低血铅水平与免疫球蛋白和淋巴细胞相关性

目的：探讨低铅水平对IgA、 IgG 、 IgM、 C3、 C4、 B淋巴细胞的影响。方法用石墨炉原子吸收光谱法对39名印刷印染工龄5年以上工人进行血铅测定,另选择距污染源较远且无水系联系的,不受工业污染的,从未接触过重金属的为对照组,分别对两组用免疫比浊法进行（ IgA、IgG、 IgM、 C3和C4）5项测定,流式细胞仪检测血B淋巴细胞水平,全自动生化仪检测白细胞数、总淋巴细胞绝对数和百分比。结果高低铅两组IgA、 IgG 、 IgM、 C3水平比较差异有统计学意义（P＜0.05）,白细胞总数、淋巴细胞百分比均低于低铅组（P＜0.05）,总淋巴细胞百分比和NK淋巴细胞亚群的绝对数和百分比却低于低铅组（ P＜0.05）。结论高血铅对体液免疫功能有一定的影响。     

Physiological and Immunological Changes Associated with Oral Microbiota When Using a Thermoplastic Retainer

Background: The study examined the oral microbiota, physiological and immunological changes in patients using thermoplastic retainers during three months of use. Methods: The study included several steps. Firstly, 10 swabs were collected from the buccal and palatal surfaces of the teeth of the patients, approximately 2 mL of saliva was collected from the same patients and 2 mL of saliva was collected from 10 healthy people to measure the pH and secretory IgA level. This was followed by the isolation and identfication of the bacterial isolates in the patient samples. Then, isolate susceptibility toward chlorhexidine (CHX) and their adhesion ability to thermoplastic retainer surfaces was measured. In addition to that the study estimated the numbers of Lactobacillus and Streptooccus mutans colonies during three months and finally, a comparsion of pH acidity and IgA level between the patients and healthy people was performed. The results showed the predominant bacteria during the three months were Lactobacillus spp. and Streptococcus spp. followed by different rates of other bacteria. Raoultella ornithinolytica showed more resistance to CHX while Lactobacillus spp. showed more sensitivity. Streptococcus mutans colony levels were higher than Lactobacillus spp. colonies during the three months, also S. mutans had the highest value in adherence to retainer thermoplastic. Finally, pH acidity showed a highly significant difference (p ≤ 0.05) in the third month, like IgA levels (p ≤ 0.05). Conclusions: According to the results obtained from the current study, the researchers noted that the thermoplastic retainers helped change the oral cavity environment.     

Effects of peptide density and elution pH on affinity chromatographic purification of human immunoglobulins A and M

A family of linear hexamer peptide ligands HWRGWV, HYFKFD and HFRRHL, initially identified for their affinity to the Fc portion of human immunoglobulin G (hIgG), also have potential for use in the purification of human immunoglobulins A (hIgA) and M (hIgM). HWRGWV demonstrated the strongest binding affinity to hIgM, followed by hIgA and hIgG respectively. The effects of N-terminal acetylation of the peptide, as well as elution buffer pH, on the chromatographic elution of human IgG, IgA and IgM from HWRGWV resins at various peptide densities (0.04-0.55 meq/g) were investigated. Over 80% recovery and 90% purity were achieved for human IgG and IgA isolation from complete minimum essential medium (cMEM) using HWRGWV resin at optimum peptide densities. For human IgM, 75.7% recovery and 86.0% purity were achieved by using HWRGWV at a low peptide density of 0.04 meq/g. Although HYFKFD and HFRRHL exhibited their ability for isolation of human IgG, IgA and IgM from cMEM as well, HWRGWV is the best option among them for large-scale purification of human IgG, IgA and IgM based on conditions tested.     

IgA肾病免疫吸附剂的研究(Ⅱ)

通过1,4-丁二醇-二缩水甘油醚法活化琼脂糖载体,既起到了活化载体的作用,同时又由于该活化剂分子是长链结构而给载体提供了一定长度的手臂,有利于人免疫球蛋白G(IgG)大分子配基与载体的连接,用该方法制成的免疫吸附剂对高IgA肾病病人血清中IgA进行了吸附研究.通过体外实验条件的优化发现,最高可吸附60%的IgA,同CDI法比较(吸附率可达35%左右),该吸附剂具有较高的吸附效率.     

A simple and rapid competitive enzyme-linked immunosorbent assay (cELISA) for high-throughput measurement of secretory immunoglobulin A (sIgA) in saliva

A simple competitive enzyme-linked immunosorbent assay (cELISA) was established for rapid measurement of secretory immunoglobulin A (sIgA) in saliva. The method was based on competitive reaction between the immobilized IgA and free IgA in the solution for the limited amount of horseradish peroxidase-conjugated rabbit anti-human IgA. In comparison with the conventionally used Sandwich ELISA, the cELISA is simpler, low-cost, and shows better reproducibility since it is not affected by the variation of capture antibodies from different batches. The assay time was also significantly reduced from more than 5 h to less than 3 h. Different curve-fitting models were compared, among which the fully specified logit-log model gave the best results. The linear working range and limit of detection were found to be 0.1-100 μg mL⁻¹ and 0.05 μg mL⁻¹, respectively. Matrix effects of saliva samples were investigated and a reasonable range of dilution factors were proposed. The developed method offers a very practical approach for high-throughput measurement of sIgA in saliva samples.     

Mucosal dendritic cells shape mucosal immunity

Dendritic cells (DCs) are key modulators that shape the immune system. In mucosal tissues, DCs act as surveillance systems to sense infection and also function as professional antigen-presenting cells that stimulate the differentiation of naive T and B cells. On the basis of their molecular expression, DCs can be divided into several subsets with unique functions. In this review, we focus on intestinal DC subsets and their function in bridging the innate signaling and adaptive immune systems to maintain the homeostasis of the intestinal immune environment. We also review the current strategies for manipulating mucosal DCs for the development of efficient mucosal vaccines to protect against infectious diseases.     

Laboratorial diagnosis of paracoccidioidomycosis and new insights for the future of fungal diagnosis

Paracoccidioidomycosis (PCM) is the most prevalent mycosis in Latin-America. As for other mycosis, its importance of has been largely underestimated, partially due to the limited geographical distribution of the etiologic fungal agent (Paracoccidioides brasiliensis). However, the advent of AIDS and other immune suppressing conditions is creating an emergent need for improved diagnostic tests envisaging simpler, cheaper, faster and more sensitive and accurate detection of pathogenic fungi, especially those causing systemic and opportunistic diseases. Routine laboratorial diagnosis of PCM disease relies mainly on direct observation of the fungus. However, culture growing is slow and, too often, definite diagnosis can only be obtained at later growing stages. Immunodiagnosis is also widely employed, although usually cumbersome and complex. Enzyme-based immunoassays are more amenable to automation for high-throughput testing, but may lead to cross-reactivity with other fungi. Plus, molecular diagnosis relying on polymerase-chain reaction (PCR) and nucleic-acid hybridization, although still at early stages of application to routine diagnosis of P. brasiliensis, has triggered the development of techniques for its improved specific detection, thus contributing for epidemiological studies as well. In the future, microarrays and newer biosensing technologies, coupled to new bionanotechnological tools, will certainly improve diagnosis of PCM and other mycosis through very specific and sensitive pathogen biomolecular detection.     

双嘧达莫联合厄贝沙坦对IgA肾病的疗效及其安全性分析

目的探究双嘧达莫联合厄贝沙坦对Ig A肾病的疗效及其安全性。方法采取回顾性分析方法对遵义医学院第五附属(珠海)医院2013年7月—2016年7月Ig A肾病患者88例资料进行收集分析,随机分为单药组和联合组。单药组采用单纯厄贝沙坦治疗;联合组采用双嘧达莫联合厄贝沙坦治疗。比较两组患者Ig A肾病治疗效果;治疗前后24 h尿蛋白、血肌酐;不良反应发生率。结果联合组患者Ig A肾病治疗效果比单药组高,P0.05;治疗前两组患者24 h尿蛋白、血肌酐差异不显著,P0.05;联合组治疗后24 h尿蛋白、血肌酐比单药组好,P0.05;联合组不良反应发生率和单药组差异不显著,P0.05。结论双嘧达莫联合厄贝沙坦对Ig A肾病的疗效及其安全性良好,可改善肾功能,无明显副作用,值得推广。     

Improved Sensitive Sandwich Enzyme Immunoassay for Secretory Immunoglobulin a in Human Serum

Abstract

To overcome the IgA interference in enzyme immunoassay for serum secretory IgA (SIgA), a new specific, simple and more sensitive sandwich enzyme immunoassay, fully free of the serum IgA interference, was developed. SIgA standards or samples were added into the wells of polystyrene plate coated with rabbit IgG antibody to human secretory component. After incubation, the wells were washed and then, horseradish peroxidase-labeled Fab′ fragment of goat IgG antibody to human α-chain was added and incubated. The wells were washed again to remove the unbound labeled antibody, and the enzyme activity specifically bound to the well was assayed using 3,3′, 5,5′ - tetramethylbenzidine and H₂O₂ as substrate. The enzyme reaction was stopped by addition of 2M H₂SO₄. The SIgA concentration was determined from the absorbance at 450 nm. The minimun detectable sensitivity was 6ng/ml. The assay system had very good selectivity overcoming the interference of IgA. As the result of high sensitivity, only small amount of sample (2 μ1 for serum) was needed for analysis. In this assay, no cross reactivity was found with purified human IgG, mIgA, IgM or free secretory component (FSC). The recovery of SIgA mixed with human sera or biles was 99.6–108.1%. The coefficients of within-assay and between-assay variation were 5.8–9.3% and 6.2–9.2% respectively. It correlated well with a liquid phase competitive radioimmunoassay for human serum SIgA (r=0.96, n=30, P<0.0l). The level of SIgA in normal human serum was 8.04±3.60 (SD) μg/ml (n=117) and increased significantly in patients with choledocho- lithiasis (57.35±49.70 μg/ml, n=15, P<0.0l). SIgA concentrations in bile samples were also determined by the 2 4′ assay under the condition that FSC did not, interfere with the assay.     

反复呼吸道感染儿童发锌和血清IgA的分析

用火焰原子吸收法检测了反复呼吸道感染儿童头发中锌的含量，用放免法检测血清IgA的含量。结果表明，患病组儿童的发锌值和血清IgA值均低于正常组儿童（P＜0．05）。患儿经补锌治疗1年后，总有效率达82．1％，发锌及血清IgA含量均增加，表明缺锌所导致的IgA含量下降，呼吸道粘膜分泌IgA缺乏是儿童反复呼吸道感染的重要原因之一，补充锌可有效提高患儿的免疫力，治疗儿童反复呼吸道感染。