Immunoglobulin binding properties of the Prosorba immunadsorption column in treatment of rheumatoid arthritis.

Studies of the humoral effects of the Prosorba column were conducted in conjunction with the Phase 3 trial of Prosorba versus sham therapy for rheumatoid arthritis (RA). When perfused with normal human plasma in vitro, Prosorba bound predominantly IgG with a maximal capacity of approximately 462 g of Ig per Prosorba column, equal to about 1.5% of circulating IgG. Prosorba treatment did not alter the concentrations of albumin, IgG, IgM, and IgA in 3 RA patients, except for a small dilutional effect. Kinetic studies demonstrated that Prosorba removed IgG > IgM, IgA, and IgM rheumatoid factor (RF) during the initial moments of apheresis and almost exclusively IgM RF after 15 min. No net protein removal occurred at > or = 60 min. Mean values of circulating immune complexes (CICs) were not significantly decreased by 12 weekly treatments. Complement was activated by the apheresis system upstream of the Prosorba column without changing C3 or C4 levels. We conclude that the Prosorba mechanism of action in RA is not bulk removal of Ig, but might involve modification of the CIC repertoire and could include, but not be limited to, effects related to complement activation.     

Soluble interleukin-2 receptor: elevated levels in serum and synovial fluid of patients with rheumatoid arthritis

Soluble interleukin-2 receptor (sIL-2R) levels were quantitated in the serum and synovial fluid (SF) of patients with rheumatoid arthritis (RA) and degenerative joint disease (DJD). A sandwich immunoassay, employing two monoclonal antibodies against distinct epitopes on the IL-2R, was utilized for measurement. We found a striking elevation of sIL-2R in RA SF as compared with DJD SF (RA, 1319 +/- 135; DJD, 416 +/- 59; p less than 0.001). RA serum sIL-2R levels were also significantly elevated over DJD levels. There was no interaction between rheumatoid factor (RF) and sIL-2R. RA patients with elevated sIL-2R levels had significantly longer disease duration, higher c-reactive protein (CRP) levels in serum and SF, and higher RF levels in serum and SF. The groups were similar in regard to other laboratory variables. The presence of elevated levels of sIL-2R in RA serum and SF confirms the presence of a heightened immune reactivity and in vivo activation of lymphocytes in RA.     

IgM, IgG, and IgA rheumatoid factors in pigeon hypersensitivity pneumonitis

The association of rheumatoid factor (RF) and lung disease in several immunologically mediated conditions has suggested that it may be physiopathologically relevant. Since previous reports in hypersensitivity pneumonitis (HP) have dealt mainly with the immunoglobulin M (IgM) RF measurement, we studied such antibody activity in other immunoglobulins, to determine the IgG and IgA RF levels in pigeon-HP, and in asymptomatic breeders (AB) and rheumatoid arthritis (RA) as controls. RFs were measured in 35 HP patients, 41 AB, 31 RA controls, and 55 healthy donors by enzyme-linked immunosorbent assay (ELISA) using human or rabbit immunoglobulin G (IgG), anti-IgM, F(ab')2 of IgG, and IgA F(ab')2 conjugates. An affinity chromatography, fragment crystallizable (Fc) preparations of IgG, pepsin digestion, and Western blots were used to confirm RF specificity. We also evaluated anti-avian antibodies (AA) and cross-reacting antibodies. The HP group revealed positive IgM (51.4%), IgG (31.4%), and immunoglobulin A (IgA) (34.2%) RF tests, and these antibody values exceeded the AB reference levels (P<0.02). HP and RA showed a similar frequency and distribution of RFs. Possible immunoassay interferences were excluded. As in other immunologically mediated diseases, IgG and IgA RFs may play a pathogenic role in HP, amplifying the inflammatory reaction, immune-complex formation, and complement activation. IgM-RF producing cells that have been implicated in the presentation of self and foreign antigens, and T-cell activation might induce the isotype switching of RFs.     

神经梅毒患者血清及脑脊液免疫学指标诊断特征分析

目的探讨神经梅毒患者血清及脑脊液免疫学诊断特点。 方法选取2013年6月至2016年7月首都医科大学附属宣武医院收治的35例神经梅毒患者,其中32例患者行血清与脑脊液免疫学指标检测,回顾性分析35例患者的检查结果,应用Fisher精确检验比较血清组与脑脊液组IgA、IgM的差异,应用卡方检验比较血清组与脑脊液组IgG的差异,并对脑脊液寡克隆条带阳性检出率和脑脊液24 h IgG合成率进行分析。 结果32例神经梅毒患者检测血清免疫球蛋白,有53.13%(17/32)的患者IgG升高,6.25%(2/32)的患者IgA升高,0%(0/32)的患者IgM升高;脑脊液中,有84.38%(27/32)的患者IgG升高,100.00%(32/32)的患者IgA升高,90.63%(29/32)的患者IgM升高;血清与脑脊液IgA、IgM升高率比较,差异有统计学意义(P<0.01),IgG升高率比较,差异无统计学意义(χ²＝7.27,P>0.05)。25例神经梅毒患者行CSF寡克隆电泳,IgG寡克隆条带阳性率为100.00%(25/25),96.00%(24/25)患者脑脊液24 h IgG合成率升高。 结论神经梅毒临床表现多样,是易误诊的可治性疾病。血清及脑脊液IgG多表现为升高,脑脊液IgA、IgM多表现为升高而血清IgA、IgM多表现为正常,脑脊液寡克隆条带阳性,脑脊液24 h IgG合成率升高等免疫学特点对神经梅毒诊断有意义。     

IgG heavy-chain subclass typing of myeloma paraproteins by isoelectric focusing immunoblot analysis

The objective of this study was to develop a clinical laboratory method for subclass typing of human immunoglobulin G (IgG) paraproteins. Serum proteins were isoelectrically focused (IEF) in a mini-gel and passively blotted by capillary diffusion onto untreated nitrocellulose. Unreacted sites on the nitrocellulose were blocked with bovine serum albumin and the bound IgG was detected with peroxidase-conjugated anti-human IgG1-4 monoclonal antibodies from WHO/IUIS clones. The IEF immunoblot specificity was demonstrated by analysis of documented IgG, IgA, and IgM myeloma proteins of known subclass and light-chain composition. IEF immunoblots of sera from 18 myeloma patients who had an above-normal total IgG concentration produced IEF immunoblot patterns composed of five to 10 discrete bands (pI range 6.0 to 8.4). In contrast, no detectable IgG bands were observed with sera containing IgA and IgM paraproteins. The observed subclass frequencies of IgG paraproteins were 56% IgG1 (10/18), 28% IgG2 (5/18), 11% IgG3 (2/18), and 5% IgG4 (1/18). IEF immunoblot analysis permits the monitoring of changes in the pI and subclass of an IgG paraprotein over the course of a myeloma patient's therapy program.     

Simultaneous analysis of serum immunoglobulins in patients with M protein using cellulose acetate membrane isoelectric focusing.

We developed a method for the simultaneous analysis of microheterogeneity of human serum IgG, IgA, IgM, IgD, and IgE, and serum protein pattern using cellulose acetate membrane isoelectric focusing, and analyzed in 11 healthy subjects and 67 patients with M protein (17 cases of multiple myeloma [MM] and 50 cases of monoclonal gammopathy of undetermined significance [MGUS]). Using this method, bands indicating the microheterogeneity of each immunoglobulin could clearly be detected.Among healthy subjects, the detected IgG, IgA, and IgM bands did not vary, but the detected IgE and IgD bands did vary. Therefore, IgA, IgM, and IgG were selected for comparison of serum immunoglobulins in MM and in MGUS. In the IgA-type M protein group, normal IgM and IgG bands were decreased in MM patients compared to MGUS patients, while the M band and other bands were increased in MM patients compared to MGUS patients, but the differences between the two groups were not significant. In the IgG-type M protein group, normal IgM, IgA, and IgG were significantly decreased in MM patients compared to MGUS patients. We examined the changes in electrophoretic pattern in six MM patients and eight MGUS patients with IgA-type M protein after neuraminidase treatment. The width of the M band in MM patients with IgA-type M protein decreased with neuraminidase treatment. On the other hand, the width of the M band in MGUS patients with IgA-type M protein increased with neuraminidase treatment. We concluded that the decrease of the normal immunoglobulins in MM patients with IgG type M protein could be detected by this method, and IgA type of M protein binding sugar chain were different between MM and MGUS patients.     

Cellular basis of hyper IgM immunodeficiency

Six patients with primary hypogammaglobulinaemia and hyper IgM were studied. All showed very low serum IgG and IgA concentrations. The in vitro pokeweed-mitogen (PWM)-induced immunoglobulin (Ig) production, including IgM, by their peripheral blood lymphocytes was low. Even when patients' B cells were cocultured with normal T cells, IgM production did not reach normal levels. These results and studies of Ig class on the surface of B lymphocytes point to a maturation arrest of these cells. T cells from all but one patient helped very little Ig production by patients' or normal B cells. Similar numbers of these T cells did not suppress Ig production by normal T plus B cells. Therefore a defect in T cell help for IgM, IgG and IgA was seen in most patients, in addition to the B cell abnormality.     

类风湿关节炎患者血清IgG及亚类的水平和临床意义

目的探讨类风湿关节炎(RA)患者血清IgG及亚类的水平和临床意义。方法选择2018年10月至2019年6月该院风湿免疫科收治的RA患者54例纳入RA组,选择同期36例健康体检者纳入对照组。采用双抗体夹心法ELISA检测血清IgG亚类水平;采用免疫散射比浊法检测免疫球蛋白(IgG、IgA、IgM)、补体(C3、C4)和类风湿因子(RF)水平;采用流式点阵免疫发光法检测抗环瓜氨酸肽(CCP)抗体水平。比较两组各检测指标水平,分析RA患者血清IgG亚类与IgG、IgA、IgM、C3、C4、RF及抗CCP抗体之间的相关性。结果RA组血清IgG、IgA、RF和抗CCP抗体水平显著高于对照组(P<0.05),但IgM、C3和C4水平差异无统计学意义(P>0.05)。RA组血清IgG1和IgG3水平显著高于对照组(P<0.05),两组血清IgG2和IgG4水平差异无统计学意义(P>0.05)。与对照组比较,RA组IgG1/IgG和IgG3/IgG显著升高(P<0.05),而IgG2/IgG显著下降(P<0.05)。Spearman相关分析显示,RA组患者血清IgG1水平与IgG呈高度正相关(r=0.865,P<0.05);IgG2、IgG3水平与IgG均呈中度正相关(r=0.613、0.644,P<0.05);IgG4水平与IgG呈低度正相关(r=0.271,P<0.05);IgG2水平与IgA呈低度正相关(r=0.399,P<0.05);IgG3水平与IgM呈低度正相关(r=0.343,P<0.05)。IgG各亚类与RF、抗CCP抗体之间均无相关性(P>0.05)。结论RA患者血清IgG水平显著升高,并且存在IgG亚类水平变化。IgG亚类检测对RA早期诊断价值有限。     

Immunoglobulin classes of DNA binding activity in serum and skin in systemic lupus erythematosus.

36 systemic lupus erythematosus patients with native DNA binding activity (nDNA-BA) in the serum and subepidermal immunoglobulin deposits were studied to determine the relationship of the immunoglobulin (Ig) class distribution of serum nDNA-BA to the clinical characteristics of their disease and to the Ig class present at the dermal-epidermal junction (DEJ). The patients with predominantly (86-98%) IgM nDNA-BA in the serum had less active disease, mild or no renal involvement, and longer survival than those with predominantly (51-95%) IgG nDNA-BA in the serum. Renal biopsies in eight patients with predominantly IgM nDNA-BA in the serum showed relatively benign histologic changes in the kidney. In contrast, renal tissue from 23 patients with predominantly IgG nDNA-BA showed more severe histologic changes. All patients had multiple skin biopsies. Patients with predominantly IgM nDNA-BA consistently had only IgM at the DEJ. Patients with predominantly IgG nDNA-BA had IgG, usually in association with IgM, at the DEJ. The findings demonstrate that a minority of systemic lupus erythematosus patients may exhibit a limited anti-nDNA response characterized by the presence of chiefly IgM nDNA-BA in the serum and that this is reflected by the presence of mild disease and IgM alone at the DEJ. The development of IgG nDNA-BA is associated with more severe and active disease.     

Decreased cytoplasmic immunoglobulin a production during epstein-barr virus immortalization on lymphocytes from patients with ataxia-telangiectasia

Previously, we have reported significantly lower immunoglobulin (Ig) A production in supernatants of cultured lymphoblastoid cells using enzyme-linked immunosorbent assay from patients with ataxia-telangiectasia (AT) when compared to that of age- and sex-matched healthy individuals. Here, we further assess the degree of cytoplasmic Ig production in these cells and also analyze it during the early phase of Epstein-Barr virus immortalization. All classes of cytoplasmic IgM, IgG, and IgA productions were demonstrated in cells from healthy controls. In contrast, cells from patients with AT showed only cytoplasmic IgM and IgG with low or nondetectable levels of IgA during and after the immortalizing process. These results suggest B lymphocytes bearing IgA are functionally immature and/or defective in patients with AT.©1995 wiley-Liss, inc.     

Monoclonal and oligoclonal gammopathies in heart-transplant recipients

Immunoglobulin abnormalities in serum from 76 heart-transplant recipients were examined by cellulose acetate and agarose gel electrophoresis. Monoclonal components were typed by immunofixation. IgG, IgA, and IgM and total kappa and lambda light chains were quantified by immunonephelometry. We confirm that both monoclonal and oligoclonal immunoglobulin banding are common in serum from these patients. Of the 149 serum samples examined, 21 (15%) had one monoclonal component and 53 (35%) had two or more. These monoclonal immunoglobulins were generally present at a low concentration and were transient. The class of immunoglobulins most commonly involved was IgG (about sevenfold more numerous than IgM); monoclonal IgA components and free light chains were not detected. The nephelometric kappa/lambda and heavy chain/light chain ratios were poor indicators of these abnormalities. Immunoglobulin abnormalities were not correlated with the sex and age of recipients, the pre-existing cardiopathy, the time since transplantation, or plasma concentrations of cyclosporine, but did correlate with plasma immunoglobulin concentration, biopsy findings, and viral infections, especially cytomegalovirus (CMV). A monoclonal IgG purified from a patient with a high titer of anti-CMV antibodies did not react with CMV antigens. The origin of these immunoglobulin abnormalities is unclear. Our data suggest that the presence of monoclonal or oligoclonal banding in heart-transplant recipients is of limited prognostic significance.     

自身免疫病患者碱性磷酸酶测定的意义

检测了50例自身免疫病患者的血清ALP、免疫球蛋白、补体及ENA多肽抗体。结果显示与对照组相比,自身免疫病患者的ALP活性升高(P＜0.05),IgG显著升高(P＜0.01),IgA、IgM升高(P＜0.05),C     

Ig同型转换及寡克隆蛋白条带在免疫固定电泳图谱中的形态特点及其对多发性骨髓瘤患者骨髓移植疗效评价的意义

目的探讨Ig同型转换及寡克隆蛋白条带在免疫固定电泳(IFE)图谱中的形态特点及其对多发性骨髓瘤(MM)患者骨髓移植疗效评价的意义。方法选取2004~2014年本院收治的MM患者64例,比较患者行骨髓移植前后血清蛋白电泳及IFE图谱资料,并进行分析与统计。结果 64例MM患者中,48.4%出现Ig同型转换和(或)寡克隆蛋白条带。异常蛋白条带(APB)首次出现中位数时间为移植后1.8(0~29)月。血清IFE结果提示71.0%的患者首先出现的是IgG kappa同型转换,IgG lambda占9.7%,IgA kappa占3.2%,IgA lambda占9.7%,IgM kappa占3.2%,LC占3.2%。移植后首先出现是以IgG kappa同型转换为多见。出现APB的患者与非APB患者无进展生存率分别为43%和0,差异有统计学意义(P=0.049);完全缓解率分别为86.7%和61.5%,差异有统计学意义(P=0.03)。至随访结束,31例APB患者与33例非APB患者比较5年生存期,分别为83%和57%,差异无统计学意义(P=0.12)。结论 MM治疗过程中出现APB往往提示疗效更高,预后更好。对MM治疗过程中新出现APB特点的解释及正确认识也将影响疗效评价的正确判断。     

Class- and subclass-specific antibody response to hemocyanin in nonmalignant paraproteinemia.

IgM, IgG, and IgA class-specific, as well as IgG subclass-specific antibody titers against the primary immunogen HPH were measured with ELISA in 19 patients with nonmalignant paraproteinemia (eight with IgG1, two with IgG2, two with IgG4, four with IgM, and three with IgA) and in a simultaneously studied age- and sex-matched control group. After primary immunization only IgM and IgA anti-HPH titers were significantly lower in the patient group. Four patients with relatively high IgG or IgA serum paraprotein levels did not produce antibodies in some Ig classes or IgG subclasses, whereas all other patients and all controls developed antibody titers in all classes and IgG subclasses. Low or absent antibody titers did not occur preferentially in the Ig (sub)classes to which the paraproteins belonged. After secondary immunization the patients could not increase or maintain their antibody titers as well as the controls, and this was most clear in the IgM and IgA antibody class. A direct correlation between polyclonal serum IgM levels and IgM anti-HPH titers was present in the patients. Such a correlation was absent for IgA in the patients and for all classes in the controls. It is concluded that humoral immunosuppression as measured with a newly encountered antigen in patients with nonmalignant paraproteinemia is most clearly expressed in the IgM and IgA antibody class and that the paraprotein (sub)class is not preferentially involved.     

免疫球蛋白及血细胞在肝肾不足、风湿阻络型尪痹中的表达

目的:探讨免疫球蛋白和血细胞与肝肾不足、风湿阻络型类风湿性关节炎(尪痹)的相关性?椒ǎ?随机选择符合要求的病例30例,测定免疫球蛋白和血细胞的值,并对所测结果进行统计学分析。结果:肝肾不足、风湿阻络型尪痹的免疫球蛋白中IgG、IgM、IgA显著升高(P<0.01),血细胞中HGB、PLT显著下降(P<0.01),WBC、C3无改变(P>0.05)。结论:IgG、IgM、IgA升高,HGB、PLT降低对肝肾不足、风湿阻络型尪痹有诊断意义,RBC对其诊断有参考意义,WBC、C3无诊断学意义。     

The class of surface immunoglobulin on cells carrying IgG memory in rat thoracic duct lymph: the size of the subpopulation mediating IgG memory.

The fluorescence activated cell sorter was used to determine the class of immunoglobulin on the thoracic duct lymphocytes (TDL) which carried IgG memory. Although only about 3% of all TDL carried membrane IgG these cells accounted for most, if not all, of the adoptive IgG anti-DNP response. It is concluded that both CR+ and CR- cells mediating IgG memory in rat TDL bear the same class of membrane immunoglobulin as that secreted by their differentiated progeny. The class of membrane immunoglobulin on CR+ and CR- rat TDL was also examined. It was found that IgM+ cells, which made up over 80% of all Ig+ cells, were virtually all CR+. In contrast, the few percent of IgG+ and IgA+ cells present were to be found in both subpopulations. There was no evidence of a large population of B cells bearing exclusively heavy chains other than IgA, IgG, of IgM. The observation that some IgG+ cells as well as IgM+ cells possess a receptor for C3 appears to rule out the hypothesis that this receptor is involved in blocking a switch from IgM to IgG synthesis.     

Oligoclonal immunoglobulin M in the cerebrospinal fluid of patients with Garin-Bujadoux-Bannwarth meningopolyneuritis

Paired cerebrospinal fluid (CSF) and serum samples from 15 patients with meningopolyneuritis Garin-Bujadoux-Bannwarth (MPN-GBB) were investigated by agarosegel electrophoresis (AE) and consecutive immunofixation (IF). Oligoclonal immunoglobulin (Ig) was detected in the CSF in 13 cases, 8 of which showed oligoclonal IgM; oligoclonal Ig was not found in the respective serum samples. Local CSF synthesis of IgM is a characteristic feature in patients with MPN-GBB. Further serological testing for borreliosis is mandatory in such cases.     

Antibodies to cyclic citrullinized peptide in rheumatoid arthritis

AIM: To estimate a diagnostic value of antibodies to cyclic citrullinized peptide (CCP) in rheumatoid arthritis (RA). MATERIAL AND METHODS: The study was made of 85 RA patients. Of them, 48 patients had early RA, i.e. of 8 month and less duration. The control group consisted of 35 patients with non-differentiated arthritis (NDA) and 8 healthy donors. Concentrations of CCP antibodies, rheumatoid factor (RF) IgM and RF IgA were measured with enzyme immunoassay (EIA). RESULTS: The level of CCP antibodies in RA patients (76.3 +/- 43.8; median 100.0 U/ml) was significantly higher than in NDA patients (25.1 +/- 43.9; median 0.8 U/ml; p < 0.05) or in donors (0.38 +/- 0.36; median 0.2 U/ml; p < 0.05). A correlation was found between the CCP antibodies level and that of RF IgM (chi2 = 15.4; p = 0.001) and RF IgA (chi2 = 10.3; p = 0.001). Sensitivity (82%) and specificity (90%) of CCP antibodies in RA diagnosis was higher than these parameters for RF IgM and IgA (78%, 86% and 72%, 83%, respectively). Simultaneous tests for CCP antibodies, RF IgM and RF IgA led to a 93% specificity. CCP antibodies were detected in 50% patients seronegative by RF IgM and in 62% patients seronegative by IgA. Detection of CCP antibodies was closely associated with early RA (chi2 = 30.8; p = 0.0001). CONCLUSION: The EIA for CCP antibodies is a sensitive and specific serological test for early RA diagnosis.     

The effect of anesthesia on the immunoglobulin level of the blood

The effect of halothane anesthesia, neuroleptanalgesia and ketamine anesthesia on immunoglobulin (Ig) blood level has been studied in 67 patients with gastric and duodenal ulcers subject to selective proximal vagotomy. The results obtained indicate possible reduction in humoral immunity under the influence of anesthesia and surgery. In patients operated on under halothane anesthesia there was a decrease in IgM, IgG and IgA levels. Neuroleptanalgesia produced a drop in IgA level, while ketamine anesthesia caused no significant changes in IgG blood level.     

Temporal invariance and clonal uniformity of brain and cerebrospinal IgG, IgA, and IgM in multiple sclerosis

Elevated cerebrospinal fluid (CSF) IgG and oligoclonal IgG bands on electrophoresis are valuable clinical markers for B cell proliferation in the brains of patients with multiple sclerosis (MS). Using two-dimensional electrophoresis, (2DE) we have established that the humoral immune response in MS brain is characterized by finite clonal complexity for the major Ig classes. An important question is whether this immune response is clonally stable or varies with time, related to the development of new lesions and random entry of B cells into the MS brain. To investigate this, we performed serial electrophoretic studies on CSF obtained from 19 patients with MS; the intervals ranged from 7 to 12 yr, with a mean of 8 yr. These analyses included studies of IgG, IgA, and IgM, and revealed that the humoral immune response in MS is clonally stable over long periods. Spontaneous fluctuations or reduction in CSF IgG levels by drugs did not qualitatively affect B cell clonal proliferation in MS brain, in that dominant bands and spots were not obliterated. It has been asserted that IgG synthesis in MS is nonsense antibody because the spectotypes of IgG isolated from different regions of MS brains differ. Factors other than clonal heterogeneity could account for differences found using one-dimensional analysis. B cell clonal products resolve into unique and well-resolved spots by 2DE; the method is uniquely suitable for analysis of restricted immune responses. Therefore, Ig were isolated from 11 regions of three MS brains and the 2DE patterns were compared. The similarity of the 2DE patterns indicate unequivocally that major clones are distributed uniformly although some clones are more prominent in some brain areas. IgA and IgM isolated from the same areas also showed similar patterns. Furthermore, the patterns of light and heavy chains in brain regions differed from serum but were similar to the autologous CSF, providing new evidence that CSF IgG in MS derives from synthesis in situ. Our results indicate that, once initiated, B cell clonal proliferation persists indefinitely and is little altered qualitatively at a clonal level over time, even when CSF IgG levels change or are altered by drugs. Our results are consistent with allotype and idiotype analysis of Ig production in MS and conflict with nonsense antibody proposals of the origin and nature of in situ synthesized Ig in MS.