Detection of antibodies to HLA1 antigens by enzyme immunoassay

A modification of the MAILA (monoclonal antibody specific immobilization of lymphocyte antigens) method has been developed for the detection of antibodies to class 1 histocompatibility antigens. Russian biotin-treated monoclonal antibodies IKO-53 (Medbiospektr, Moscow) were used. In a complex with monoclonal antibodies, lymphocyte HLA antigen was found to retain its antigenic properties when stored for a long time. High specificity and sensitivity of the method were demonstrated. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 120, N ^o 9, pp. 315–317, September, 1995 Presented by V. I. Shumakov, Member of the Russian Academy of Medical Sciences     

Preparation of monoclonal antibodies to O-antigen (0–4.5) ofSalmonella typhimurium serogroup B

Sixteen clones secreting murine monoclonal antibodies of various specificities toSalmonella antigens are obtained. One of the clones is highly specific toSalmonella typhimurium (serogroup B) O-antigen in enzyme immunoassay. The resultant antibodies are IgG2a immunoglobulins with a “kappa”-type light chain. The connecting epitope includes abequose-mannose-rhamnose trisaccharide. Antibodies are bound toS. typhimurium whole cells and do not cross-react with representatives of theE. coli family. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 117, N ^o 6, pp. 630–632, June, 1994 Presented by I. P. Ashmarin, Member of the Russian Academy of Medical Sciences     

Specificity of monoclonal antibodies obtained by immunization of mice with trypsin-treated group a streptococcus culture

Hybridomas producing monoclonal antibodies intensively reacting with group A streptococcus antigens in enzyme immunoassay were obtained as a result of immunizing mice with pepsin-treated cultures of group A streptococcus. All antibodies were referred to class M immunoglobulins. The reactions of monoclonal antibodies were completely inhibited by the pepsin-treated culture of group A streptococcus. The degree of inhibition with A-polysaccharide was lower, being 17.5 to 50.0 in different monoclonal antibodies. All the monoclonal antibodies obtained cross-reacted with antigens of murine and human epithelial tissues of the thymus and skin. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 118, N ^o 11, pp. 492–495, November, 1994 Presented by A. G. Skavronskaya, Member of the Russian Academy of Medical Sciences     

Biodegradation and restitution of the intercellular matrix in preserved fragments of dense fibrous connective tissue after their grafting into the recipient

Results are presented from a study in which the structural arrangement of the intercellular matrix was examined in preserved connective tissue fragments (allografts) after their implantation into rabbits to repair posttraumatic space-occupying defects in the capsular-ligamentous complex of the knee joint. Stages of biodegradation and restitution undergone by the interstitial substance of connective tissues after the implantation of allografts are identified. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 121, N ^o 1, pp. 20–23, January, 1996 Presented by G. N. Kryzhanovskii, Member of the Russian Academy of Medical Sciences     

Rheumatoid factors of the blood and cerebrospinal fluid in patients with a complicated spinal injury

The levels of rheumatoid factors (RF) of IgM-RF and IgG-RF classes were measured by enzyme immunoassay (EIA) in the sera of patients with spinal injuries. Analysis showed that 46% of patients with spinal injuries were seropositive for IgG-RF-and 40% for IgM-RF; no IgG-RF was detected in the cerebrospinal fluid (CSF), whereas the IgM-RF content was 37.6 μg/ml. Results of RF measurements in the sera make it possible to qualify the systemic involvement of the body in the late period of a spinal injury as a rheumatic disease. The findings point to the presence of independent mechanisms of immune control in the CSF. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 118, N ^o 8, pp. 174–176, August, 1994     

Enzyme immunoassay system for detecting antibodies in complexes with HIV-1 antigens

An enzyme immunoassay is proposed for detection of antibodies in complexes with antigens. Specific anti-HIV antibodies were detected in sera which were considered HIV-negative. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 126, No. 8, pp. 189–192, August, 1998     

Role of various murine B-lymphocyte subpopulations in the immune response to T-independent antigens

Polyclonal antibodies to Lyb 5⁺ antigen of murine B lymphocytes are obtained and a methodological approach to the detection of cells carrying this antigen is developed with the aim of investigating the role of various subpopulations of mouse B lymphocytes in polyclonal activation induced by T-independent type 2 antigen. Hybridomas producing anti-Lyb 5.1 antibodies are obtained. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 119, N ^o 4, 402–405, April, 1995 Presented by B. F. Semenov, Member of the Russian Academy of Medical Sciences     

Synthesis and properties of the immunotoxin CD5-ricin

Synthesis and properties of an immunotoxin produced by conjugating ricin with a novel monoclonal antibody (IgG3 of the ICO-104 class) are described. Cytolytic activity of the synthesized immunotoxin, determined by two independent methods and expressed in LD₅₀, is 0.3–0.6×10⁻⁷ M. Its specificity for target cells containing the CD5 antigen is shown. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 119, N ^o 1, pp. 76–79, January, 1995 (Presented by Yu. A. Romanov, Member of the Russian Academy of Medical Sciences)     

Aspergillus fumigatus: Identification of 16, 18, and 45 kd antigens recognized by human IgG and IgE antibodies and murine monoclonal antibodies

The immunochemical properties of antigens produced by Aspergillus fumigatus were investigated with biochemical purification techniques in conjunction with the production of murine monoclonal antibodies (MAbs) and binding studies with human IgG and IgE antibodies. A. fumigatus antigens were partially purified by gel filtration and hydrophobic interaction chromatography on phenyl-Sepharose. Two fractions that eluted with either 2 mol/L or 0.15 mol/L of NaCl demonstrated strong binding to human IgG and IgE antibodies. Immunoprecipitation analysis with IgG antibodies from six patients with different Aspergillus-related diseases demonstrated that the 2M and 0.15M fractions contained major antigens of molecular weight 18 kd (Asp f I) and 45 kd, respectively. The ¹²⁵I-labeled 2M fraction was used to compare IgG antibodies to A. fumigatus in sera from 25 patients with Aspergillus-related diseases. IgG antibodies were significantly higher in patients with allergic bronchopulmonary aspergillosis (geometric mean, 437 U/ml) than in patients with asthma (geometric mean, 14 U/ml; p < 0.001), but undetectable (<5 U/ml) in 43148 control subjects. A good correlation was found between levels of IgG antibodies to the ¹²⁵I-labeled 0.15M fraction and the ¹²⁵I-labeled 2M fraction in sera from 106 patients with cystic fibrosis (r = 0.77; p < 0.001). Five murine IgG MAbs and two IgM MAbs were raised against the 2M fraction, and immunoprecipitation with the IgG MAb demonstrated two distinct antigens within the 2M fraction, Asp f I, and a 16 kd antigen. The results of a solid phase RIA with IgG MAb 4A6 demonstrated that ≈85% of A. fumigatus-allergic patients with allergic bonchopulmonary aspergillosis had IgE antibodies to Asp f 1. The three protein antigens defined in these studies are useful probes for investigating the immunopathogenesis of diseases associated with colonization by A. fumigatus.     

The new technology of hot start polymerase chain reaction

The use of monoclonal antibodies against Taq DNA polymerase in the polymerase chain reaction is proposed. These antibodies effectively inhibit polymerase activity at temperatures <70°C, activity being restored upon DNA melting in the very first stage of amplification. As in the “hot start” reaction, the addition of the antibodies to the incubation medium markedly improves the reaction yield. The method is particularly effective for the identification of a few copies of DNA. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 121, N ^o 2, pp. 238–240, February, 1996 Presented by V. N. Smirnov, Member of the Russian Academy of Medical Sciences     

Formation of colonies of clonogenic stromal precursors in bone marrow and splenic cell cultures in the presence of streptococcal antigens in the culture medium

The presence of streptococcal M protein and A polysaccharide in culture medium is shown to have an inhibitory effect on the growth of clonogenic stromal precursors in cultures of healthy murine bone marrow and of healthy guinea pig bone marrow and spleen. The efficacy of colony formation dropped 1.5- to 2-fold in the presence of antigens in a concentration of 25 μg/ml in the medium. The inhibitory effect was absent if antigens were added to adhesive cell cultures. The addition of antigens to cultures originating from animals immunized with streptococcus resulted in inhibition of the efficacy of colony formation in complete cultures and in cultures of adhesive cells. The presence of streptococcal antigens in guinea pig stromal fibroblast cultures of different strains did not affect their growth or colony formation. These data indicate that the effects of streptococcal antigens appear to be aimed at the stromal cells not directly, but rather via another cellular category in the bone marrow and splenic cell cultures, probably lymphocytes. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 118, N ^o 11, pp. 489–492, November, 1994     

Preparation and characterization of murine monoclonal antibodies toSalmonella typhimurium K-antigen

Monoclonal antibodies toS. typhimurium K-antigens representing IgG1 immunoglobulins with a “kappa”-type light chain are prepared and characterized. They are highly active in indirect enzyme immunoassay with purified K-antigens and whole fixedS. typhimurium cells and virtually do not cross-react with O-antigen of the same bacterial species or withE. coli antigenic structures. The kinetics of binding and ability to agglutinate whole bacterial cells are studied and epitope analysis is carried out, which shows that antigenic determinants ofS. typhimurium K-antigen qualitatively differ from those of O-antigen of the same bacterium. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 117, N ^o 6, pp. 633–635, June, 1994 Presented by I. P. Ashmarin, Member of the Russian Academy of Medical Sciences     

Autoantibodies to platelets in children with thrombocytopenia

Antiplatelet antibodies were titered in children and adults with idiopathic thrombocytopenic purpura. The levels of platelet-associated IgG were elevated in 100% of the examined children and 86% of the adults, their mean titers being at least 10 times higher than in the control and differing little within the groups. Serum antibodies reacting with normal donor platelets were detected in 43% of adults and in only 18% of children. Improvement of the clinical picture and increase of the platelet count in these patients according to different protocols always correlated with a drop in the level of antiplatelet antibodies. The results point to the immune nature of idiopathic thrombocytopenic purpura in children. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 120, N ^o 12, pp. 636–639, December, 1995 Presented by V. N. Smirnov, Member of the Russian Academy of Medical Sciences     

Localization and fate of thymic epitheliocytes of endodermal origin synthesizing cytokeratin 18

A reaction in the cytoplasm and processes of some cells of the epithelial reticulum is revealed by indirect immunofluorescence with monoclonal antibodies to cytokeratin 18 in the cortical zone of human thymus. In the medullary zone the reaction is observed in spherical elements similar in shape and size to intestinal goblet cells. Translated fromByulleten' Eksperimental'noi Biologii i Medsitsiny, Vol. 117, N ^o 2, pp. 188–190, February, 1994 Presented by V. I. Shumakov, Member of the Russian Academy of Medical Sciences     

The sera of patients with acquired immunodeficiency syndrome contain specific antibodies in a latent state

Latent antibodies reacting with antigens of human immunodeficiency virus are detected in patients with acquired immunodeficiency syndrome. Antibodies are detected by ion-exchange chromatography on QAE-Sephadex. A relationship is noted between the disease stage and the level of latent antibodies. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny Vol. 121, N ^o 3, pp. 315–316, March, 1996 Presented by A. F. Tsyb, Member of the Russian Academy of Medical Sciences     

A recombinant enolase from Anisakis simplex is differentially recognized in natural human and mouse experimental infections

A 1,963-bp cDNA was isolated from an Anisakis simplex cDNA library by immunoscreening with a hyperimmune rabbit serum raised against a crude extract of A. simplex L3 larvae. The open reading frame encodes a putative protein of 436 amino acid residues, which exhibits high similarity (70–80%) to enolase molecules from various other organisms, including helminth parasites. After subcloning and expression of the A. simplex cDNA in PGEX-4T-3, the resulting glutathione S-transferase fusion protein, purified by glutathione-Sepharose-4B chromatography, showed functional enolase activity. The immunogenicity of the recombinant A. simplex enolase was analyzed by immunoblotting using sera obtained from (a) mice immunized with crude extracts (CE) of A. simplex, or other nematode species, (b) mice immunized with excretory–secretory (ES) antigens from A. simplex, or (c) mice infected with L3 larvae by the intraperitoneal route. In addition, we used ELISA, to investigate the presence of IgG1 and IgE antibodies against this molecule in sera from patients infected with A. simplex. Mouse sera obtained after infection with L3 or raised against CE antigens, but not sera raised against ES antigens, showed strong reactivity with the recombinant A. simplex enolase. We also obtained good reactivity in Western blotting with sera from mice immunized with CE antigens from Ascaris suum and Toxocara canis, but not with sera from mice immunized with CE antigens from Trichuris muris, Trichinella spiralis or Hysterothylacium aduncum. In contrast to the experimental infections/immunizations in mice, we were unable to detect anti-enolase IgE antibodies in sera from human patients infected with A.simplex (15 sera), and the levels of anti-enolase IgG1 antibodies in these sera were low and apparently nonspecific. These results seem to indicate that, during natural infection in humans, A. simplex larvae do not offer sufficient antigenic stimulus to induce anti-enolase antibodies.Note: Nucleotide sequence data reported in this paper were submitted to the GenBank, EMBL and DDBJ databases under the accession number AJ496792.     

Dissemination of human solid tumor cells in the bloodstream of immunodeficient mice

Seven human solid tumor cell lines transplanted into hereditarily immunodeficient mice (nude and beige/nude) were typed for tumor-associated surface antigens and glycoconjugates using fluorescent conjugates of 7 monoclonal antibodies, 5 lectins, and 2 ligands. With this set of 14 selected tumor markers, peripheral blood samples from mice bearing the respective tumors were examined by flow cytofluorimetry for the presence of tumor cells disseminated in their circulation. Tumor cells were detected in the blood of mice carrying a uterine tumor, indicating that the metastatic process can be followed intravitally in immunodeficient animals bearing human solid tumors. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 120, N ^o 12, pp. 615–618, December, 1995     

Effect of neonatal blood serum on the early stages of mouse embryogenesisin vitro

The effect of blood sera from newborns delivered after complicated pregnancy and abnormal labor (CPAL) on 4-cell mouse embryos is studied. The embryos develop normally in the control sera, 78–100% of them reach the blastocyst stage, the mean number of blastomeres per embryo varies from 68 to 88 cells, and the mortality rate is 3–7%. The CPAL sera have an almost 100% embryolethal effect. The development of mouse embryos is markedly delayed. The most potent pathological effect is elicited by the sera of newborns whose mothers suffered from hypoxia. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 118, N ^o 7, pp. 6–7, July, 1994 Presented by A. A. Totolyan, Member of the Russian Academy of Medical Sciences     

Inhibition of platelet aggregation by monoclonal antibodies against glycoprotein IIb–IIIa complex

Monoclonal antibodies CRC64 are obtained against Ca²⁺-dependent glycoprotein IIb–IIIa complex of the platelet membrane which possess the ability to inhibit completely fibrinogen-dependent platelet aggregation. CRC64 is directed against the epitope formed by the glycoprotein IIb–IIIa complex and does not interact with proteins isolated after platelets are treated with ethylenediamine tetraacetate. Complete, reproducible blockade of platelet aggregation caused by 5 μM adenosine diphosphate is noted in an MCA concentration of 3 μg/ml, while in the case of a stronger inductor, namely 1 U/ml thrombin, platelet aggregation is inhibited in a concentration of 5 μg/ml. F(ab′)₂ fragments are also able to inhibit platelet aggregation completely and are usually effective in concentrations lower than native monoclonal antibodies. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 118, N ^o 10, pp. 402–405, October, 1994 Presented by V. N. Smirnov, Member of the Russian Academy of Medical Sciences     

Immunochemical analysis of common antigenic determinants in insulin and apoprotein B Molecules

A study is performed of the mechanism underlying stress diabetes, which develops in human beings and animals under stress. Dot-immunoanalysis shows the presence of common antigenic determinants in insulin, apoprotein B, and apoprotein B-containing low density and very low density lipoproteins isolated from human and rat serum. Electrophoresis, immunoelectroblotting, and immunoenzyme analysis reveal 5–6 peptides belonging to apoB, which specifically react with anti-insulin and anti-very low density lipoprotein antibodies. Insulinlike immunoreactivity is also identified in human serum supernatant obtained after precipitation of the total low density and very low density lipoprotein fraction and after removal all lipoproteins from it. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 118, N ^o 9, pp. 258–261, September, 1994