抗CCP抗体、抗MCV抗体和类风湿因子在早期类风湿关节炎中的诊断价值

目的探讨抗CCP抗体、抗MCV抗体和类风湿因子在早期类风湿关节炎中的诊断价值。方法选择于2016年1月至2017年1月期间在我院就诊的早期类风湿关节炎(RA)患者及其他骨关节病患者各40例,并选择40名同期在我院体检的健康人作为研究对象。其中风湿关节炎患者设为RA组,其他骨关节病患者为疾病对照组,健康人为正常对照组。采用免疫透射比浊法、酶联免疫吸附法(ELISA)、免疫散射比浊法分别测定三组的抗环瓜氨酸肽(CCP)抗体、抗突变型瓜氨酸波形蛋白(MCV)抗体及IgM型类风湿因子(IgM-RF)水平,比较抗CCP抗体、抗MCV抗体、IgM-RF单独检测及三个指标联合检测对早期类风湿性关节炎的诊断价值。结果①RA组的抗CCP抗体、抗MCV抗体、IgM-RF水平均明显高于疾病对照组与正常对照组,差异具有统计学意义(P<0.05);疾病对照组与正常对照组间差异无统计学意义(P>0.05)。②RA组的抗CCP抗体、抗MCV抗体、IgM-RF及联合检测的阳性率均明显高于疾病对照组与正常对照组,差异具有统计学意义(P<0.05);疾病对照组与正常对照组间差异无统计学意义(P>0.05)。③联合检测的灵敏度及阴性预测值均明显高于抗CCP抗体、抗MCV抗体、IgM-RF单独检测,差异具有统计学意义;抗CCP抗体与抗MCV抗体的特异性及阳性预测值均明显高于IgM-RF及联合检测,差异具有统计学意义(P<0.05)。结论早期RA患者的血清中存在抗CCP抗体、抗MCV抗体、IgM-RF的高表达,且其灵敏度及特异性均较高,临床上可联合检测抗CCP抗体、抗MCV抗体及IgM-RF,以提高RA的早期诊断率。     

Kinetics, Localization and Isotype Profile of Antibody Responses to Immune Stimulating Complexes (Iscoms) Containing Human Influenza Virus Envelope Glycoproteins

The immune stimulating complex (iscom) is a particulate adjuvant formulation combining multimeric presentation of antigen with a built-in adjuvant, Quillaja saponin. Iscoms induce strong serum antibody responses that are readily boosted. To further characterize this property of iscoms, the development and maturation of primary and secondary antibody responses to iscoms containing influenza virus antigen were investigated, in serum by ELISA and on single B-cell level by ELISPOT. After a single subcutaneous injection, B cells secreting antigen-specific IgG (IgG-SC) were primarily observed in the draining lymph nodes (LN), showing peak numbers at day 7 which then declined rapidly. Serum IgG levels, as well as IgG-SC in the spleen, persisted for several weeks and, with time, IgG-SC cells also appeared in the bone marrow (BM). These results suggest that the IgG response to iscoms initially is located to the LN but that IgG-SC are redistributed with time and may persist for a long time in other organs, including the spleen and BM. Moreover, a booster dramatically enhanced the frequency of IgG-SC in LN, spleen and BM suggesting that iscoms induce a potent B-cell memory. Comparisons of antibody responses to iscoms with those to influenza virus antigen in Freund's complete adjuvant, TiterMax^TM or aluminium hydroxide suggest that the choice of adjuvant influences both the magnitude, kinetics, localization and isotype profile of antibody responses.     

Cell-Mediated Immunity in Humans During Viral Infection: Dermal Hypersensitivity and In Vitro Lymphocyte Proliferation During Mild Viral Respiratory Infections

In vitro lymphocyte proliferation in response to phytohemagglutinin and streptokinase-streptodornase and delayed dermal hypersensitivity to several antigens were assessed in patients with mild viral upper respiratory infections. The response to phytohemagglutinin in 18 patients was not diminished during the viral infection. Deoxyribonucleic acid synthesis induced by streptokinase-streptodornase remained unaffected by the viral illness in five patients, and skin test reactivity was not depressed during the viral infection. Thus, it appeared that localized viral upper respiratory infections were not associated with suppression of systemic cell-mediated immunity.     

Increased Production of a Th2 Cytokine Profile by Activated Whole Blood Cells from Rheumatoid Arthritis Patients

T cells produce regulatory cytokines which control inflammation. In rheumatoid arthritis (RA), a Th1 cytokine profile has been described in the synovium. In order to assess the Th1/Th2 cytokine balance in blood, a one step culture-immunoassay procedure was used to measure the ex vivo production of IFN and IL-4 by whole blood cells from 26 RA patients and 25 controls. For comparison, the same cytokines were measured by ELISA in supernatants of activated whole blood cells. The direct whole blood assay was 10-fold more sensitive than standard ELISA to measure IL-4 levels. IL-4 production was higher in RA patients than in controls, whereas that of IFN was lower. Accordingly, the IL-4/IFN ratio, which reflects the Th2/Th1 cytokine balance in blood, was higher in RA patients (P < 0.0001). The present findings indicate a Th2-over-Th1 cytokine balance profile in RA blood. These results are in contrast with the Th1-over-Th2 pattern previously found in the joint, indicating that the two compartments appear to be differently regulated.     

Development of Antibody Isotype Responses to Schistosoma mansoni in an Immunologically Naive Immigrant Population: Influence of Infection Duration, Infection Intensity, and Host Age

We have identified the influence of host and parasite factors that give rise to characteristic antibody isotype profiles with age seen in human populations living in different areas of schistosomiasis endemicity. This is important in the immunobiology of this disease. It is also of interest in the context of human responses to chronic antigen stimulation, vaccines, allergens, and other pathogens. In populations exposed to endemic schistosomiasis, factors such as intensity and duration of infection are age dependent. They therefore confound the influence of host age on antiparasite responses. Here, we resolved these confounding factors by comparing the developing antibody responses of an immunologically naive immigrant population as they acquired the infection for the first time with those of chronically infected resident inhabitants of the same region of Schistosoma mansoni endemicity in Kenya. Recent arrival in the area strongly favored immunoglobulin G3 (IgG3) responses against the parasite. The antibody isotype responses associated with human susceptibility to reinfection after chemotherapy were elevated in those suffering high intensities of infection (IgG4 responses against worm and egg antigens) or were characteristic responses of young children irrespective of the intensity or duration of infection (IgG2 responses against egg antigen). IgE responses against the adult worm, a response associated with resistance to reinfection after chemotherapy, increased with the ages of infected individuals and were also favored in those currently suffering higher intensities of infection.     

Comparison of Three Anti-CCP Antibody Tests and Rheumatoid Factor in RA and Control Patients

It was the aim of this study to compare the diagnostic properties of two second generation anti-CCP antibodies assays (Quanta Lite CCP, Inova and EliA CCP, Phadia) with an assay containing a new antigen mixture (Quanta Lite CCP 3, Inova) and RF IgM. METHODS: Sera from 86 RA patients and 90 control patients with other diseases, such as infections, connective tissue diseases, or other rheumatic diseases, were analysed using the four different methods. Specificity (84.4%) and PPV (80.3%) were lowest for RF IgM and highest for EliA CCP (specificity 97.8%, PPV 96.7%), the other values being close to those of EliA CCP. Sensitivity was highest for Quanta Lite CCP (73.3%). Efficiency was highest for Quanta Lite CCP (84.1%) at the lower cut off followed by EliA CCP at both cut offs (83.0%). The discrimination between RA sera (mean value 407.67 U/ml) and control sera (4.54 U/ml) and the relative risk (23.34) were best for EliA CCP. The results of this study reveal that there was no advantage of the anti-CCP antibodies assay applying a new antigen mixture (Quanta Lite CCP 3 ELISA) compared to two second generation anti-CCP antibodies assays (Quanta Lite CCP ELISA and EliA CCP).     

地高辛单克隆抗体的研究 Ⅰ、分泌地高辛特异性单克隆抗体杂交瘤细胞系的建立及抗体特性分析

地高辛(Digoxin)是目前应用最广泛韵强心甙类药物,在心血管疾病临床治疗中占有十分重要的地位。强心甙治疗的安全范围较小,一般治疗量已接近中毒量的60%,并且有较大的个体差异,使得临床应用时中     

The Distribution and Abnormal Morphology of Plasma Cells in Rheumatoid Synovium

This study assessed the distribution and structural features of plasma cells in rheumatoid synovial tissue. Plasma cells were found to be the predominant infiltrating mononuclear cells (mean 40%) in relation to lymphocytes and monocytes, and there was a direct relationship between their number in the infiltrates and the total number of mononuclear leucocytes (P = 0.007). Plasma cells were also seen in intimate contact with macrophages intermixed with synovial lining cells, and closely associated with small blood vessels. They often surrounded these blood vessels and sometimes were seen lying within the vessel walls themselves. Ultrastructural analysis revealed that many synovial plasma cells were considerably larger than plasma cells of a normal size and possessed a marked distension of the cisternae of rough endoplasmic reticulum. Furthermore, plasma cells in close proximity to blood vessels often appeared to be undergoing migration. These observations imply that in rheumatoid synovium, plasma cells are metabolically very active and occupy a pivotal position for the secretion of antibodies into both the vascular and the extravascular compartments.     

抗CCP抗体、RF与CRP联合检测在类风湿关节炎诊断中的意义

目的探讨血清中抗环胍氨酸肽抗体(A-CCP)、类风湿因子(RF)与C反应蛋白(CRP)联合检测在类风湿关节炎(RA)早期诊断中的意义。方法选取2016年10月至2018年5月在我院就诊的RA患者110例(RA组),同时选取其他风湿病患者97例(非RA组),健康体检者95例(健康组)的临床样本为试验材料。采用北京九强试剂定量检测A-CCP、RF及CRP水平,比较3种指标在类风湿关节炎(RA)早期诊断中的价值。结果RA组A-CCP、RF及CRP的阳性率显著高于非RA组及健康组,三者联合检测的灵敏性和特异性与单独检测时比较,差异有统计学意义(P<0.05)。结论抗CCP抗体、RF和CRP三者联合检测可大大提高早期诊断RA的灵敏度和特异性,对早期诊断RA具有重要意义。     

Antibody Response and Acquired Tolerance of A/J Mice: Age- and Immunogen-Related Isotype Differences

In earlier work we have observed strain- and age-related diversity of immunological ageing in inbred mice pretreated with tolerogen or left without pretreatment and then immunized. Different isotypes show different age- and strain-related changes. Here, we extend this isotype analysis from C57BL/6 and SJL to A/J. By comparison with the first two of these, animals of strain A/J show very little age-related change, as judged by indirect plaque-forming response. We have found that in A/J, as in SJL and C57BL/6 mice, age-related changes are isotype-dependent. The age-related changes in isotype predominance and magnitude differ for different determinants. They depend on the structural relation between the tolerance-inducing or -sensitizing macromolecule and the immunogen.     

During Trypanosoma cruzi Infection CD1d-Restricted NK T Cells Limit Parasitemia and Augment the Antibody Response to a Glycophosphoinositol-Modified Surface Protein

Trypanosoma cruzi is a protozoan parasite that chronically infects many mammalian species and in humans causes Chagas' disease, a chronic inflammatory disease. The parasite expresses glycophosphoinositol (GPI), which potently stimulates interleukin 12 (IL-12) production. During T. cruzi infection IL-12, and possibly GPI, might stimulate NK T cells to affect the protective and chronic inflammatory responses. Here we report that during T. cruzi infection CD1d-restricted NK T cells are stimulated as NK T-cell-deficient mice have greater parasitemia. Furthermore, during T. cruzi infection the percentages of NK T cells in the liver and spleen become decreased for prolonged periods of time, and in vitro stimulation of NK T cells derived from livers of chronically infected mice, compared to uninfected mice, results in increased gamma interferon and IL-4 secretion. Moreover, in NK T-cell-deficient mice the chronic-phase antibody response to a GPI-modified surface protein is decreased. These results indicate that, during the acute infection, NK T cells limit parasitemia and that, during the chronic phase, NK T cells augment the antibody response. Thus, during T. cruzi infection the quality of an individual's NK T-cell response can affect the level of parasitemia and parasite tissue burden, the intensity of the chronic inflammatory responses, and possibly the outcome of Chagas' disease.     

抗人重组肿瘤坏死因子β(rHTNF-β)单克隆抗体的研制及鉴定

肿瘤坏死因子β(TNF-β)是细胞毒性蛋白,与肿瘤坏死因子α(TNF-α)具有相似的生物学功能。为了进一步研究TNF-β的结构和功能,探讨TNF-β同TNF-α在生物学功能方面的相互关联,并且为纯化应用于临床的rHTNF-β提供技术手段。本实验应用脾内免疫杂交瘤技术制备了一株抗rHTNF-β单克隆抗体B5。Western blotting结果表明,B5特异地识别分子量为22kDa的TNF-β。B5和rHIL-1、rHIL-2、rHIL-6、rHIFN-γ、rHIFN-α、GM-CSF及E.Coli菌裂解液无交叉反应,B5培养上清与rHTNF-α有弱交叉,腹水在高浓度时有交叉反应。B5具有中和rHTNF-β细胞毒能力,亚类测定为小鼠IgG_1。     

Plasma Cells and their Precursors: II. Kinetics of B-Memory Cell Production in Rabbits

Rabbits were irradiated with 4.5 Gy in order to eliminate completely preexisting antibody-forming cell precursors. Sheep red blood cells were administered 24 h or 8 days after irradiation in order to induce the production of IgG B-memory AFCP. Resulting B-memory cells were triggered into antibody synthesis by a second dose of SRBC given 8 days after the challenge; the resulting IgG antibody clones were analyzed by isoelectric focusing. Memory IgG antibody clones were detectable from the third day after secondary immunization onward. It is concluded that antigen administered as early as 24 h after the irradiation induces B-memory cell production equally well as primary immunization 8 days after the irradiation. This B-memory cell production proceeds in the absence of detectable primary IgG antibody formation.Irradiated non-immunized rabbits showed spontaneous reappearance of IgG-AFCP with specificities to SRBC. In sharp contrast to the specifically induced production of B-memory IgG-AFCP mentioned above, this process took more than two months to reach potentialities comparable to those of «preexistent» AFCP present in normal, control rabbits.     

Alterations in the Distribution and Proliferative Responses of Rhesus Monkey Peripheral Blood and Spleen Cells During Malaria (Plasmodium knowlesi) Infection

Rhesus monkeys are used frequently as animal models in malaria research, but few studies have evaluated lymphocyte functions in these animals after experimental infections with the primate malarial parasite Plasmodium knowlesi. In this study, the distribution and mitogen responses of mononuclear cells in the peripheral blood and spleens of 16 P. knowlesi-infected rhesus monkeys were followed. All animals included in the study developed acute infections and were bled out with parasitemias of more than 50%. With progression of the infection, alterations in the peripheral blood mononuclear cells were observed, including decreases in the percentage of T cells (measured by E rosette formation) and the total numbers of E and EAC rosette-forming cells per cubic millimeter. In addition, peripheral blood mononuclear cells displayed reduced responses to mitogen stimulation with phytohemagglutinin, concanavalin A, and pokeweed mitogen. Peripheral blood mononuclear cells from infected animals showed similar reductions in mitogen responses when cultured in media containing 15% autologous pre- or postinfection plasma. The mitogen responses of spleen cells did not appear to be affected, but a significant reduction in the proportion of splenic T cells was observed. These lymphocyte changes in P. knowlesi-infected rhesus monkeys are similar to those reported for mice with acute rodent malaria and for humans with chronic Plasmodium falciparum infections.     

抗CCP抗体和RF联检在RA诊疗中的临床价值

目的：研究抗环瓜氨酸肽（anti-cyclic citrullinated peptide,Anti-CCP）（抗CCP抗体）和RF的检测在类风湿关节炎（RA）诊疗中的临床价值。方法：分别用酶联免疫吸附试验（ELISA）、BeckMan全自动蛋白分析仪同时检测早期RA组（病程〈1年）42例,RA组（病程〉1年）40例,非RA对照组40例患者血清抗CCP抗体和RF。结果：早期RA组、RA组的抗CCP抗体、RF阳性率显著高于非RA对照组（P〈0.05）RA组抗CCP水平显著高于早期RA组（P〈0.01）,两者RF无显著差别（P〉0.05）。RA组与早期RA组CCP抗体与RF二者无相关性。结论：联检抗CCP抗体、RF有助于类风湿的早期诊断和预测病情的进展。     

Glycolysis During Early Infection of Feline and Human Cells with Feline Leukemia Virus

A 30 to 40% increase in glucose uptake and lactic acid production was observed during a 48-h period immediately following inoculation of feline fibroblasts and HEp-2 cells with feline leukemia virus. There was no evidence of morphological alterations in either cell type. Immunofluorescent procedures to detect feline leukemia virus group-specific antigen revealed that the feline and HEp-2 cells were infected, whereas the antigen was not found in parallel cultures of uninoculated cells. Increased glycolysis depends upon the physiological state of the host cell and was not observed with infected stationary-phase cells.     

Platelet-Activating Factor Stimulates Interleukin-6 Production by Human Endothelial Cells and Synergizes with Tumor Necrosis Factor for Enhanced Production of Granulocyte-Macrophage Colony Stimulating Factor

The interaction between human endothelial cells (EC) and leukocytes during inflammation is in part mediated through the release of soluble factors. Since platelet-activating factor (PAF) is a potent mediator of inflammatory responses, we investigated the potential of PAF to modulate IL-6 and GM-CSF production by EC. Exposure of these cells to PAF resulted in a concentration-dependent increase in IL-6 production, with a maximum at 10^–10 M PAF. Sequential incubation of EC with PAF and TNF resulted in a synergistic increase of IL-6 production. This effect was specific for PAF since it was prevented by preincubation with the PAF receptor antagonist, WEB 2086. Northern blot analysis revealed enhanced IL-6 mRNA expression in PAF-treated EC. However, the synergy observed in protein synthesis between PAF and TNF was not reflected in IL-6 mRNA accumulation, suggesting a post-translational modulation. Pretreatment of EC with the protein synthesis inhibitor cycloheximide before their exposure to PAF resulted, after washout of the cycloheximide, in a markedly augmented production of IL-6, suggesting a synergy between augmented IL-6 mRNA accumulation by PAF and IL-6 mRNA superinduction by cycloheximide. GM-CSF production by EC was also stimulated by the combined effects of PAF and TNF, but PAF alone did not affect GM-CSF production. Taken together, our data suggest that PAF can stimulate EC to synthesize cytokines, including IL-6 and GM-CSF, which may contribute to local and, possibly, systemic responses during inflammation.