[The subpopulation structure of human T-lymphocytes studied by using a monoclonal antibody (MCA) to CD27]. / Izuchenie subpopuliatsionnoi struktury T-limfotsitov cheloveka s pomoshch'iu monoklonal'nogo antitela (MKAT) CD27.

The obtained murine mAb LT27 (IgG2a) assigned to the cluster of differentiation CD27 was used to study the distribution of antigen CD27 among human lymphocytes scbpopulations in normal state and immunopathology. In normal donors the antigen CD27 was found to be expressed most frequently on CD4+ cells (90 +/- 8% of which coexpressed antigen CD27) and to the lesser extent on- CD8+ cells (only 77 +/- 28% of CD8+ cells carried antigen CD27). 79 +/- 12% of double negative lymphocytes (CD3+CD4-CD8-) expressed antigen CD27. In patients with hypogammaglobulinemia the proportion of CD4+CD27+/CD4+ and CD8+CD27+/CD8+ was significantly reduced to 80 +/- 11% (p < 0.01) and 45 +/- 19% (p < 0.001), respectively. The ratio CD4+CD27+/CD4+ varied insignificantly with the increase of CD4+ population, but the increase of the CD8+ population was accompanied by the definite tendency to a decrease of the ratio CD8+CD27+/CD8+. The distribution of CD27 antigen inside CD4+ and CD8+ subpopulations was found to be different from the distribution of CD29 and CD45RA antigens.

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.

[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.

[Cytometric method of determining the parameters of the interaction of synthetic immunostimulants with the lymphocytic surface]. / Tsitometricheskii metod oprdeleniia parametrov vazimodeistviia sinteticheskikh immunostimuliatorov s poverkhnost'iu limfotsitov.

The cytofluorimetric method was suggested for the determination of parameters of polymer adsorption on the cellular surface. Multiple equilibrium in the homogeneous cellular population with N independent binding sites and in the heterogeneous population of cells with the Poisson distribution of adsorption sites was analysed. A possibility was shown of cytofluorimetric drawing of the adsorption isotherm using mean fluorescence intensity of cells in the equilibrium suspension. The equilibrium association constant and the mean number of binding sites were estimated for adsorption of poly-L-lysin on the surface of human myelogenous leukemic cells (cellular line K-252).

Redistribution of subpopulations of peritoneal macrophages with Fc gamma receptors under the effect of an immunomodulator.

The heterogeneity of resident peritoneal macrophages and peritoneal macrophages was studied in different periods following oral administration of sodium nucleinate according to their ability to bind and phagocytize sheep erythrocytes opsonized by means of specific rabbit IgG. Using a mathematical method developed earlier, it has been possible to demonstrate that resident peritoneal macrophages can be divided into two subpopulations--actively and poorly binding macrophages but, after activation by sodium nucleinate--into three subpopulations. Fractionation of the macrophages according to their ability to adhesion within a temperature gradient has shown that the same peaks are traced in the fractions as in the overall pool of cells, but in different quantitative ratios. It has also been demonstrated that phagocytic activity is reduced in macrophages capable of adhesion to plastic at lower temperatures.

[Poisson equalization of multicomponent cytofluorograms]. / Puassonovskoe vyravnivanie mnogokomponentnykh tsitofluorogramm.

A method of the Poisson fitting of flow cytometry data is suggested. The estimation of distribution parameters was carried out by the steepest descent method. The phagocytosis of fluorescent latex particles by murine macrophages was used as an experimental model. The results obtained by the Poisson fitting of cytofluorograms were in good agreement with the visual estimation of macrophage distribution with regard to the amount of uptaken particles. A high value of correlation index (more than 0.99) showed the validity of the suggested fitting method. The applicability of the method to cytofluorimetric analysis of human lymphocytes was successfully demonstrated.

[Analysis of macrophage heterogeneity by their phagocytic activity]. / Analiz geterogennosti makrofagov po ikh fagotsitarnoi aktivnosti.

Heterogeneity of resident peritoneal macrophages was studied as revealed by their phagocytosis and binding of sheep erythrocytes opsonized by specific rabbit IgG. It has been shown that the macrophage heterogeneity is due to the presence of active and inactive cellular subpopulations. A mathematical model, based on the Poisson fitting of experimental histograms, was elaborated for the analysis of subpopulation composition of macrophage pool in the test of rosette formation and phagocytosis. The validity of the model was supported by physicochemical isolation of the macrophage subpopulation which was inactive in erythrocyte binding. The macrophage pool was separated into fractions by absorption--elution at different temperatures. Active and inactive macrophage subpopulations were found in all the fractions but in different ratios.

[Structural and functional analysis of the respiratory burst in the phagocytosing macrophage]. / Strukturno-funktsional'nyi analiz "vspyshki dykhaniia" u fagotsitiruiushchego makrofaga.

The interrelation between structural changes and oxygen consumption by the phagocyting macrophage was studied. The mean number of phagocyted particles was estimated by the method of stereological transformation. It is found that the uptake of yeast particles and CN- -nonsensitive oxygen consumption is related to the concentration of yeast cells in the incubation medium. A positive correlation was established between the oxygen consumption and the mean number of phagocyted particles. The results obtained may suggest that the "respiration burst" takes place in the contact area of the macrophage and the phagocyted material, and its extent probably depends on the surface of that contact area.

[Stereological analysis of phagocytosing cells]. / Stereologicheskii analiz fagotsitiruiushchikh kletok.

Various complexes of phagocytes with adsorbed (I) and entrapped (J) particles are formed during phagocytosis. Method of stereological reconstruction is proposed that allows to demonstrate the actual distribution of these complexes on the basis of morphometric analysis of their ultrathin sections. The principle of the method lies on the probability simulation of section distribution for a given distribution of complexes and on the solution of the reverse problem by stepwise determination of the relative quantity of each complex type (from the most complicated to the most simple one, when I = 0 and J = 0). The stereological analysis of phagocytosing murine peritoneal macrophages revealed an absolutely different and more adequate kinetical picture of phagocytosis, as compared to the morphometric data.

[Structural and functional study of acidification in the process of phagocytosis]. / Strukturno-funktsional'noe izuchenie zakisleniia v protsesse fagotsitoza.

Morphometry and cytofluorometry of the kinetics of phagocytosis were applied to study the quantitative mechanisms of acidification in the area of contact between the plasma membrane of macrophages and Candida albicans yeast cells conjugated with fluorescein isothiocyanate. It was found by stereological transformation of the morphometry data that the main part of macrophages phagocytose the limiting amount of particles by minute 5-10. A good agreement was established between the cytofluorometric histograms and the stereological data. This made it possible to evaluate, using the calibration curve, the pH on the surface of the phagocytosed material based on the fluorescence quenching. As an advantage of the suggested comprehensive approach to the study of acidification, the authors stress the possibility of making structural-functional analysis of the rearrangements occurring in the intact phagocytosing cell.