按不同直径分布的红细胞的变形性研究

目的：研究按不同直径大小分布的红细胞所对应的变形性及其膜的弹性模量。方法：将红细胞按直径大小分成六组，分别用底部附着法测出各组的应变量ε及膜的弹性模量E。结果：红细胞膜的弹性模量与直径大小呈正相关，大小红细胞之间弹性模量的变异量为86．76％。结论：小直径红细胞的变形性明显好于大直径的红细胞。     

Antibodies to Human Immunoglobulin Detected by Hemolysis of Human Immunoglobulin-Coated Red Blood Cells

A hemolysis assay was developed to detect alloantibodies to human immunoglobulin. A total of 1035 serum samples was tested. Anti-IgM antibodies were found in 8% of 59 normal persons and in 13% of 439 multiparous women, with the highest incidence of 67% in 341 dialysis patients. Although the anti-IgM antibodies were inhibited by both IgM and IgG, it appeared that they were also inhibited by F(ab')₂ but not by Fc. Anti-IgG antibodies were more strongly inhibited by Fc than F(ab')₂. These results suggest that anti-IgM antibodies might be analogous to antiidiotypic antibodies directed to F(ab')₂, whereas anti-IgG antibodies tend to have greater reactivity to Fc.     

Cytoadherence of Plasmodium berghei-Infected Red Blood Cells to Murine Brain and Lung Microvascular Endothelial Cells In Vitro

Sequestration of infected red blood cells (iRBC) within the cerebral and pulmonary microvasculature is a hallmark of human cerebral malaria (hCM). The interaction between iRBC and the endothelium in hCM has been studied extensively and is linked to the severity of malaria. Experimental CM (eCM) caused by Plasmodium berghei ANKA reproduces most features of hCM, although the sequestration of RBC infected by P. berghei ANKA (PbA-iRBC) has not been completely delineated. The role of PbA-iRBC sequestration in the severity of eCM is not well characterized. Using static and flow cytoadherence assays, we provide the first direct in vitro evidence for the binding of PbA-iRBC to murine brain and lung microvascular endothelial cells (MVEC). We found that basal PbA-iRBC cytoadherence to MVECs was significantly higher than that of normal red blood cells (NRBC) and of RBC infected with P. berghei K173 (PbK173-iRBC), a strain that causes noncerebral malaria (NCM). MVEC prestimulation with tumor necrosis factor (TNF) failed to promote any further significant increase in mixed-stage iRBC adherence. Interestingly, enrichment of the blood for mature parasites significantly increased PbA-iRBC binding to the MVECs prestimulated with TNF, while blockade of VCAM-1 reduced this adhesion. Our study provides evidence for the firm, flow-resistant binding to endothelial cells of iRBC from strain ANKA-infected mice, which develop CM, and for less binding of iRBC from strain K173-infected mice, which develop NCM. An understanding of P. berghei cytoadherence may help elucidate the importance of sequestration in the development of CM and aid the development of antibinding therapies to help reduce the burden of this syndrome.     

Autologous Red Blood Cells Potentiate Antibody Synthesis by Unfractionated Human Mononuclear Cell Cultures

We have tried to determine the most favourable conditions for the in vitro induction of specific antibody (Ab) responses to tetanus toxoid (TT) and keyhole limpet haemocyanin (KLH). Human peripheral blood mononuclear eells (PBMNC) were obtained from normal volunteers and stimulated with PWM, TT, KLH. and mixtures of PWM and antigens in the presence or absence of autologous red blood cells (RBC) (I:50 ratio of PBMNC/RBC). The cultures were harvested on day 11: immunogiobulins were determined immunonephelometrically and Ab levels by ELISA with human antibodies used for calibration. While anti-TT responses were easy to induce with PBMNC from recently boosted individuals, the production of anti-IT from PBMNC obtained from non-recently boosted individuals was only possible when PBMNC were stimulated with TT and PWM in ihe presence of autologous RBC Similarly, anti-KLH responses were easier lo induce with PBMNC frt)m an immune donor; maximal response was observed after slimuiation wiih PWM + KLH in the presence of autologous RBC. Stimulation of primary anti-KLH responses with PBMNC from non-immune donors was only successful when the cells were stimulated with KLH + PWM in the presence of autologous RBC. The potentiation of human B-cell responses with autologous RBC can be abrogated by pretreatment of PBMNC with anti-CD2 antibodies and is associated with increased expression of IL-2 receptors and increased production of gamma interferon (IFN-γ). However, addition of IFN-γ in different doses and at different times to PWM-slimulaled PBMNC cultures was not as effective as addition of RBC in enhancing the production of immunoglobulin and antibody.     

单克隆抗人B型红细胞抗体NAN123

NAN 123单克隆抗体能凝集人B型红细胞,凝集反应能被2 ~(-6)M的蜜二糖或 2 ~(-2)M的半和糖完全阻断。类似的结果在以CNE_2细胞为靶抗原的ELISA反应中也可观察到。实验提示,NAN123抗体对B血型抗原末端的α-D-半乳糖有选择性识别能力。     

Generation of Dendritic Cells from Peripheral Blood Adherent Cells in Medium with Human Serum

Dendritic cells (DC) provide an effective pathway for presenting antigens to T cells, both self-antigens during T-cell development and foreign antigens during immunity. As such, these cells may be promising adjuvants for immunotherapy. Thus, it is important to establish simple and fast method(s) to generate sufficient numbers of human DC in medium free of calf serum so that the cells can be used for both experimental and clinical purposes. In this report, we used peripheral blood adherent cells, without laborious cell purification or depletion, as the starting population and cultured them in medium supplemented with granulocyte/macrophage colony-stimulating factor and interleukin-4. Substantial numbers of cells with the phenotypical and functional characteristics of immature DC were obtained in a 7-day culture. We then compared DC cultured in medium supplemented with either fetal calf serum or pooled human ABRh⁺ serum and found no difference in cell yields and in their ability to stimulate alloreactive T cells or to present soluble antigens to T cells. Irradiated cells were less efficient than non-irradiated cells in antigen presentation and stimulation of T cells. Finally, we have examined DC with or without additional tumour necrosis factor-α treatment and found that antigen-pulsed mature cells could as efficiently present antigen to T cells as did immature cells. This method is suitable for the generation of DC in studies of large clinical materials.     

Potential of Laser Immunoassay for Detection of HIV in Human Blood Serum and Urine

Abstract

The potential of Light Scattering Immunoassay (LIA) for detection of HIV in human blood serum has been explored by monitoring the agglutination of antigen coated polystyrene particles by dynamic light scattering. Elisa tested human sera having HIV, TB, Filaria along with normal sera have been analyzed using two specific synthetic peptide antigen (SP1, SP2) and one nonspecific peptide antigen (NSP). Few paired human sera and urine samples and nonspecific (of nonHIV diseases) urine samples have also been tested using the same antigens to check the possibility of replacement of sera by urine.     

Human Lymphocyte Subpopulations: Rosette Formation with Sheep, Human and Horse Red Blood Cells

Rosette formation between human lymphocytes and horse red blood cells could be promoted by a low pH medium, overnight incubation and a temperature of 4°C. The percent of sheep, horse and human rosette-forming cells in the peripheral blood were 71.7 ± 1.8, 30.5 ± 2.8 and 28.3 ± 3.4 respectively. However, their percentages in thymuses were 97.1 ± 1.1, 91.4 ± 2.4 and 89.0 ± 3.4. Using preparations of isolated subpopulations, it was observed that the horse and human red cell rosette-forming cells were probably also “early” sheep red cell rosette-forming cells. Rosette formation with all three types of red blood cells were inhibited by a preparation of Fetuin-glycopeptide.     

老化红细胞膜表面抗原的研究

红细胞的老化是一个复杂的问题,对其老化机制争论较多,最近有人提出老化红细胞表面可结合自身IgG,然后被巨噬细胞及网织内皮细胞识别而清除,但缺乏直接的证据。本文用过氧化物酶标记Protein A与年轻及老化红细胞保温,老化红细胞结合的IgG是年轻者的两倍多。用FITC标记IgG抗体在流式细胞荧光检测仪测定,结果与酶标者一样。如用神经氨酸酶处理年轻红细胞,可提高酶标记及荧光标记的量,再用β-半乳糖苷酶水解,其标记量下降。实验结果说明老化红细胞表面确有IgG结合,其结合与半乳糖基有关。     

甲氧基聚乙二醇修饰人红细胞血型抗原的研究

红细胞输注是输血医学的主体，化学材料甲氧基聚乙二醇（mPEG）与红细胞膜共价结合后，可遮蔽红细胞表面血型抗原，使红细胞血型抗原呈阴性，即形成通用型红细胞，对临床输血具有重要意义。研究了mPEG修饰对红细胞血型抗原、膜流动性的影响；红细胞被修饰程度；mPEG与红细胞结合的稳定性；mPEG在体内的代谢情况。结果表明，mPEG能遮蔽红细胞上的血型抗原；mPEG与红细胞结合稳定；mPEG修饰红细胞的比率随浓度增高而增大；mPEG在小鼠体内24h后基本被代谢，且在体内无蓄积。mPEG修饰的红细胞为解决临床急用稀有血型和配型困难（如自身溶血性贫血，AIHA）等问题提供了有意义的参考。     

The Quality of Red Blood Cells

The evolving practice of medicine has required a number of changes in red cell product manufacture to ensure that the final product is more specifically tailored to the needs of the individual patient. As a result of the increasing concern over the risks of transfusion pharmaceutical standards of manufacture are now applied to blood component preparation. Studies have been undertaken to define the optimum method of blood processing, and newer technologies are emerging to allow acquisition of a more consistent dose of red cells in a fashion which may minimize the lesion of collection. Use of high efficiency 3+ generation filter technologies reduces leukokine build up during storage and improves the quality and purity of the stored blood product. The combination of new plasticizers for packaging and improved red cell additive solutions should allow the blood center to supply a more functional red cell with longer storage shelf life. Overall these developments should result in the provision of a more consistent dose of fully functional red cells to the recipient who will be less exposed to the undesirable sequelae of transfusion than previously.     

红细胞深低温保存方法的比较研究

探寻可替代甘油并简化复温后洗涤过程的较理想的红细胞低温保护剂,为低温保存红细胞临床应用提供便利。依据血红蛋白溶液浓度与其在波长为412nm处吸光度的关系曲线,建立红细胞回收率评估方法,研究比较了使用5种低温保护剂实现人类红细胞液氮深低温保存效果的优劣。结果表明:35.5%(w/v)甘油的保存效果最佳,其回收率为97.84%;其次为25%(w/v)右旋糖酐40,其回收率为85.08%;再次之为1M海藻糖(加载并孵育17.5h),其回收率为55.35%;回收率最低的两组为1M海藻糖(未经孵育)和10%(w/v)的二甲亚砜,对应的回收率均不足35%。初步推断:右旋糖酐40是一种可望替代甘油的红细胞低温保护剂。     

干预离子通道联合密度梯度离心富集胎儿有核红细胞的研究

 摘要:目的 探讨干预膜表面K-Cl 同向转运体（KCC）离子通道联合密度梯度离心对富集脐血中胎儿有核红细胞（NRBC）的效果。方法 应用KCC特异激活剂尿素（Urea）干预细胞膜,建立最大限度缩小细胞体积的干预条件;应用最佳干预条件干预脐血后进行不同密度介质下离心富集NRBC,用流式细胞仪计数观察胎儿NRBC的富集效果。结果 比较传统1.077淋巴细胞分离液离心后的单个核层流式细胞仪计数NRBC8.6%,干预脐血后用1.065密度介质离心后下层白膜层达69.9%,未干预组中用1.099密度介质离心的单个核层达55.9%,分别提高富集量8倍和6.5倍。结论 尿素干预联合密度梯度离心能有效提高NRBC的富集率。 关键词:胎儿有核红细胞;离子通道;尿素;干预;流式细胞仪     

Glycerol Transport in Human Red Cells

The kinetics of ¹⁴C-glycerol exchange was studied in human red cells. Glycerol appeared to be transported by two mechanisms: (i) by facilitated diffusion with permeability depending on glycerol concentration, and (ii) by an unspecific pathway, presumably representing the diffusion of individual glycerol molecules through the membrane with permeability independent of glycerol concentration. The latter permeability was 8 times 10^-8 cm/s at 20°C, it was independent of pH, and had an activation energy of 25 kcal/mol. The facilitated transport of glycerol was completely inhibited by Cu⁺⁺, and the activation energy was low, about 10 kcal/mol. The transport system was competitively inhibited by H⁺, reacting with at least three hydrogen ions. The chemical specificity of the transport system was remarkably low: 1,3 propandiol, a structural analogue, as well as dimethylsulfoxide (a hydrogen bonding molecule with no structural resemblance to glycerol), inhibited glycerol transport competitively. Steins “dimerizer hypothesis” was revised according to our findings. A kinetic scheme describing the reactions of a transport controlling site with glycerol is presented in the Appendix. It is demonstrated in the article that the scheme accounts for our experimental results.     

Modulation of the Interleukin 2 Receptor by Maternal and Cord Blood Serum

In this report, the potential inhibitive action, of maternal serum (retroplacental and peripheral), and cord blood serum on the expression of the I12r has been studied. Calf and male serum were used as a control. In order to have an optimal I12r expression, a previous PHA cellular stimulation was performed. the maternal serum's I12r inhibitive property was measured during and after the pregnant period. the presence of I12r on maternal lymphocytes, cord blood cells, and unrelated donor mononuclear cells has been investigated (after a PtfA stimulation assay). the down regulated Il2r expression of neoplastic cell line (Hut7g cell line) under the influence of maternal serum has been observed. the examination of the inhibitive action due to maternal serum has suggested that a factor included in the IgG fraction is mainly responsible for the down regulating property concerning the 112r expression. A possible mechanism for the action of this factor has been studied. Further experiments suggest that the addition of recombinant I12 during the action of low doses of maternal IqG allows a partial reexpression of the I12 receptor. However, at. physiological concentrations of IyC, thc Interleukin 2 receptor down regulation becomes Irreversible.