Two-colour flow cytometry study of lymphocyte subpopulations in patients with primary immunodeficiencies.

Immunofluorescent flow cytometric examination of one hundred and eighty-five children with different primary immunodeficiency syndromes and sixty-nine control patients revealed twenty-six cases with a bimodal distribution of antigens CD5 and CD7. Such abnormalities were most frequently found in patients with total antibody deficiency, namely those with common variable hypogammaglobulinaemia (10/24 patients) and congenital agammaglobulinaemia with lack of B cells (10/40), but were never seen in normal controls. Two-colour flow immunofluorescence demonstrated that antigen CD4 was expressed only on intensely fluorescent CD5+ cells, irrespective of the immunodeficiency state. Antigen CD4 was detected on cells with both high and low expression of antigen CD7, but a small percentage (2%-5%) of CD4+ lymphocytes did not belong to the CD7+ population. Antigen CD8 was found equally on intensely and weakly fluorescent CD5+ and CD7+ cells. In some immunodeficient patients suffering from ataxia-telangiectasia (12/36) and in some with Wiskott-Aldrich syndrome (2/6) there was a significant excess (greater than 20%) of CD7+ over CD5+ cells. In these patients a considerable number of the CD8+ cells were not part of the CD5+ population, but were always part of the CD7+ population. Cell populations with the phenotype CD5-, CD7+ consisted mainly of lymphocytes showing weak expression of antigen CD8.

[Study of human lymphocyte subpopulations using a panel of monoclonal antibodies]. / Issledovanie subpopuliatsionnogo sostava limfotsitov cheloveka s pomoshch'iu paneli monoklonal'nykh antitel.

Characteristics of newly produced monoclonal antibodies (MCA) against differentiation antigens of human T- and B-lymphocytes and monocytes have been presented. The features of new MCA were compared with those of MCA with predefined specificity. According to two-color immunofluorescence and radioimmunoprecipitation data some of the former MCA were assigned to established clusters and the other were considered to recognize previously noncharacterized antigens. Potential value of MCA against different antigens in immunological diagnosis has been discussed.

[The expression of an antigen interacting with monoclonal antibody IB4 on the surface membranes of normal human leukocytes and in lymphoproliferative diseases]. / Izuchenie ékspressii antigena, vzaimodeistvuiushchego s monoklonal'nym antitelom IB4, na poverkhnostnykh membranakh leikotsitov cheloveka v norme i pri limfoproliferativnykh zabolevaniiakh.

Monoclonal antibody (MAb) 1B4 of IgG3 isotype reacting with 55 +/- 10% peripheral blood lymphocytes was obtained. MAb 1B4 is bound with 100% of B and NK cells and 50-70% of T cells in blood. The most part of CD8+ cells (60-100%) and approximately 50% of CD4+ cells belong to 1B4+ population. T-ALL cells do not express 1B4 antigen. Analysis of reactivity of MAb 1B4 with transformed malignant cells has shown that 1B4 antigen appeared in the process of differentiation of B lymphocytes at the state of pre-B cells and disappeared with activation of B lymphocytes and transition to plasma cells.

[An evaluation of the fine structure of a human lymphocyte population based on the data of 2-color flow immunofluorimetry]. / Otsenka tonkoi struktury populiatsii limfotsitov cheloveka po dannym protochnoi dvukhtsvetnoi immunofluorimterii.

A set of 2n basis lymphocyte subpopulation, distinguishing by the presence of at least one antigen, may be composed for a given combination of n surface antigens. The relative amounts of basis subpopulations constitute a thin subpopulation structure of lymphocytes. General methods of lymphocyte phenotyping using one (or two) differentiation antigen give actually the estimation of a sum of basis subpopulations expressing a given antigen (or some combination of two antigens). The mathematical method was elaborated for reconstruction of the thin subpopulation structure by fitting the relative assemblage of biparametric cytofluorograms. The proposed approach was used for the subpopulation analysis of lymphocytes from patients with different forms of immunological deficiency.

Pancreatic islet cells in tissue culture: function and immunoreactivity with serum autoantibodies from diabetes type-1 patients and monoclonal antibodies ICA-1.

The purpose of this study was the generation of monoclonal antibodies (MoAB) ICA-1 to p64 antigen of pancreatic beta cells and the comparative analysis of MoAB and insulin-dependent diabetes mellitus (IDDM) patients' serum reactivity with cultivated islet cells. It was found that MoAB ICA-1 reacted with the surface of beta cells in suspension but with cytoplasm in frozen pancreatic sections. That MoAB ICA-1 and ICSA- or ICA-positive IDDM serum on insulin release in islet cell cultures produce similar effects was established. These results were obtained from in vivo injection of MoAB into rats and resulted in serum insulin decrease, increase of glucose concentration and morphological changes in pancreas corresponding to IDDM, which testified to the biological role of p64 antigen in pathogenesis of IDDM.

[Membrane markers of T- and B-lymphocytes in children with primary immunologic deficiency conditions]. / Membrannye markery T- i B-limfotsitov pri pervichnykh immuno- defitsitnykh sostoianiiakh.

Altogether 79 children with primary immunodeficiency conditions (PIC) and 21 children suffering from frequently occurring bacterial infections were examined for the level of rosette-forming cells and lymphocytes reacting positively with monoclonal antibodies to different phenotypic membranous markers (SD-antigens and others). The patients with combined forms of immunodeficiency, with T deficiency playing the key role manifested a significant lowering of the content of the subpopulations of SD3+, SD5+, SD7+ and SD4+ lymphocytes and the decrease of the SD4/SD8 ratio. The the reduction of the number of B lymphocytes (smIg + K1, SD19 +), the low levels of Ia+ -cells, and the decrease of the SD4/SD8 at the expense of the content of SD8+ -cells. In the patients with selective immunodeficiency and recurrent bacterial infections without verified PIC, these parameters did not undergo any essential changes. As compared with immunofluorescent assays, the measurement of E-RFL and M-RFL amounts was found to have insufficient information content for the diagnosis of PIC.

[Study of the characteristics of various hematologic diseases using monoclonal antibodies LT1, LT2 and LT7]. / Kharakteristika nekotorykh gematologicheskikh zabolevanii s pomoshch'iu monoklonal'nykh antitel LT-1, LT-2, LT-7.

Monoclonal antibodies (MCA) LT1, LT2 and LT7 were characterized. It was established that MCA LT1 and LT2 (IgG1 and IgM classes, respectively) interacted with antigen 67 kD, and MCA LT7 (IgGZa)--with antigen 40 kD. MCA LT1, LT2 and LT7 were shown to identify normal and neoplastic cells of T-lymphoid type. MCA LT1 and LT2 also reacted with B-lymphocytes of chronic lymphocytic leukemic patients. MCA LT1 and LT2 were referred to CD5 (Cluster of Differentiation) and MCA LT7--to CD7 by the molecular weight of an identified antigen and the spectrum of its expression. The importance of MCA studied in the diagnosis of subtypes of acute and chronic lymphocytic leukemia has been confirmed.

[A panel of monoclonal antibodies against human lymphocyte antigens]. / Panel' monoklonal'nykh antitel protiv antigenov limfotsitov cheloveka.

Monoclonal antibodies (McAb) LT1, LT2, LT4, LT6, LT7, LT8, LB21 and LDR to surface antigens of human lymphocytes have been obtained. Basing on the data concerning the distribution of McAb reactivity with cells from different organs and tissues, and with tumour cell lines, on experiments on the mutual blockage and comodulation of antigens, on two-colour immunofluorescence and determination of the molecular weight of the detected antigens a conclusion is made that the McAb obtained react with CD5, CD2, CD4, CD6, CD7, CD8, CD21 and HLA-DR antigens, respectively. McAb LT1 and Leu1, as well as LT2 and Leu5b in pairs blocked each other completely. The determinant recognized by McAb LT8 is more completely overlapped with the epitope, reacting with McAb OKT8. The McAb LT4 binding site is only partially overlapped with the McAb Leu3 site of recognition and practically is not bound to the epitope recognized by McAb OKT4.