytosis." To test whether this phenomenon might be explained by an inability of encapsulated strains to activate complement, the relationship between staphylococcal opsonization and serum complement activation was studied. Although encapsulation was found to interfere with opsonization by pooled human serum (human polymorphonuclear leukocytes phagocytized significantly fewer encapsulated bacteria than unencapsulated bacteria after incubation in this opsonic source), encapsulated (S. aureus M and Smith diffuse) and unencapsulated (S. aureus M variant and Smith compact) strains had similar capacities for complement activation as measured by C3-C9 consumption. When C2-deficient and immunoglobulin-deficient sera were studied, again C3-C9 consumption was not influenced by the presence or absence of a capsule. In addition, C3 was detected on the surface of both S. aureus M and M variant strains after incubation in pooled serum and staining with fluorescein-conjugated anti-C3 antibody. Thus, encapsulated staphylococci are not effectively opsonized even though complement is activated and C3 is present on the bacterial surface. The exact mechanism by which the capsule interferes with opsonization is still not known; however, inhibition of complement activation appears not to be the explanation of this phenomenon.     

Deposition of complement activation products on plastic-adsorbed immunoglobulins. A simple ELISA technique for the detection of defined complement deficiencies

The activation of complement components in human serum has been studied using immunoglobulins adsorbed to microtiter plates. The sequential deposition of complement fragments was detected by a series of mono- and polyclonal antibodies in an indirect enzyme-linked immunosorbent assay (ELISA). Antibodies against C1q, C1s, C4b/d, C3b/d, factor B, C5b-9 membrane attack complex (MAC), the regulatory complement proteins C4 binding protein (C4bp) and properdin were reactive. Several lines of evidence suggest that complement activation was via the classical pathway: (1) complement activation was highly isotype-restricted with regard to the adsorbed Igs (human IgG1 and IgG3 as well as mouse IgM, IgG2a and IgG2b isotypes are strong activators in contrast to human IgG2, IgG4, IgA and mouse IgG1); (2) Ca2+ depletion, heat treatment (56 degrees C for 45 min), incubation with 0.5 M KSCN or heat-aggregated immunoglobulins (aggIgG) abrogated serum activity; (3) complement deficient sera (C1q def', C2 def', C6 def' human sera; C2 def', C4 def' guinea pig sera) showed impaired deposition of the complement components that follow the missing component in the cascade of activation. In a clinical study sera from patients with systemic lupus erythematosus (SLE) were investigated in order to measure the effect of hypocomplementemia due to complement consumption. The results obtained suggest that this new and simple assay is well suited for (1) the detection of various inherited complement deficiencies, (2) the semiquantitative evaluation of sera with decreased complement levels, (3) a more detailed study of complement components bound to a solid phase.     

Activation of Classical Complement Pathway by Naturally Occurring Heteroantibodies in Normal Rabbit Serum. Effect on Subpopulations of Human Lymphoid Cells

Heteroantibodies present in normal rabbit serum (NRS) are toxic to human B lymphocytes, T lymphocytes, and monocytes. Even NRS, which exhibits little back ground cytotoxicity for human lymphoid cells in conventional HLA or B-cell lymphocytotoxic assays, can be shown to contain considerable activity by making two modifications in usual procedures: by washing cells in saline or balanced salt solutions devoid of protein or sugar substances, and by increasing incubation time for 1 h to 3--4 h. Using such modifications, the cytotoxic activity of NRS towards human lymphoid cells was investigated and was found to involve activation of the classical complement pathway rather than activation of the alternate complement pathway. Residual unwanted background cytotoxicity of NRS toward human lymphoid cells can be decreased without loss of desired complement activity either by heating NRS for 15 min at 50 degrees C or by mixing NRS with small amounts of normal human serum.     

Activation of the alternative pathway of complement by monosodium urate crystals

Monosodium urate crystals (MSU) have been shown to activate the alternative pathway of complement in a dose- and time-dependent fashion at 37 degrees C. Activation was maximal upon addition of 10-20 mg/ml monosodium urate crystals to C2-deficient human serum (C2D) or normal human serum containing 5 mM MgEGTA. Immunoelectrophoretic analysis of such treated sera demonstrated cleavage of C3 and factor B. Incubation of highly purified C3 and factor B with 10 mg/ml MSU did not, however, affect their immunoelectrophoretic pattern, suggesting that cleavage of either factor B or C3 in serum requires an intact alternative complement pathway. The fluid-phase control proteins, Factor H and Factor I, were not found to be diminished upon incubation of C2D serum or NHS containing MgEGTA with MSU. Thus activation appeared to be surface dependent and not a consequence of control protein depletion. It was also found, in agreement with earlier observations, that the classical complement pathway is activated, with concomitant depletion of C1 and C4. We conclude that MSU crystals activate both the classical and alternative pathways, and that such activation may participate in the pathogenesis of gouty arthritis.     

Characterization of alternative pathway inhibition by a serum derived, low molecular weight complement inhibitor.

Normal human serum contains an inhibitor of complement which is distinguished by its small size of 500 daltons, the low molecular weight inhibitor (LMWI). When LMWI was present during incubation of zymosan or cobra venom factor with serum, formation of complement reactive complexes was blocked as measured by failure of these mixtures to lyse susceptible erythrocytes from patients with paroxysmal nocturnal haemoglobinuria (PNH). Addition of LMWI to pre-formed complexes had no effect on their subsequent haemolytic activity. When dialysis was used to remove LMWI from reaction mixtures, it was shown that LMWI had not irreversibly altered any of the complement components. Purified components were used to demonstrate that LMWI prevented factor D activation of cobra venom factor-factor B complexes. LMWI also strongly inhibited binding of 125I-factor B to human erythrocytes bearing C3b and had little or nor effect on binding of 125I-factor H to the C3b bearing cells. Factor B binding to C3b was equally inhibited on normal and PNH erythrocytes. Thus, a dialysable fraction from normal human serum prevents activation of human complement by blocking formation, but not the activity of the C3/C5 convertase. These low molecular weight inhibitors result in inhibition of factor B binding to C3b and inhibition of factor D activation of C3bB complexes.