恒河猴红细胞免疫功能的研究

本文应用花环法测定了健康恒河猴红细胞上C_(3b)受体和免疫复合物的数量。恒河猴红细胞上C_(3b)受体花环率的数量为12.83±1.95％,免疫复合物花环率为6.37±1.25％。研究结果表明恒河猴的红细胞,除了具有携氧、运输气体等功能外,还具有重要的免疫功能。     

温度对中华大蟾蜍和中华鳖冬眠前离体红细胞免疫功能及其大小的影响

为了探讨温度变化对中华大蟾蜍(Bufo gargarizans)和中华鳖(Trionyx sinensis)离体红细胞免疫功能及细胞大小的影响,我们设计了10℃、20℃、30℃、37℃ 4个温度组,通过红细胞C3b受体花环试验和红细胞免疫复合物花环试验检测红细胞免疫活性,同时测量其红细胞大小.结果表明,温度对中华大蟾蜍离体红细胞C3b受体花环率影响显著,随着温度的升高,中华大蟾蜍离体红细胞C3b受体花环率逐渐降低,在30℃和37℃时,其红细胞C3b受体花环率明显比10℃时低;而温度对其离体红细胞免疫复合物花环率和红细胞大小却没有显著影响.温度对中华鳖离体红细胞C3b受体花环率、免疫复合物花环率及其大小亦没有显著影响.除了10℃时中华鳖离体红细胞免疫复合物花环率与中华大蟾蜍没有明显差别外,其余各温度下中华鳖离体红细胞C3b受体花环率和免疫复合物花环率均明显比中华大蟾蜍的高,而其红细胞的长径和短径却明显比中华大蟾蜍红细胞的小.这说明,温度能明显影响中华大蟾蜍离体红细胞免疫功能,而对中华鳖离体红细胞的免疫活性影响不明显,随着动物进化程度的提高,中华鳖红细胞对温度变化的适应能力和免疫活性比中华大蟾蜍明显增强.     

分枝杆菌L型感染与肺癌患者红细胞免疫功能改变的相关性

目的:研究分枝杆菌L型血行感染与肺癌患者红细胞免疫功能改变的相关性.方法:溶血离心培养法和滴片法检测血液中的分枝杆菌L型,常规方法测定红细胞沉降率(ESR)、红细胞C3b受体花环率(C3bRR)、免疫复合物花环率(ICRR)、血清红细胞C3b受体花环促进率(RFER)和抑制率(RFIR).结果:TBL( )与TBL(-)肺癌患者相比,C3bRR和ICRR两项指标均明显降低.TBL( )肺癌与TBL(-)肺癌患者相比,ESR增快更为显著.TBL(-)较TBL( )之RFER值的降低更为明显.RFIR差异未见有显著性.结论:分枝杆菌L型血行感染影响肺癌患者血沉和红细胞免疫功能改变.     

中医药红细胞免疫强化作用的研究进展

近 10年来对红细胞免疫功能研究 ,证实红细胞免疫功能有清除免疫复合物 ,促进吞噬作用 ,识别抗原 ,提呈抗原 ,免疫调节。郭峰建立了简易的红细胞C3 b受体花环试验与红细胞免疫复合物花环试验 ,分别测定红细胞膜C3 b受体活性和红细胞膜上吸附免疫复合物的情况。红细胞的生成、发育及功能状态与中医脾、肾密切相关。中医“正邪斗争”就是红细胞免疫 ;中医辩证 ,就是红细胞识别抗原 ,提呈抗原 ;中药施治 ,就是红细胞清除循环免疫复合物 ,抗感染、抗病毒。中医药在肿瘤、自身免疫性疾病 (类风湿性关节炎、肾病综合症、I型糖尿病、牛皮癣 )艾滋病和肺气肿急性感染期等病治疗中 ,采用花环试验测定 ,结果显示RBC C3 bRR明显提高 ,RBC ICRR显著降低。能增强细胞免疫 ,促进淋巴细胞转化及E 玫瑰花环形成的中药很多 ,为提高红细胞免疫功能展现了广阔前景。至于血粘度和红细胞免疫的关系还有待进一步探讨。     

鸡红细胞免疫吸附性能与白细胞数量关系

应用红细胞C3b受体花环(G3b Receptor Rosette,G3bRR)和复合物花环(Immu Complex Rosette,ICR)试验证实鸡红细胞表面存在补体C3b受体(C3bR),表明红细胞免疫系统RCIS(Red Cell Immune System)的概念适用于这种动物;并且通过对鸡的白细胞的计数,证明了红细胞免疫吸附性能与白细胞数量之间存在一定的正相关性。     

人血清补体3b的提纯与鉴定

据报道红细胞膜上有补体3b(C_(3b))的受体。阵发性睡眠性血红蛋白尿症(PNH)是属红细胞的膜病变,对补体较为敏感,易产生溶血,可能是因为膜上C_(3b)受体与正常红细胞不同。为了研究C_(3b),与膜上C_(3b)受体结合情况,并进一步分离膜上C_(3b)受体,我们制备了纯的C_(3b)。一、材料与方法 1.C_3的提纯见文献[2] 2.免疫双扩散见文献[3] 3.C_3的垂直板型聚丙烯酰胺凝胶电泳(PAGE)见献文[2] 4.C_3分子量测定按Pharmacia标准蛋白测分子量法     

健康人群及某些疾病红细胞C_3d受体活性的观察

近年来,红细胞对机体免疫功能的作用日益受到人们的重视。证明红细胞表面的C_3b受体能与循环系统中的抗原—抗体—补体复合物粘附在一起。这种免疫粘附作用(RCIA)不但可能使循环复合物得到清除。而且T细胞区亦得以依赖反应等。     

高住低练对游泳运动员红细胞免疫黏附功能及白细胞计数的影响

目的:了解高住低练对高水平游泳运动员血细胞天然免疫功能的影响。方法:6名女子游泳运动员进行2周高住低练,训练前、训练期间及恢复1周分别测试红细胞黏附功能、白细胞计数指标。结果:训练期间运动员红细胞C3b受体花环率(RBC-C3bRR)明显下降(P<0.05),红细胞免疫复合物花环率(RBC-ICR)明显上升(P<0.05),恢复1周后指标回归正常;白细胞、粒细胞在训练期总体上呈显著性下降(P<0.05),训练结束1周后恢复;单核细胞和淋巴细胞训练期间总体趋势是下降,训练结束1周后仍未恢复。结论:红细胞和粒细胞的天然免疫功能下降快,恢复快,但淋巴细胞、单核细胞计数恢复较慢,运动员对本次训练基本适应。     

S.D和Wistar大鼠红细胞免疫功能测定

生命科学中药理、毒理和免疫学实验常用的大鼠有Ｓ．Ｄ和Ｗｉｓｔａｒ两个品种。其白细胞系统免疫功能状态已有报导,但关于红细胞免疫功能研究甚少。我们以红细胞Ｃ３ｂ受体酵母花环率和红细胞免疫复合物花环率为指标,检测了该两个品种大鼠的红细胞Ｃ３ｂ受体免疫粘附功...     

高原鼠兔红细胞免疫功能的研究

本文应用免疫花环试验法测定了封闭群中高原鼠兔红细胞Ｃ３ｂ受体和红细胞免疫复合物的花环率,并比较了不同年龄高原鼠兔红细胞的免疫功能。研究结果表明,高原鼠兔红细胞在自身免疫系统中具有重要的免疫功能;随年龄增长,其红细胞免疫功能逐渐减弱。与人、兔和大白鼠等动物相比,高原鼠兔红细胞免疫粘附活性高,因此适宜作红细胞免疫实验的动物模型。     

Characterization of the C3 receptor induced by herpes simplex virus type 1 infection of human epidermal, endothelial, and A431 cells

Herpes simplex virus type 1 (HSV-1) infection induces the appearance of viral analogues of human Fc IgG and C3 receptors on the surface of human cells. The virally induced C3 receptor(s) has been broadly defined as a C3b receptor, but its ligand binding characteristics have not been rigorously defined. In this study, human epidermal cells, A431 cells, and human umbilical vein endothelial cells infected with HSV-1 demonstrated rosetting with sheep erythrocytes (E) coated with IgG (E-IgG) or the complement components C3b (EAC3b) or iC3b (EAC3bi), but not with E-IgM, C4 (EAC14), C3d (EAC3d), or E alone. Rosetting was markedly enhanced by pretreatment of HSV-1-infected cells with neuraminidase. Unlike human C3 receptors, the HSV-1-induced C3 receptor was found to be trypsin resistant. To determine whether HSV-1 induced CR1-like receptors or CR3-like receptors, infected cells were pretreated with EDTA, which is known to inhibit native CR3 function. EDTA failed to prevent rosetting with EAC3bi. Furthermore, blocking studies using monoclonal antibodies against CR1 and CR3 revealed that the anti-CR1 antibody 5C11 consistently blocked EAC3b and EAC3bi rosetting with HSV-1-infected cells in a dose dependent manner, but monoclonal antibodies against CR3 did not. This study indicates that the HSV-1-induced C3 receptor is an analogue of CR1.     

Analysis of C3-receptor activity on human B-lymphocytes and isolation of the complement receptor type 2 (CR2).

The rosetting of defined C3-fragment-coated sheep erythrocytes to B-cell-enriched tonsil lymphocytes was measured. The rosetting lymphocytes were homogeneous with respect to expression of C3b, iC3b and C3d receptors. Isolation of receptors for C3 fragments from surface-radioiodinated lymphocytes by affinity chromatography on immobilized C3u, iC3b and C3d,g produced two proteins with partially overlapping specificities. A protein of 240 000 Mr, recognized by the monoclonal antibody To5 and identified as CR1 (complement receptor type 1), had affinity for C3u and iC3b. A protein of 145 000 Mr, recognized by the monoclonal antibody B2, had affinity for all three C3 fragments. Inhibition of rosetting by antibodies to these proteins indicates that CR1 is responsible for C3b-mediated rosetting and that the 145000-Mr receptor (CR2) is responsible for C3d-mediated rosetting. Partial inhibition by both anti-CR1 and anti-CR2 antibodies of iC3b-mediated rosetting indicates that both receptors are involved in iC3b-mediated rosetting. No other protein appears to be involved in tonsil B-cell rosetting to C3-fragment-coated cells. A method for preparing CR2 from tonsil lymphocytes based on affinity chromatography on C3d,g-Sepharose has been developed. Forty tonsil pairs (2 X 10(10) B-cells) yield about 40 micrograms of pure protein equivalent to a purification of 6500-fold from a detergent extract.     

Inhibition of complement-mediated functions of human neutrophils by impermeant stilbene disulfonic acids

The impermeant labeling reagents 4,4'-diisothiocyanostilbene-2-2'-disulfonic acid (DIDS) and 4-acetamido-4'-isothiocyano-2,2'-disulfonic acid (SITS) inhibited in a concentration-related manner the enhanced generation of superoxide radicals (O2) by human neutrophils engaged in the phagocytosis of zymosan that had been opsonized in fresh serum, without altering the O2 generation by neutrophils exposed to zymosan opsonized in heat-decomplemented serum or to phorbol myristate acetate (PMA). That the stimulus specificity of the suppression of O2 generation by SITS and DIDS is predominantly attributable to an action on neutrophil plasma membrane receptors for complement was suggested by the similarity of the concentration dependence of the inhibition of the expression of neutrophil C3b receptors, as assessed by a rosetting assay. Washing neutrophils that had been pretreated with the covalent label DIDS failed to reverse either the suppression of C3b-dependent rosetting or the inhibition of O2 generation stimulated by opsonized zymosan. In contrast, pretreatment with DIDS and washing or erythrocytes bearing C3b and of opsonized zymosan did not inhibit their capacity to form rosettes and to stimulate O2 generation by neutrophils, respectively. In the same rosetting assay, the expression of IgG-Fc receptors was unaffected by SITS and DIDS. The rapid and apparently selective inhibition of the expression of neutrophil C3b receptors by noncytotoxic concentrations of the impermeant stilbene disulfonic acids may provide a means to analyze the complement dependence of other neutrophil effector functions.     

C3 modified at the thiolester site: Acquisition of reactivity with cellular C3b receptors

Isolated human C3 protein has been reported to have variable affinity for cellular C3b receptors. The question was investigated therefore as to whether native C3 or a derivative form of the protein exhibits receptor affinity. It was found that native C3 lacks the ability to react with C3b receptors. However, C3 modified at the thiolester site, either by treatment with chaotropic agents or methylamine or by freezing and thawing, expressed inhibitory activity in the C3b-mediated rosetting assay or immune adherence. The extent of inhibition was comparable to that caused by C3b. The ability of modified C3 to react with cellular C3b receptors constitutes an additional functional property of C3b-like C3.     

Mapping of the region of complement receptor (CR) 1 required for Plasmodium falciparum rosetting and demonstration of the importance of CR1 in rosetting in field isolates

The malaria parasite Plasmodium falciparum induces a number of novel adhesion properties in the erythrocytes that it infects. One of these properties, the ability of infected erythrocytes to bind uninfected erythrocytes to form rosettes, is associated with severe malaria and may play a direct role in the pathogenesis of disease. Previous work has shown that erythrocytes deficient in complement receptor (CR) 1 (CR1, CD35; C3b/C4b receptor) have greatly reduced rosetting capacity, indicating an essential role for CR1 in rosette formation. Using deletion mutants and mAbs, we have localized the region of CR1 required for the formation of P. falciparum rosettes to the area of long homologous repeat regions B and C that also acts as the binding site for the activated complement component C3b. This result raises the possibility that C3b could be an intermediary in rosetting, bridging between the infected erythrocyte and CR1. We were able to exclude this hypothesis, however, as parasites grown in C3-deficient human serum formed rosettes normally. We have also shown in this report that rosettes can be reversed by mAb J3B11 that recognizes the C3b binding site of CR1. This rosette-reversing activity was demonstrated in a range of laboratory-adapted parasite strains and field isolates from Kenya and Malawi. Thus, we have mapped the region of CR1 required for rosetting and demonstrated that the CR1-dependent rosetting mechanism occurs commonly in P. falciparum isolates, and could therefore be a potential target for future therapeutic interventions to treat severe malaria.     

An immobile subset of plasma membrane CD11b/CD18 (Mac-1) is involved in phagocytosis of targets recognized by multiple receptors

CD11b/CD18 is a heterodimeric leukocyte surface receptor which functions in both C3bi-ligand binding and homotypic and heterotypic cell adherence. We have examined the effect of several anti-CD11b/18 mAb on phagocytosis of IgG (EIgG) or complement (EC4b) opsonized erythrocytes by polymorphonuclear leukocytes (PMN) and monocytes. F(ab')2 of two mAb (IB4, an anti-beta-chain mAb and Mo-1 an anti-alpha-chain mAb), inhibited both phagocytosis of EIgG and phorbol ester-stimulated phagocytosis of EC4b by PMN and monocytes. These F(ab')2 inhibited the binding of EIgG to monocytes, but they had no effect on binding of EIgG to PMN, or EC4b to either phagocyte. In addition, IB4 inhibited phorbol-ester stimulated phagocytosis of sheep E opsonized with C component 3bi (EC3bi) without inhibiting rosetting of these same targets. These data separate the anti-phagocytic effect of these mAb from effects on phagocyte-target adherence. When PMN were adherent to an anti-CD11b/CD18 F(ab')2-coated surface, EC3bi binding was abolished, but phagocytosis of EIgG or EC4b was unaffected. Subsequent addition of fluid- phase IB4 or Mo-1 F(ab')2 inhibited phagocytosis of EIgG or EC4b by the adherent cells. This suggested that the CD11b/CD18 involved in C3bi rosetting were mobile in the membrane, whereas those involved in phagocytosis of EIgG or EC4b were not. Cytochalasin treatment of PMN during adherence to F(ab')2-coated plates decreased both apical expression of CD11b/18 and subsequent ingestion of EIgG by 70%, suggesting that microfilaments are important in maintaining immobile CD11b/18 on the apical PMN surface. We conclude that there are functionally distinct populations of CD11b/CD18 on monocytes and PMN: one involved in C3bi rosetting and another involved in the process of phagocytosis mediated via several different receptors. CD11b/18 is not required for optimal target binding in all cases, but is always required for ingestion. As with several other integrins, the CD11b/18 molecules involved in phagocytosis have a functional association with the cell cytoskeleton.     

Human C5a modulates monocyte Fc and C3 receptor expression

FcIgG and C3 (CR1 and CR3) receptors are responsible for binding opsonized particles, phagocytosis, and immune adherence reactions by circulating and tissue-fixed mononuclear phagocytes. Alterations in the expression of these receptors may thus significantly influence the function of these cells. Because chemoattractants have been shown to both recruit and modulate the function of monocytes, this study specifically examines the effects of human C5a and N-formyl-methionyl-leucyl-phenyl-alanine (FMLP) on human peripheral blood monocyte FcIgG and C3 receptor expression in vitro. Adherent, elutriator-purified monocytes were incubated with C5a (10(-7) to 10(-10) M) or FMLP (10(-5) to 10(-10) M) for 30 min at 37 degrees C, and FcIgG receptor expression was assessed by rosetting with sheep erythrocytes sensitized with limiting dilutions of IgG. Human C5a caused dose-related increases in Fc rosettes of 28% at 10(-9) M, 63% at 10(-8) M, and 167% at 10(-7) M (p less than 0.01). In contrast, no significant increases in monocyte Fc receptor expression were induced by FMLP. Similar rosetting experiments were performed with sheep erythrocytes opsonized with limiting amounts of human C3b to assess C3b receptor expression on adherent human monocytes stimulated with C5a (10(-7) to 10(-10) M) or FMLP (10(-6) to 10(-9) M) for 30 min at 37 degrees C. Again, human C5a caused dose-related increases in monocyte C3b rosette formation; at 10(-8) M and 10(-7) M concentrations of C5a, these increases equaled 119% and 196%, respectively (p less than 0.05). In these experiments, 10(-6) M FMLP also caused a significant increase of 110% in monocyte C3b rosette formation (p less than 0.05). Modulation of monocyte cell surface receptors by human C5a or FMLP was also examined by measuring cell fluorescence and side scatter by dual channel flow cytometry after staining normal leukocytes in citrated venous blood with receptor-specific monoclonal antibodies. These flow cytometric studies demonstrated that both C5a and FMLP induce dose-related increases in CR1 (C3b receptor) and CR3 (iC3b receptor) expression in both monocytes and neutrophils.(ABSTRACT TRUNCATED AT 400 WORDS)     

Ticks (Acari: Ixodidae) parasitizing wild carnivores in Phu Khieo Wildlife Sanctuary, Thailand

Ixodid ticks were collected and identified from 8 wild carnivore species in Phu Khieo Wildlife Sanctuary, northeastern Thailand. Six tick species belonging to 4 genera were recovered and identified from 132 individuals. These included Amblyomma testudinarium (n = 36), Haemaphysalis asiatica (n = 58), H. hystricis (n = 31), H. semermis (n = 3), Rhipicephalus haemaphysaloides (n = 3), and Ixodes granulatus (n = 1). Leopard cats (Prionailurus bengalensis) (n = 19) were infested with 4 tick species, whereas yellow-throated marten (Martes flavigula) (n = 4), clouded leopard (Neofelis nebulosa) (n = 2), and dhole (Cuon alpinus) (n = 1) were infested with 3 tick species, Asiatic golden cat (Catopuma temmincki) (n = 2) with 2 species, and marbled cat (Pardofelis marmorata), binturong (Arctictis binturong), and large Indian civet (Viverra zibetha) each infested with 1 species. This information contributes to the knowledge available on the ectoparasites of wild carnivores in Southeast Asia.     

Relationships between complement activation, complement binding, and EBV absorption by human hematopoietic cell lines.

Complement consumption (C.C.) and C3 deposition on the cell membrane, visualized by membrane fluorescence (CMF), were compared in a collection of established human lymphoid lines. C.C. was independent of the presence of C3 receptors. A positive CMF reaction was seen only in lines that expressed C3 receptors, however. Trypsin treatment abolished CMF and EAC rosetting but had virtually no influence on C.C. EBV absorptive capacity correlated with both C3-receptor expression, as measured by EAC rosetting, and CMF but not with C.C. This is in line with our previous finding on the association of EBV and C3 receptors.