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Biomedical subjects

S Kosinski

Publications and source records attributed to S Kosinski.

7 recordsLinked to original sources

Beta2-ADR haplotypes/polymorphisms associate with bronchodilator response and total IgE in grass allergy.

Association and linkage studies of beta2-adrenergic receptor (beta2-ADR) polymorphisms in relation to the expression of asthmatic phenotypes and immune regulatory mechanisms have shown inconsistent results. In order to analyse the relevance of particular combinations of single nucleotide polymorphisms (SNPs) or haplotypes of beta2-ADR gene to bronchial asthma, bronchodilator response and total immunoglobulin E (IgE) we determined by direct DNA sequencing five SNPs (in positions: -47, -20, 46, 79, 252) in a group of 180 Caucasian subjects (110 patients with grass allergy and 70 nonatopic controls). The eight different beta2-ADR haplotypes were identified, with three the most common of them representing 92% of the studied cohort. Significantly higher (pcor = 0.0045) bronchodilator response was observed in patients with homozygotic genotype 46A/A in comparison with respective homo- and hetero-zygotes. There was no significant difference in bronchodilator response when beta2-ADR haplotypes were analysed. Significantly higher (pcor = 0.0005) total IgE levels were found in patients with beta2-ADR haplotype -47T/-20T/46A/79C/252G and homozygotic carriers of 46A (pcor = 0.0015) and 79C (pcor = 0.003) genotypes. No significant associations were found in regards to asthmatic phenotype and atopy. These results indicate that depending on phenotype studied, either an individual beta2-ADR SNP or beta2-ADR haplotype might affect disease manifestation.

Adult↗

A supertypic HLA class II determinant shared by DR1 and DRw9, and crossreactive with DR2, defined by human monoclonal antibody.

A human monoclonal antibody Pez.2F5, produced by a lymphoblastoid cell line, has been established in vitro by Epstein-Barr virus (EBV) transformation of B lymphocytes isolated from the blood of a volunteer immunized with allogeneic peripheral blood leukocytes (PBLs). The antibody reacted with a new supertypic determinant expressed on all lymphoblastoid cell lines homozygous for HLA-DR1, -2, and -w9. The genetic linkage of the Pez.2F5 determinant to the HLA region was demonstrated by family segregation studies. Quantitative absorption studies indicated that DR2-positive cells required more Pez.2F5 antibody for lysis, and since their absorption capacity was significantly lower than that of DR1- or DRw9-positive cells, it is likely that the Pez.2F5 determinant of the DR2 haplotype is crossreactive but not identical with the determinant found on the latter haplotypes. In addition, on a test panel of HLA-typed B lymphocytes, Pez.2F5 showed perfect correlation with DR1 and DRw9, but reacted with only a fraction of DR2-positive cells. The Pez.2F5 determinant was found to be absent from resting T lymphocytes, but its expression could be identified on IL-2-dependent T-cell lines by cytotoxicity and flow cytofluorometric analysis. By sequential immunoprecipitation and SDS gel analysis of antigens of DR1 cells it was determined that the Pez.2F5 determinant is carried by HLA class II DR molecules. Thus, the Pez.2F5 is the first described human monoclonal antibody able to immunoprecipitate HLA class II-related molecules.

Antibodies, Monoclonal↗

A human monoclonal antibody against HLA-A25.

We are reporting the production and characterization of a human monoclonal antibody recognizing antigen HLA-25. The antibody was developed by a line transformed in vitro by the Epstein-Barr virus. The immune B lymphocytes for transformation were generated by planned immunization of a volunteer with repeated doses of allogenic peripheral blood lymphocytes of one donor over the course of 7 years. The antibody showed correlation with A25 antigen on a panel of 244 individuals tested by microcytotoxicity. The antibody showed neither cytotoxic reactivity nor CYNAP phenomenon with antigens of HLA-10 CREG.

Antibodies, Monoclonal↗

A new cloning method for antibody-forming lymphoblastoid cells. Increase in cloning efficiency by inclusion of human fibroblasts into semisolid agarose growth layer.

We report the development of a method for cloning human EBV-transformed cells which has greater efficiency than techniques used presently. In this new method lymphoblastoid cells are cultured in semisolid agarose in close physical association with human fibroblasts. The results indicate a 10-fold increase in the cloning efficiencies. The average cloning efficiency, depending on the age of cell lines, was from 1 to 14%, and colonies appeared 7-9 days sooner than in the traditional soft agarose method. The new method has allowed us to develop several stable lymphoblastoid cell lines producing antibody cytotoxic to human B lymphocytes. This method may make it more practical to obtain monoclonal human antibodies from lymphoblastoid cell lines which had previously been unstable due to heterogeneity.

Antibody-Producing Cells↗

A human monoclonal antibody to an HLA-DRw53 (MT3)-like epitope on class II antigens.

We report the production of a lymphoblastoid human B cell line, which secretes monoclonal antibody with HLA-DRw53-like specificity. The immune lymphocytes were obtained from a patient who had been undergoing immunotherapy at Memorial Hospital with allogeneic cultured melanoma cells. The antibody producing cell line was established by transformation of immune B cells with Epstein-Barr virus and stabilized by serial selection of antibody-positive cell clusters. The cell line (R39.14) has been producing a cytotoxic antibody of IgM, kappa type during nearly two years of continuous culture. The specificity pattern was determined on a panel of HLA-DR-typed lymphoblastoid cell lines, and resembled the HLA-DRw53 specificity closely. In studies of informative families the R39.14 specificity segregated with DRw53-bearing haplotypes, which included DR-4, 7 and DRw9 antigens. The R39.14 specificity was found to be expressed in similar quantities on all antibody positive homozygous cell lines tested. Blocking experiments with monoclonal class II specific antibodies revealed R39.14 to reside on a DR molecule in close association with the determinant detected by the 109d6 mouse monoclonal antibody.

Antibodies, Monoclonal↗