An enzyme-linked immunosorbent assay (ELISA) for detection of Fc alpha receptors on isotype-specific T cells

A sensitive and reproducible enzyme-linked immunosorbent assay (ELISA) has been developed using T cell hybridomas as coating antigen, for detection of Fc receptors for IgA (Fc alpha R). T-T hybridomas were generated from fusions of Fc alpha R+ T cell clones from mouse Peyer's patches with the Fc alpha R- R1.1 T lymphoma cell line. The 2 T-T hybridomas (designated Th HA) used here express Fc alpha R as determined by a rosette method and by ELISA. Th HA cells were cultured under conditions for maximum Fc alpha R expression, were added to individual wells of 96-well EIA plates, and were fixed in situ with glutaraldehyde. Plates were incubated with purified mouse monoclonal IgA, IgM or IgG1 and were developed with beta-galactosidase-coupled goat IgG antibodies specific for mouse heavy chains. Using the ELISA, both Th HA cell lines were shown to express significant levels of Fc alpha R, lower but detectable Fc mu R, and no discernible Fc gamma 1R. Interestingly, the rosette assay only allowed detection of receptors for IgA. When splenic lymphocytes were used, good Fc mu R and less Fc alpha R expression occurred on these cells as determined by ELISA and rosetting; however, no Fc gamma 1R cells were detected by either method. Thus, the ELISA is sensitive and reproducible, and allows an objective measurement of FcR expressed on T cells.     

Intense training: mucosal immunity and incidence of respiratory infections

This investigation examined the impact of a multistressor situation on salivary immunoglobulin A (sIgA) levels, and incidence of upper respiratory tract infection (URTI) during the French commando training (3 weeks of training followed by a 5-day combat course). For the URTI, the types of symptoms were classified according to the anatomical location of the infection. Saliva samples were collected (8 a.m.) from 21 males [21 (2) years] before entry into the commando training, the morning following the 3 weeks of training, after the 5-day combat course, and after 1 week of recovery. sIgA, protein and cortisol concentrations were measured. Symptoms of URTI were recorded during the study from health logs and medical examinations. After the 3 weeks of training, the sIgA concentration was not changed, although it was reduced after the 5-day course [from 120 (14) mg l^–1 to 71 (9) mg l^–1, P<0.01]. It returned to pre-training levels within a week of recovery. The incidence of URTI increased during the trial (²=53.48; P<0.01), but was not related to sIgA. Among the 30 episodes of URTI reported, there were 12 rhino-pharyngitis, 6 bronchitis, 5 tonsillitis, 4 sinusitis and 3 otitis. Cortisol levels were raised after the 3-week training (P<0.01), dropping below baseline after the combat course (P<0.01). Stressful situations have an adverse effect on mucosal immunity and incidence of URTI. However, the relationship between sIgA and illness remained unclear. The large proportion of rhino-pharyngitis indicated that the nasopharyngeal cavity is at a higher risk of infection.     

Probiotics in infancy induce protective immune profiles that are characteristic for chronic low-grade inflammation

Background Probiotics are widely studied both in the treatment and prevention of allergic diseases, but their mode of action is poorly known. Objective Our aim was to examine the effect of probiotic bacteria on in vivo cytokine, antibody, and inflammatory responses in allergy‐prone infants. Methods In a randomized double‐blind study, probiotic bacteria or placebo were given for 1 month before delivery to mothers and for 6 months to infants with a family history of allergy. Plasma samples were analysed for C‐reactive protein (CRP), total IgA and IgE, food‐specific IgA, IgG, and IgE, IL‐2, IL‐4, IL‐6, IL‐10, TNF‐α, and IFN‐γ. We analysed the associations of immunological and inflammatory parameters at age 6 months with probiotic treatment and allergic phenotype at 2 years. Results Infants receiving probiotic bacteria had higher plasma levels of CRP (P=0.008), total IgA (P=0.016), total IgE (P=0.047), and IL‐10 (P=0.002) than infants in the placebo group. Increased plasma CRP level at age 6 months was associated with a decreased risk of eczema [odds ratio (OR) 0.41 [95% confidence interval (CI) 0.17–0.99], P=0.046], and with a decreased risk of allergic disease [OR 0.38 (95% CI 0.16–0.87), P=0.023] at age 2 years, when adjusted with probiotic use. Conclusion The association of CRP with a decreased risk of eczema at 2 years of age in allergy‐prone children supports the view that chronic, low‐grade inflammation protects from eczema. Probiotic‐induced low‐grade inflammation was characterized by elevation of IgE, IgA, and IL‐10, the changes typically observed in helminth infection‐associated induction of regulatory mechanisms. The findings emphasize the role of chronic microbial exposure as an immune modulator protecting from allergy.     

An enzyme-linked immunosorbent assay (ELISA) for IgG and IgA antibodies to respiratory syncytial virus in low dilutions of secretions of human serum and secretions

An enzyme-linked immunosorbent assay (ELISA) has been developed for titration of IgG and IgA antibodies to respiratory syncytial (RS) virus in low dilutions of human serum, colostrum, and nasopharyngeal secretions. Previously the sensitivity of RS virus ELISA on such specimens has been limited by nonspecific absorption of antibody, particularly IgA, to crude antigen preparations. For IgG antibody estimation in infant sera, this unwanted binding was reduced to workable levels by increasing the serum, salt, and detergent concentration of the diluent. Residual nonspecific binding of IgA in colostra appeared mainly due to antigen lipids or to lipoproteins. This was markedly reduced by partitioning Triton X-100-treated infected cell lysate antigens in Arklone. Using the modified ELISA technique for anti-RS virus IgA, good correlations were found with unfixed cell membrane immunofluorescence (MIF) for colostra (r = 0.81, P less than 0.001) and nasal secretions from adult volunteers. In several samples nonspecific absorption of antibody precluded MIF assay, but did not affect the ELISA. Although there was an overall correlation between ELISA for anti-RS IgG antibody in sera, the complement fixation test (r = 0.75, P less than 0.001), and MIF test (r = 0.82, P less than 0.001), the sensitivity of ELISA for antibody responses in convalescent sera of infants from 3 months to 2 years was poor. Conversely, the sensitivity of ELISA for antibody in the sera of older children and for transplacentally acquired antibody in very young infants was higher than that for the other two tests. ELISA was thus less reliable than either CF or MIF for detecting antibody rises in paired infant sera, particularly where maternally acquired antibody remained in the acute serum. The reasons for this apparent disparity are discussed.     

《伤寒论》六经辨证在IgA肾病诊治中的指导价值

《伤寒论》六经辨证是在《黄帝内经》六经理论及脏象、经络两学说的基础上发展、完善而得来的,是中医的鼻祖。本文的主要目的是探讨《伤寒论》六经辨证在IgA肾病诊治中应用价值。IgA肾病又称为原发性肾小球疾病,在我国常见,其整个发病过程可视为从三阳经(太阳、阳明、少阳)转变为三阴经(太阴、少阴及厥阴)。IgA肾病在各经为病,脉症特点并不相同,因此IgA肾病采用六经辨证为指导,判断病情,能够有效防治IgA肾病,对临床具有重要的意义及价值。     

RTS,S/AS01E malaria vaccine induces IgA responses against CSP and vaccine-unrelated antigens in African children in the phase 3 trial

BackgroundThe evaluation of immune responses to RTS,S/AS01 has traditionally focused on immunoglobulin (Ig) G antibodies that are only moderately associated with protection. The role of other antibody isotypes that could also contribute to vaccine efficacy remains unclear. Here we investigated whether RTS,S/AS01_E elicits antigen-specific serum IgA antibodies to the vaccine and other malaria antigens, and we explored their association with protection.MethodsNinety-five children (age 5–17 months old at first vaccination) from the RTS,S/AS01_E phase 3 clinical trial who received 3 doses of RTS,S/AS01_E or a comparator vaccine were selected for IgA quantification 1 month post primary immunization. Two sites with different malaria transmission intensities (MTI) and clinical malaria cases and controls, were included. Measurements of IgA against different constructs of the circumsporozoite protein (CSP) vaccine antigen and 16 vaccine-unrelated Plasmodium falciparum antigens were performed using a quantitative suspension array assay.ResultsRTS,S vaccination induced a 1.2 to 2-fold increase in levels of serum/plasma IgA antibodies to all CSP constructs, which was not observed upon immunization with a comparator vaccine. The IgA response against 13 out of 16 vaccine-unrelated P. falciparum antigens also increased after vaccination, and levels were higher in recipients of RTS,S than in comparators. IgA levels to malaria antigens before vaccination were more elevated in the high MTI than the low MTI site. No statistically significant association of IgA with protection was found in exploratory analyses.ConclusionsRTS,S/AS01_E induces IgA responses in peripheral blood against CSP vaccine antigens and other P. falciparum vaccine-unrelated antigens, similar to what we previously showed for IgG responses. Collectively, data warrant further investigation of the potential contribution of vaccine-induced IgA responses to efficacy and any possible interplay, either synergistic or antagonistic, with protective IgG, as identifying mediators of protection by RTS,S/AS01_E immunization is necessary for the design of improved second-generation vaccines.Clinical trial registration: ClinicalTrials.gov: NCT008666191.     

Pathogenesis of idiopathic IgA nephropathy

Despite a prodigious amount of work on the physiology of IgA production in man, and many studies on the immunopathology of IgA nephropathy, ranging from the immunogenetics to the immune response to chemical characteristics of the IgA, we are hardly any nearer to defining the pathogenesis of this disease. One of the main changes in our understanding has been to recognise that the bone marrow, now known to produce normally one-third of the body's IgA, overproduces this immunoglobulin in IgA nephropathy. This alters the previous notion that IgA nephropathy was due simply to IgA production in the mucosa, although a mucosal component is not excluded. Certain characteristics of the IgA in the diseased kidney and the circulation have been defined: it is of subclass IgA1 and has a higher proportion of light chains and negative charge than in normal subjects. The specificities of the IgA, either in the kidney or in complexes, have not helped to clarify the pathogenesis. They have been found for a wide range of endogenous and exogenous antigens, suggesting that the antibody activity represents polyclonal B cell activation. These findings have not helped to confirm the prevailing theory that IgA nephropathy is an immune complex disease. Other theories put forward are that IgA nephropathy is an autoimmune disease, glomerular components or IgA itself being among the candidate antigens, or that there is primary dysregulation of the IgA immune system. At this stage of development in our understanding of this common nephropathy, it is important to guard against the assumption that idiopathic IgA nephropathy is one disease and is the result of a single pathogenetic mechanism.     

IgA肾病患者的扁桃体免疫组织化学观察

目的：了解ＩｇＡ肾病（ＩｇＡ－Ｎ）与扁桃体免疫异常的关系。方法：经肾穿刺活检确诊为ＩｇＡ－Ｎ３１例,以慢性扁桃体炎１６例作对照,扁桃体切除术后运用ＡＢＣ方法对扁桃体组织进行免疫组化标记。结果：两组扁桃体在ＩｇＡ、ＩｇＧ的表达上存在明显的区别,ＩｇＡ－Ｎ患者的扁桃体组织内产生ＩｇＡ的淋巴细胞数显著增加,结论：ＩｇＡ－Ｎ患者存在着扁桃体免疫异常,切除扁桃可预防抗原入浸,消除扁桃体来源的ＩｇＡ,减少循环中的ＩｇＡ免疫复合物形成,进一步使肾小球基底膜区的ＩｇＡ免疫复合物沉积减少。     

Modified sternal elevation for children with pectus excavatum

Objective To investigate the relationship between intestinal mucus IgA content and mucus barrier function after surface burns. Methods Detection of IgA content in mucus was performed by enzyme linked immunosorbent assay (ELISA) at different time points after burns.Bacterial adherence to cultured epithelial cells (IEC-6) in vitro using E.coli was assessed for each group. Results The intestinal mucus barrier function declined, parallel to a decrease in IgA content after surface burn in mice.In the normal control group, mucus IgA content was 2.32 Dλ, and 2.51, 1.76, 1.49, 1.06 Dλ at 0.5 h, 1 h, 6 h and 24 h after burn, respectively.Bacterial adherence rate was 0.53 in control group, and 0.46, 0.69, 0.58, 0.81 at 0.5 h, 1 h, 6 h, 24 h after burn, respectively. Conclusion The decrease of intestinal mucus IgA contents is one of the reasons why intestinal mucus barrier function declines after burns.     

Polymorphisms of TAP, LMP and HLA-DM genes in the Chinese

Ｔｒａｎｓｐｏｒｔｅｒａｓｓｏｃｉａｔｅｄｗｉｔｈａｎｔｉｇｅｎｐｒｏｃｅｓｓｉｎｇ(ＴＡＰ),ｌｏｗｍｏｌｅｃｕｌａｒｗｅｉｇｈｔｐｒｏｔｅａｓｏｍｅ(ＬＭＰ)ａｎｄＨＬＡＤＭａｒｅＭＨＣｃｌａｓｓⅡｌｉｋｅｇｅｎｅｓｗｈｉｃｈｈａｖｅｂｅｅｎｍａｐｐｅｄｂｅｔｗｅｅｎＨＬＡＤＰａｎｄＨＬＡＤＯｌｏｃｉｉｎｔｈｅｃｌａｓｓⅡｒｅｇｉｏｎｏｆｔｈｅｈｕｍａｎｍａｊｏｒｈｉｓｔｏｃｏｍｐａｔｉｂｉｌｉｔｙｃｏｍｐｌｅｘ1　ＴｈｅｓｅｇｅｎｅｓｄｉｆｆｅｒｆｒｏｍｔｈｅｃｌａｓｓｉｃｃｌａｓｓⅡｇｅｎｅｓ…