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AB Abdul-Karim AD Barnes IS Bevan AJ Howie D Adu HF Ross C Raykundalia D Catty 《Canadian Metallurgical Quarterly》1995,3(3):192-202
Several studies have addressed the possible importance of anti-epithelial cell antibodies in kidney transplantation using the A549 cell line as an in vitro model. In this paper we report our results using for the first time an enzyme-linked immunosorbent assay (ELISA) to detect the anti-A549 cell antibodies. Sera from 129 kidney transplant patients were tested for IgM anti-epithelial cell antibodies directed against the A549 cell line prior to transplantation; only three sera were positive (2.3%). 101 of these patients were then followed-up post-transplantation; sera were collected routinely at 2, 6 and 12 weeks and at the time of rejection episodes. All samples were also tested for cytomegalovirus (CMV) IgM antibodies. Sixteen patients developed anti-A549 IgM antibodies, and there was no correlation with acute graft rejection. Anti-epithelial antibodies showed no binding to sections of normal kidney or biopsies of rejected kidneys. Eleven patients were positive for anti-CMV IgM antibodies. In nine cases both IgM anti-A549 and IgM anti-CMV antibodies were found, which was a highly significant association (p < 0.001). Analysis of A549 cellular proteins by immunoblotting gave evidence for the presence of CMV polypeptides in the cell lysate. Electron-microscopic examination of A549 cell preparations revealed intracellular particles which were compatible in size with CMV. Polymerase chain reaction analysis confirmed the presence of a specific CMV DNA sequence in A549 cells of several batches from different sources. Our data strongly suggest that the A549 cell line used in several published reports is infected with CMV and that in the majority of cases the anti-A549 'anti-epithelial' antibodies found in renal transplant patients are anti-CMV antibodies. 相似文献
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B Benyahia R Liblau H Merle-Béral JM Tourani J Dalmau JY Delattre 《Canadian Metallurgical Quarterly》1999,45(2):162-167
The authors investigated the association of caffeinated coffee, decaffeinated coffee, and tea with myocardial infarction in a study of 340 cases and age-, sex-, and community-matched controls. The odds ratio for drinking > or = 4 cups/day of caffeinated coffee versus drinking < or = 1 cup/week was 0.84 (95% confidence interval (CI) 0.49-1.42) after adjustment for coronary risk factors (1 cup = 237 ml). The odds ratio for drinking > 1 cup/day of decaffeinated coffee versus nondrinkers was 1.25 (95% CI 0.76-2.04). For tea, the odds ratio for drinking > or = 1 cup/day versus nondrinkers was 0.56 (95% CI 0.35-0.90). In these data, only tea was associated with a lower risk of myocardial infarction. 相似文献
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Rickettsiae are gram-negative, obligate intracellular bacteria which have historically been divided into three groups: the typhus group, the scrub typhus group, and the spotted fever group (SFG). Recently, several new SFG rickettsiae have been characterized, and most of these species are associated with ticks and have, as yet, no known pathogenicity toward humans. Rickettsia massiliae, which is widely distributed in Europe and Africa, is one such rickettsia. In order to investigate the antigenic relationships between R. massiliae and other rickettsial species and to develop a more convenient methodology for identifying R. massiliae, we produced monoclonal antibodies against the type strain (Mtu1T) of R. massiliae by fusing immunized splenocytes with SP2/0-Ag14 myeloma cells. A panel of 16 representatives were selected from the 163 positive hybridomas identified on initial screening, and their secreted monoclonal antibodies were further characterized. The reactivities of these 16 monoclonal antibodies with a large panel of rickettsial species were assessed by the microimmunofluorescence assay. All species of the SFG rickettsiae reacted with the monoclonal antibodies directed against epitopes on lipopolysaccharide, which is the common antigen among the SFG rickettsiae. Some closely related species of the SFG, such as Bar29, "R. aeschlimanni," and R. rhipicephali, showed strong cross-reactivities with the monoclonal antibodies directed against epitopes on the two major high-molecular-mass heat-labile proteins (106 and 120 kDa). In addition, species-specific monoclonal antibodies demonstrated that R. massiliae is antigenically different from other rickettsial species. Moreover, these species-specific monoclonal antibodies were successfully used for identifying R. massiliae in the ticks collected from southern France, and are therefore potentially useful tools in the identification and investigation of R. massiliae in ticks in large-scale field work. 相似文献
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Five monoclonal anti-mouse-blastocyst IgG antibodies were raised by intrasplenic immunization of three mice with adhesive-stage mouse blastocysts. Each mouse received a total of 60-70 blastocysts which were either nitrocellulose-immobilized or living but irradiated. Tests for pre-implantation stage-specificity showed that the antibodies differed in specificity. None were specific for surface epitopes. One antibody recognized epitopes only on blastocysts. Other antibodies were able to discriminate between unfertilized and fertilized oocytes, uncompacted and compacted morulae, or delayed and adhesive blastocysts. By applying reduced SDS-PAGE and Western blotting to blastocysts the blastocyst-specific antibody was seen to be bound to a peptide of M(r) 34. 相似文献
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S Sato M Fujimoto H Ihn K Kikuchi K Takehara 《Canadian Metallurgical Quarterly》1994,130(10):1273-1277
BACKGROUND AND DESIGN: Recently, we detected antihistone antibodies (AHAs) in patients with localized scleroderma. However, the exact antigen specificity of AHAs in this disease is still unknown. Therefore, we determined the reactivity of AHAs with five individual histones and the correlation of AHAs with rheumatoid factor in localized scleroderma by means of enzyme-linked immunosorbent assay. Twenty patients with localized scleroderma who had IgG and/or IgM AHAs, as determined by enzyme-linked immunosorbent assay, were examined. These patients were classified into the following three subgroups: patients with generalized morphea (n = 11), patients with linear scleroderma (n = 6), and patients with morphea (n = 3). RESULTS: In generalized morphea, IgG AHAs strongly reacted with histones H1, H2A, and H2B; and IgM AHAs strongly reacted with H1 and H2B, as determined by means of enzyme-linked immunosorbent assay. The pattern of reactivity in linear scleroderma and morphea was similar to that in generalized morphea. A homogeneous immunofluorescent pattern on HEp-2 cells, which was produced by localized scleroderma sera, was completely abolished by absorption with total histones. By employing a latex agglutination test, IgM rheumatoid factor was detected in 60% of the 20 patients with localized scleroderma and at a frequency of 82% in those with generalized morphea. However, an absorption test of rheumatoid factor activity with human IgG revealed no cross-reactivity of AHAs with rheumatoid factor. CONCLUSIONS: Our data suggest that AHAs in localized scleroderma are directed against native chromatin, since H1, H2A, and H2B occupy a relatively exposed portion of chromatin. 相似文献
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OBJECTIVES: The aim of the study was to determine the prevalence and clinical significance of antibodies to individual histone components in systemic sclerosis (SSc). METHODS: Serum samples from patients with limited cutaneous SSc (lSSc; n = 42) and diffuse cutaneous SSc (dSSc; n = 28) were examined for IgG and/or IgM antibodies to individual histone components and complexes by enzyme linked immunosorbent assay (ELISA). RESULTS: The level of IgG antibody to total histones was significantly higher in lSSc and dSSc than in normal controls. The level of IgM antibody to total histones was significantly higher in lSSc, but not in dSSc, than in normal controls. IgG antibody to total histones tended to be increased in dSSc when compared with that in lSSc. On the other hand, IgM antibody to total histones tended to be increased in lSSc when compared with that in dSSc. Although SSc showed various antihistone specificities, H2B, H2A-H2B, (H2A-H2B)-dsDNA were main antigens recognised by IgG antibodies in both lSSc and dSSc. Although IgM antibodies to H2B and H2A-H2B were also detected in both lSSc and dSSc, serum samples from lSSc patients exhibited highest IgM reactivity with H1. CONCLUSION: SSc may be included among conditions in which heterogeneous antihistone antibodies are produced. IgM antibodies to the most accessible histone H1 may be related to mild clinical features (lSSc) and IgG antibodies to the inner core molecules of native histone such as H2B or complexes including H2B may be associated with severe clinical features (dSSc) in Ssc. 相似文献
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A questionnaire was sent to all Human Fertilization and Embryology Authority-registered reproductive medicine centres throughout the UK to survey their policy for the diagnosis and management of antisperm antibodies. Forty-eight responses were received from the 74 units that use husbands' spermatozoa for treatments (65%). Most centres use at least one test to detect antibodies, although a minority perform no tests on the basis that their clinical practice would be unaltered if antibodies were present. Positive tests are classed as clinically significant at levels varying from > or = 10% to > or = 50% for direct sperm binding tests (mixed antiglobulin reaction, immunobead test), and ranging from any positive reaction to > or = 1:32 for the microtitre tests (gelatin and tray agglutination tests, microimmobilization test). Strategies for managing affected patients include no intervention, artificial insemination and intrauterine insemination (IUI) using spermatozoa prepared by various techniques, in-vitro fertilization (IVF) with or without increased insemination concentration, and intracytoplasmic sperm injection. Criteria for the latter are diverse, some centres managing all antibody-positive patients this way, while others resort to it only in severe cases or after other treatments have failed. Half of the respondents occasionally or regularly employ steroids, either alone or in conjunction with IUI or IVF. Overall, it appears that much confusion exists as to how best to manage couples presenting with antibody-related infertility. 相似文献
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Fine specificity of the myelin-reactive T cell repertoire: implications for TCR antagonism in autoimmunity 总被引:1,自引:0,他引:1
SM Anderton SP Manickasingham C Burkhart TA Luckcuck SJ Holland AG Lamont DC Wraith 《Canadian Metallurgical Quarterly》1998,161(7):3357-3364
The use of altered peptide ligands (APL) to modulate T cell responses has been suggested as a means of treating T cell-mediated autoimmune disorders. We have assessed the therapeutic potential of TCR antagonist peptides in autoimmunity using murine experimental autoimmune encephalomyelitis (EAE) as a model. The Tg4 transgenic mouse expresses an MHC class II-restricted TCR specific for the immunodominant encephalitogenic epitope of myelin basic protein, Ac1-9 (acetylated N-terminal nonamer). We have used T cell lines derived from Tg4 mice to define the TCR contact residues within Ac1-9. APL with appropriate substitutions at the primary TCR contact residue were effective antagonists of Tg4 T cells. These antagonist APL, however, were found to induce EAE in susceptible, nontransgenic strains of mice. Underlying this, the Ac1-9-specific T cell repertoire of normal mice, rather than reflecting the Tg4 phenotype, showed considerable diversity in fine specificity and was able to respond to the Tg4 antagonist APL. Defining antagonist APL in vitro using T cell clones, therefore, was not a reliable approach for the identification of APL with EAE-suppressing potential in vivo. Our findings highlight the complexities of the autoreactive T cell repertoire and have major implications for the use of APL in autoimmune diseases. 相似文献
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The photoadduct between native DNA and 8-methoxypsoralen was characterized on the basis of UV and fluorescence characteristics, Tm, nuclease S 1 digestibility and hydroxyapatite column chromatography. Approximately 80% of photoadduct has at least one diadduct (crosslink) per DNA molecule and only thymine was modified to the extent of 69%. The photo-crosslink was highly immunogenic inducing antibodies having apparent association constant of 1.3 x 10(-9)M. The immune IgG was highly specific towards nucleic acid-furocoumarin crosslink site without appreciable recognition of B-conformation or nucleic acid bases. The antibodies could be used as probe to detect and quantitate nucleic acid modified with furocoumarins in health and disease. 相似文献
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Tartrate-resistant acid phosphatase (TRAP) is expressed abundantly by osteoclasts and is required for bone resorption. This enzyme is emerging as an important biomarker in bone pathology, both for histochemical identification of osteoclasts and as a serum marker of osteoclast activity and increased bone turnover. Rat and mouse models are becoming popular systems for studying osteoclast development, bone physiology and morphogenesis, and bone diseases such as osteoporosis. We have developed two unique antibodies to human TRAP purified from hairy cell leukemia spleen. Both antibodies (9C5 and 14G6) are suitable for immunohistochemistry of osteoclasts and macrophages. Only one (14G6) is capable of immunoprecipitating active TRAP from human cell lysates. Antibody 9C5 reacts with a denatured epitope of TRAP while antibody 14G6 probably reacts with a native, conformational determinant. The high degree of homology among TRAPs of various species predicts that these antibodies should be suitable for work in experimental animals as well as humans. Immunohistochemical staining, electrophoretic analyses, immunoprecipitation and immunoblotting assays of human rat and mouse TRAP were carried out to test the validity of these antibodies as cell markers in rodents. Both antibodies were suitable for immunohistochemistry in all species. Antibody 9C5 was suitable for immunoblotting of denatured TRAP of all species tested. Antibody 14G6 reacted with the native TRAP of humans only and failed to immunoprecipitate mouse or rat TRAP activity. Although TRAP is a phylogenetically conserved protein, subtle, species-specific determinants exist. Care should be exercised when anti-TRAP antibodies are used for immunoassay in experimental animals. 相似文献
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Solway Kenneth S.; Huntley Debra K.; Stedman James M.; Laughlin Philip R.; Belar Cynthia D.; Flynn Michael F.; Carrington Christine H. 《Canadian Metallurgical Quarterly》1987,18(2):176
We obtained information about the characteristics of internship programs not accredited by the American Psychological Association (APA) and of their interns. Surveys were completed by directors of 74% of the programs and 51% of the interns in these programs. The most prominent reason why these programs did not seek accreditation was financial, though several of these programs had been in existence for several years, and 65% of them indicated that they would eventually seek accreditation. More than half of the interns in these programs were from APA-accredited graduate programs; their choice of internship was closely related to the location of the site. Interns did rate APA accreditation as important in ideal site selection, although four factors were rated even more important. Over 350 interns train in nonaccredited programs, which suggests their importance in the development of professional psychologists. Efforts being made by the Association of Psychology Internship Centers (APIC) to apply minimum criteria to these programs seem well warranted. (PsycINFO Database Record (c) 2010 APA, all rights reserved) 相似文献
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L Prokesová L Tucková B Cukrowska H Tlaskalová-Hogenová R Barot-Ciorbaru 《Canadian Metallurgical Quarterly》1996,18(11):661-668
Nocardia opaca, a Gram-positive bacterium, is a potent source of immunostimulatory substances. Screening of sera of adult human donors revealed that all sera tested contained antibodies reactive with isolated Nocardia fractions (Nocardia delipidated cell mitogen, NDCM; Nocardia lysozyme digest, NLD; Nocardia water-soluble mitogen, NWSM; and fraction B). The respective values of reciprocal titres for IgM and IgG were in the range of 100 to 12,800, and 10 to 320 for IgA antibody isotypes, when NLD or fraction B were used as antigens in enzyme-linked immunosorbent assay (ELISA) tests. The level of antibodies directed to NDCM, a potent polyclonal B cell activator, was found to be the lowest. In vitro spontaneous as well as NDCM-induced production of antibodies to NDCM by human peripheral blood lymphocytes involved mainly the IgM class. Western-blot analysis demonstrated that antibodies in normal human sera react with nocardial antigens of molecular mass approximately 60, 40, 20 and 15-10 kDa. The same antigens were also recognized by rabbit and mouse hyperimmune sera, also confirming the immundominancy of these nocardial antigens in other species. The presence of anti-nocardia antibodies in human sera and their production by both stimulated and non-stimulated lymphocytes points to the natural sensitization of humans either by ubiquitous no-cardial components or by cross-reactive bacterial or food antigens. 相似文献
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We determined islet cell cytoplasmic antibodies (ICA) using rat pancreatic sections as a test substrate substitutive for human pancreatic sections by indirect immunofluorescent technique. ICA were measured in sera from 58 patients with insulin-dependent diabetes mellitus (IDDM), 456 with non-insulin-dependent diabetes mellitus (NIDDM), 50 patients with autoimmune diseases, and 110 healthy controls. Seventeen of 58 patients with IDDM showed recent-onset (within 3 months). ICA were also measured in some samples using blood group O human pancreatic sections, and the ICA titers were compared with those measured using rat pancreatic sections. The prevalence of ICA was 55.2% (32/58) in patients with IDDM, 1.5% (7/456) in those with NIDDM, 0% (0/50) in those with autoimmune diseases, and 0.9% (1/110) in the healthy controls. Of the 17 recent-onset IDDM ICA were positive in 14 (82.3%). In comparative study of titers for ICA using rat pancreatic sections or human pancreatic sections, rat pancreatic sections yielded ICA titers as high as human pancreatic sections did. These results demonstrate that ICA assay using rat pancreatic sections was disease-specific, and that antigenicity of the substrate was favorable to ICA. Rat pancreas presents the advantage of greater availability, while providing an identical substrate for ICA. In conclusion, rat pancreatic sections are useful substrate for detecting ICA. 相似文献
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E Co?ffier M Girard F Barré-Sinoussi B Meignier E Muchmore PN Fultz C LeClerc 《Canadian Metallurgical Quarterly》1998,14(12):1023-1034
The fine specificity of the anti-V3 antibody responses induced in chimpanzees immunized by various human immunodeficiency type 1 (HIV-1) candidate vaccines and challenged by heterologous strains of HIV-1 was analyzed by enzyme-linked immunosorbent assay (ELISA) and Pepscan epitope mapping. Two chimpanzees immunized with the recombinant canarypox virus ALVAC-HIV (vCP125) expressing gp160MN and boosted with purified gp160MN/LAI alone, then with both immunogens in combination, were not protected against challenge with HIV-1 SF2. Their sera mainly recognized one epitope of the V3 loop, located in the NH2-terminal half. By contrast, immunization of two other chimpanzees with purified gp160MN/LAI and boosting with a synthetic V3MN peptide elicited a strong anti-V3 antibody response with a broader specificity directed against multiple epitopes all along the V3 loop. These chimpanzees were protected against infection by HIV-1 SF2. However, when these two chimpanzees were challenged later with a HIV-1 clade E strain virus, they became infected. We failed to detect any reactivity with the peptide of the ectodomain of gp41 of sera harvested after immunization with the various immunogens or after challenge with HIV-1 SF2 or HIV-1 90CR402. These results demonstrated that anti-V3 antibodies with a restricted fine specificity were induced in chimpanzees immunized with gp160 purified or expressed by recombinant canarypox confirming our previous results obtained in three different species (human, guinea pig and, macaque). In contrast, a boost with the V3 peptide broadened antibody responses, suggesting that the mode of presentation of the V3 loop to the immune system strongly influences the epitope specificity of the resulting antibody response. 相似文献
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Six groups of genetically mercury-susceptible female SJL/N (H-2s) mice were exposed to mercury vapor at a concentration of 0.3-1.0 mg Hg/m3 air for 0.5-19 hr/day 5 days a week for 10 weeks. The absorbed doses were calculated to be between 75 and 2365 micrograms Hg/week/kg body wt (micrograms Hg/week/kg). The correlation between the dose and the concentration of Hg in kidney, spleen, and thymus was highly significant (p < 0.0001; Spearman's rank correlation test). The lowest observed adverse effect level (LOAEL) for serum IgG antinucleolar antibodies (ANoA) was 170 micrograms Hg/week/kg, corresponding to a renal mercury concentration of 4.0 +/- 0.76 micrograms Hg/g wet wt. The correlation between the absorbed dose and the ANoA titer was highly significant (p < 0.0001; Spearman's rank correlation test), and all mice were ANoA-positive at a dose of 480 micrograms Hg/week/kg. High-titer ANoA targeted the nucleolar 34-kDa protein fibrillarin. The LOAEL for B-cell stimulation, measured as an increase in serum IgG2a and IgG1 concentrations, was 360 micrograms Hg/week/kg, but the increase was fivefold higher and also included IgE at a dose of 690 and 2365 micrograms Hg/week/kg. The serum Ig concentrations peaked after 2-4 weeks and then slowly declined but, except for IgE, remained significantly increased during the entire exposure time. Glomerular, mesangial IgG immune complex (IC) deposits, accompanied by systemic vessel wall IC deposits, were first detected at a dose of 480 micrograms Hg/week/kg. The mesangium also showed increased titers of IgM IC deposits and complement factor C3c. The correlation between the absorbed dose, and the individual titer of IgG, IgM, and C3c, was highly significant (p < 0.0001; Spearman's rank correlation test). In conclusion, mercury vapor efficiently induced an autoimmune syndrome in genetically susceptible mice, and the LOAEL for the adverse effects varied in the order ANoA < B-cell stimulation < IC deposits. Comparing the body burden of mercury in mice at the LOAEL for autoantibodies with the body burden in populations of occupationally exposed humans suggests that the safety margin may be narrow for genetically susceptible individuals. 相似文献
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PURPOSE: To compare the presence in levels of antiovarian antibodies (AOAb) in the pre- and postovulatory stage from serum of infertile patients undergoing intrauterine insemination (IUI) or in vitro fertilization (IVF) with outcome of the procedures. RESULTS: Serum from 36 women undergoing IUI, 36 women undergoing IVF and 25 fertile, healthy controls were assayed for the presence of AOAb by a commercially available ELISA kit. AOAb was positive in 59.7% of infertile women, while none of the fertile controls were positive for AOAb. The levels of these antibodies increased as the patient age and the number of treatment attempts increased. Though the presence of AOAb did not affect oocyte recovery rate, it resulted in decreased fertilization rate, cleavage rate, and pregnancy rate in infertile women. CONCLUSIONS: Our studies suggest that AOAb may be a cause of infertility and presence of these antibodies could have adverse effects on the outcome of assisted reproductive techniques. 相似文献