Paul-Bunnell antigen in lymphoma and leukemia spleens.

Lymphoid cells obtained from spleens of patients with lymphomas or leukemias were studied for the presence of heterophile (Paul-Bunnell (P-B)) antigen. A mixed agglutination (MA) test was established utilizing monolayers of cells attached to poly-L-lysine-coated wells of plastic U plates. After incubation of the monolayers with infectious mononeucleosis (IM) sera, indicator cells, sheep, or trypsinized bovine erythrocytes were added. The results were assessed according to sedimentation patterns of the indicator cells on the monolayers. Positive MA reactions were shown to be due to specific binding of P-B antibodies to the corresponding antigens on the spleen cells. Positive results were obtained with 15 of 37 spleens from patients with Hodgkin's disease, 5 of 8 lymphoma spleens, 4 of 15 chronic myelocytic leukemia spleens and 2 of 4 chronic lymphocytic leukemia spleens. Only 2 of 25 spleens from patients with various other diseases and 1 of 26 apparently normal thymus specimens gave positive results. This study confirmed demonstration of P-B antigen in lymphoma and leukemia by means of absorption experiments, which was reported previously.     

Hanganutziu-Deicher antigen and antibody in pathologic sera and tissues.

Heterophile, Hanganutziu-Deicher (H-D) antigen was studied in pathologic sera by means of inhibition of agglutination of bovine erythrocytes by H-D antibodies. H-D antigen was demonstrated in 38% of random cancer sera, 25% of lymphoma or leukemia sera, 25% of leprosy sera, 8% of infectious mononucleosis sera, 6% of rheumatoid arthritis sera, and 27% of synovial fluids of rheumatoid arthritis patients. None of 134 normal human sera gave positive results. Some of the inhibition-positive cancer sera formed precipitation lines with H-D antibody-containing sera. Over 50% of various extracts of cancer tissues as well as spleens of lymphoma or leukemia patients were shown to contain H-D antigen by means of the inhibition test.     

Comparison of the radioimmunological and immunoenzymatic methods in detection of HBsAg]

We have compared the solid-phase radioimmunoassay(SPRIA) with a solid-phase enzyme-immunoassay (EIA) in the detection of hepatitis B surface antigen (HBsAg). 708 sera from blood donors and 500 sera from patients with various diseases (acute and chronic hepatitis, chronic renal failure in hemodialytic treatment) were tested for HBsAg with both methods. 208 sera (17,2%) were found to be positive in SPRIA and 209 sera (17,3%) in EIA. Two HBsAg positive sera were tested in dilution series with both methods, too. The results show that the sensitivity and specificity of the EIA compare very favourably with those of the SPRIA.     

Antibodies to cytokeratin-8 in patients with different lung pathology

Antibodies to cytokeratin-8 were detected by enzyme immunoassay (EIA) in sera of 135 patients with cryptogenic fibrosing alveiolitis, different rheumatic diseases, sarcoidosis and exogenous allergic alveolitis, 109 patients with inflammatory lung diseases and 74 donors of the Moscow Blood Transfusion Station. The results revealed that The frequency of positive EIA reactions among the donors was 7%, while in the group of patients with rheumatic diseases--from 5.9% (scleroderma) to 42.9% (fibrosing alveolitis). Positive reactions also occurred in patients with exogenous allergic alveolitis and sarcoidosis. In the group of patients with chronic inflammatory lung diseases, i.e. in pathologies of non-autoimmune origin, positive reactions occurred in 13.3-33.3% of cases. To improve diagnostics and to disclose the mechanisms of pathogenesis, more detailed study of anticytokeratin antibodies in cases of interstitial lesions and chronic inflammatory lung diseases are necessary.     

Immunoblotting reactivity of human sera from various sources against purified epstein-barr virus

The immunoblotting technique was used to analyse polypeptides of purified Epstein-Barr virus reacting with antibodies present in sera from clinically healthy individuals, from patients with infectious mononucleosis (IM) or AIDS, and from renal transplant recipients with molecular sizes in the range of 40–290 kDa were detected.The 47- and 160-kDa nucleocapsid polypeptides, as well as the 72-, 74-, 140-, 220- and 290-kDa membrane polypeptides were the major viral proteins detected in the sera. Sera from clinically healthy individuals contained antibodies directed against all EBV membrane and nucleocapsid antigens. Sera from renal transplant recipients, from patients with IM and from patients with AIDS failed to react with certain nucleocapsid and membrane antigens; in particular, sera from AIDS patients and renal transplant recipients did not react with the 220-kDa polypeptide, one of the major membrane antigens, while sera from subjects with IM and from healthy individuals did.A high proportion of sera from patients with IM (38% vs 5% of clinically healthy individuals and 0–5% of the AIDS patients and renal transplant recipients) reacted with a 42-kDa polypeptide, suggesting its possible role in acute EBV infection.     

Determination of Mycoplasma pneumoniae antigens and antibodies by the immunoenzyme method in patients with respiratory tract diseases

The optimum conditions for carrying out the enzyme immunoassay (EIA) with a view to determine M. pneumoniae antigen and antibodies to them in the sera of patients with different respiratory diseases were established. The use of the specially modified EIA technique made it possible to reveal that patients with tuberculosis and chronic pneumonia showed similar occurrence of M. pneumoniae (35.7% and 35.0% of cases, respectively), while in patients with pulmonary sarcoidosis M. pneumoniae occurred in 27.2% of cases. At the same time the occurrence of antibodies in patients with chronic pneumonia and sarcoidosis was more than three times greater than in tuberculosis patients.     

Comparative evaluation of three recombinant antigen-based enzyme immunoassays for detection of IgM and IgG antibodies to human parvovirus B19

Background: Diagnosis of acute and past infection with parvovirus B19 is based on detection of IgM and IgG antibodies.Objectives: To evaluate two commercial recombinant antigen-based enzyme immunoassay (EIA) test kits for detection of IgM and IgG antibodies to parvovirus B19 and to compare the commercial EIAs to in-house EIA test procedures.Study design: A panel of 121 sera was used to compare the three IgM EIAs. The panel included 84 sera submitted for parvovirus B19 testing and 37 sera that were IgM positive for other viral pathogens. The same serum panel plus an additional 14 sera submitted for B19 testing was used to compare the three IgG EIAs. The commercial EIAs were performed according to manufacturers' instructions. Using the in-house EIA test procedures as the reference, sensitivity and specificity for each of the commercial EIAs was determined.Results: The commercial B19 IgM EIAs showed agreements of 95.0 and 93.4% to the in-house IgM EIA. Compared to the in-house B19 IgM EIA, the commercial B19 IgM EIAs, were 97.4 and 97.5% sensitive, respectively. Specificities were 93.5 and 91.4%, respectively. Sensitivities for the commercial IgG EIAs, compared to in-house IgG EIA, were 88.0 and 85.2%, respectively, and specificities were 94.1 and 98.0%.Conclusion: We found that the commercial parvovirus B19 IgM and IgG EIAs are comparable to standard in-house EIAs and are suitable for testing for B19 antibodies in human sera.     

不同戊肝抗原检测抗-HEV IgM反应性研究

目的比较不同戊肝抗原检测抗-HEVIgM反应性。方法用HEVE30、E42、E33合成肽和HEVORF-2重组抗原建立酶免疫试验（EIA）检测肝病患者和健康人群中抗-HEVIgM。结果60份抗-HEV阳性血清中,用E30、E42、E33及重组抗原包被检测抗-HEVIgM,阳性率分别为76.6%,26.6%,18.3%,66.7%。用E30抗原进一步检测戊肝急性期及恢复期血清,抗HEVIgM阳性率为90%及3.3%。结论以HEVE30为抗原的EIA特异性强、灵敏度高,是戊型肝炎早期诊断实用可靠的方法。     

Use of solid-phase immunoenzyme analysis for the serodiagnosis of pseudotuberculosis

The diagnostic test system based on the solid-phase enzyme immunoassay (EIA) for the detection of antibodies to Yersinia pseudotuberculosis in the sera of patients with the use of Soviet-made preparations and reagents has been developed. The test has been performed in microchambers for immunological reactions, thus making it possible to decrease the consumption of reagents 10-20 times in comparison with the traditional technique with the use of plates. The results of the titration of 42 sera in EIA and in the passive hemagglutination test (PHAT) are indicative of the presence of positive correlation (r = 0.78; p less than 0.05) between antibody titers in EIA and PHAT. A fourfold or greater increase in antibody titers has been determined by means of EIA in 80% of cases and with the use of PHAT in 55% of cases. The minimum diagnostic titer yielded by EIA has been determined: 1:256.     

不同戊肝抗原检测抗-HEVIgM反应性研究

目的　比较不同戊肝抗原检测抗 -HEVIgM反应性。方法用HEVE30、E42、E33合成肽和HEVORF 2重组抗原建立酶免疫试验 (EIA)检测肝病患者和健康人群中抗 HEVIgM。结果 6 0份抗 HEV阳性血清中 ,用E30、E42、E33及重组抗原包被检测抗 HEVIgM ,阳性率分别为 76 .6 % ,2 6 .6 % ,18.3 % ,6 6 .7%。用E30抗原进一步检测戊肝急性期及恢复期血清 ,抗HEVIgM阳性率为 90 %及 3 .3 %。结论以HEVE30为抗原的EIA特异性强、灵敏度高 ,是戊型肝炎早期诊断实用可靠的方法。     

Possible diagnosis of Pseudomonas aeruginosa infection by an immunoenzyme method using pyoimmunogen as the antigen

The possibility of detecting P. aeruginosa antibodies in patients by means of indirect solid-phase EIA techniques is shown. This assay is carried out with the use of reagents produced in the USSR: polystyrene assay plates manufactured by the Lenigrad Medpolymer Works are used as carriers, P. aeruginosa vaccine (pyoimmunogen) obtained under semi-industrial conditions at the Mechnikov Central Research Institute for Vaccines and Sera is used as antigenic complex and the commercial preparation produced by the Gamaleia Research Institute of Epidemiology and Microbiology serves as conjugate. The studies have revealed that in 95% of cases the level of antibodies in the sera of patients with acute destructive pneumonia accompanied by pleural empyema, abscesses of internal organs and acute hematogenic osteomyelitis is essentially higher than the level of "normal" antibodies in healthy donors from whom biologically confirmed P. aeruginosa cultures can be isolated. In the groups of patients with similar nosological forms of diseases caused by other infective agents such difference in antibody titers is not detected. These results suggest that the detection of antibodies to P. aeruginosa in patients' sera by means of EIA can be used as an additional test for the diagnosis of P. aeruginosa infections.     

Viral antibodies in infectious mononucleosis

Abstract Patients with Epstein-Barr virus (EBV) infectious mononucleosis (IM) usually develop heterophilic antibodies and some autoantibodies. Antibodies to rubella, measles, adeno-, entero-, herpes simplex, cytomegalo- and varicella-zoster viruses were titrated in sera from IM patients and matched healthy controls using the complement fixation test (CFT) and the haemagglutination inhibition test. Except for herpes simplex virus and cytomegalovirus, the IM sera had significantly higher arithmetical and geometrical mean antibody titres and showed in most cases higher antibody prevalences in the CFT. The titre rise was most pronounced for rubella and measles antibodies, between 2- and 3-fold. There were no cases of very high titres occasionally seen in IM. The IM sera had higher total IgG serum levels than the controls, 17.27 g/1 and 11.8 g/1, respectively ( P < 0.001). The present data show that in addition to previously reported high levels of some autoantibodies and of heterophilic antibodies, there is a more general increase in IgG antibodies to commonly occurring viruses. This increase is most likely due to the polyclonal activation of B-lymphocytes following the binding of EBV to the complement receptor CR2 (CD21). When due consideration is given to the possible occasional occurrence of a false positive rubella IgM test, the raised antibody-titres will most likely not interfere with routine diagnostics.     

Studies on precipitating and hemagglutinating antibodies in systemic lupus erythematosus

We studied the precipitating and hemagglutinating autoantibodies in the sera of patients with various connective tissue diseases in general and lupus in particular. Saline soluble extract of goat thymus had adequate antigenic materials as compared to other organs. Twenty per cent of patients with systemic lupus erythematosus were positive for precipitating autoantibodies by immunodiffusion and 44% by counterimmunoelectrophoresis. Normal human subjects, nonrheumatic disease patients and patients with rheumatoid arthritis and progressive systemic sclerosis were all negative. Forty seven per cent of positive systemic lupus erythematosus sera showed two precipitin systems. Enzyme sensitivities were used as the basis of identification of most of the antigenic specificities. Passive hemagglutination was carried out to identify antibodies to non-histone nuclear protein and nuclear ribonucleo-protein antigens. Thirty eight % of systemic lupus erythematosus patients were positive by this technique. Passive hemagglutination although a highly sensitive technique could not detect antibodies against antigenic systems other than non-histone nuclear protein and nuclear ribonucleoprotein.     

Antibodies to Yersinia enterocolitica serotype 3 in autoimmune thyroid diseases

The prevalence of increased titres of antibodies to Yersinia enterocolitica (serotype 3) has been studied in sera from patients with various thyroid diseases. In contrast to the low prevalences of the antibodies in healty subject (24.3%), titres (greater than 10) of anti-Yersinia enterocolitica (anti-Yersinia) were found more frequently in patients with thyroidal disorders, especially in Graves' disease (70.0%). Furthermore, high titres of the antibodies (greater than or equal to 160) were found only in patients with Graves' disease. There was no significant correlation between the titers of anti-Yersinia antibodies and those of anti-TSH receptor antibodies in sera from patients with Graves' disease. In seven individual samples of sera, the anti-Yersinia antibody titer was high before treatment, but the decrease in the anti-TSH receptor antibody titer following treatment was associated with a simultaneous decline in anti-Yersinia antibodies in all of them. A highly positive and significant correlation between the titers of anti-TSH receptor antibodies and anti-Yersinia antibodies was obtained in each of them. These findings could be merely a reflection of the measurement of the cross-reaction of anti-Yersinia antibodies with anti-TSH receptor antibodies but the possibility of an association between Yersinia infection and the production of anti-TSH receptor antibodies in at least some patients with Graves' disease cannot be ruled out.     

Elaboration of enzyme immunoassay based on recombinant antigens and intended for diagnostics of syphilis

Enzyme immunoassay (EIA) using recombinant antigens for the detection of Treponema pallidum-specific antibodies in sera of syphilis patients was developed. Four low-molar-mass Treponema antigens (Tp15, Tp17, TmpA, Tp47) were investigated; 17- and 47-kDa proteins were demonstrated as immunodominant as they permitted to obtain the most sensitive EIA. Using a mixture of these proteins a 3rd-generation-EIA kit Dia-Syph was constructed, its sensitivity being 99.4% during tests of 165 sera of syphilitic patients. No false result was obtained on the commercial panel PSS01 (BBI, USA). The specificity of the elaborated test system (99.7%) was determined on 295 sera.     

Interactions of sera from patients with autoimmune diseases with cDNA expressed fragment of topoisomerase I and monoclonal antibodies against enzyme

The interaction of sera from 34 patients with different autoimmune diseases with the expressed fusion protein cloned in lambda gt11 vector (topoisomerase I--beta galactosidase) and monoclonal antibodies against enzyme was studied. It was demonstrated that 100% of Scl cases possessed positive activity against fusion protein. It was shown that this test is not absolutely specific for Scl, i. e. 57.1% of Sle and 84.6% of RA demonstrated positive activity against "topoisomerase test". Autoimmune sera had the positive activity against monoclonal antibodies for topoisomerase I. This activity was shown to be due to the presence of antiidiotypic antibodies against topoisomerase in the autoimmune sera.     

The detection of antibodies to recombinant interferon alfa-2a in human serum

Three different procedures have been used for detecting antibodies to Roferon-A (recombinant human interferon alfa-2a, rHuIFN alpha-2a) in the serum of patients who received this interferon as part of ongoing clinical trials: an antiviral neutralization bioassay (ANB), the standard method recommended by the World Health Organization (WHO), and the more recently developed radioimmunoassay (RIA) and enzymeimmunoassay (EIA). Although the three tests are based on different principles, the correlation among them was excellent. The assays show differences in sensitivities with the ANB being the least sensitive of the three. The EIA equals the RIA in sensitivity, reproducibility, accuracy and labor and provides the advantage of safety and convenience in the use of non-radioactive materials. Therefore, the EIA has been selected as the most suitable assay for initial screening of the sera of patients receiving Roferon-A for the presence of antibodies to this interferon. EIA positive sera are then tested in the ANB to determine whether or not neutralizing activities are present.     

The use of commercial immunoenzyme kits for the diagnosis of toxoplasmosis

The results of comparative study of sera obtained from donors and from several groups of patients with suspected toxoplasmosis are presented. The study has been carried out with the use of commercial enzyme immunoassay (EIA) kits: Toxoplasma gondii IgG EIA and Toxoplasma gondii IgM EIA manufactured by Labsystems (Finland), Sevatest ELISA IgG Toxo Micro I manufactured by Sevac (Czechoslovakia). Statistical processing of the results has confirmed the identity of these kits. The necessity of using evaluation criteria (the separation point, the scale for the interpretation of results) when working with the Sevac kits is emphasized. Comparative evaluation of antibody profiles in the sera under test suggests that the titer less than 1:1600 should be regarded as the separation point for these kits. IgM antibodies to T. gondii have been found only in 22% of patients with high titers of IgG antibodies.     

清洁级实验动物四种病毒抗体玻片酶免疫检测试剂盒的研制

本文报道对清洁级实验动物应排除的四种病毒（淋巴细胞脉络丛脑膜炎病毒、小鼠脱脚病病毒、鼠肝炎病毒和仙台病毒）抗体玻片酶免疫（ＥＩＡ）检测试剂盒的研制。四种病毒感染的细胞和对照细胞经冷丙酮固定于载玻片上制成特异性抗原和对照抗原,此四种病毒的抗血清各１０份和ＳＰＦ小鼠血清２０份分别与四种病毒的特异性抗原和对照抗原进行ＥＩＡ交叉试验,结果显示,抗原只与其相应抗血清发生特异性显色反应,与非特异性小鼠血清和ＳＰＦ小鼠血清不显色。与ＨＩ或ＥＬＩＳＡ方法比较,通过对１１２份普通小鼠血清进行测试,结果表明,ＥＩＡ对仙台病毒抗体的检出率（１９．６％）显著高于（＜０．００５）ＨＩ（６．３％）,对小鼠脱脚病病毒抗体的检出率（２３．３％）与ＨＩ（２１．４％）无显著性差异（Ｐ＞０．０５）。ＥＩＡ对淋巴细胞脉络丛脑膜炎病毒和鼠肝炎病毒抗体的检出率分别为１．８％和７１．２％,ＥＬＩＳＡ对两种病毒抗体的检出率分别为１．８％和６７．６％,两种方法对两种病毒抗体的检出率无显著性差异（Ｐ＞０．０５）。重复性试验表明两批四种病毒抗体试剂盒对１０８份小鼠血清两次测定的符合率为９６～１００％。四种病毒的ＥＩＡ抗原在－１８℃保存１２个月或在２－８℃保存３     

Antibodies to saline extractable tissue antigen in sera of patients with renal diseases

Multiple serum samples originating from 110 renal allograft recipients were examined against saline extract of normal human kidney by means of double diffusion gel precipitation. Eleven recipients were found to be positive; 99 of 106 sera from these patients were positive. Pretransplantation sera were available from 7 of these recipients and 6 patients were found "positive." The precipitation reaction was composed of one line. Identity reactions were formed between the lines produced by sera from all patients except 1. Sera of patients from end-stage renal disease produced similar reaction; however, only 3 of 234 sera from patients with nonrenal diseases precipitated the kidney extract. None of 154 normal sera were positive. Several positive sera also were positive in complement fixation tests with human kidney extract. Evidence was presented that the antibodies under study combined with a nonorgan-specific but species-restricted tissue antigen. The hypothesis was advanced that these antibodies are autoantibodies formed in response to a sequestered antigen released as a result of tissue damage. Apparently, the antigen is released frequently in immunogenic form from injury to kidney but infrequently from injury to other organs.