OBTAINING DONOR GAMMAGLOBULIN FROM MULTIPAROUS WOMEN, AND ITS EFFECT ON THE EXPRESSION OF HLA-G AND HLA-DR MOLECULES IN LYMPHOCYTES OF WOMEN WITH CHILDREN WITH SEPTAL CONGENITAL HEART DEFECTS

Abstract

The common link in the pathogenesis of reproductive losses and congenital heart disease (CHD) is associated with immune inflammation in the "mother-embryo" system, which affects the differentiation and proliferation of progenitor cells of the cardiovascular system. It is hypothesized that this link can be blocked by regulatory autoimmune and alloimmune antibodies to HLA-G and HLA-DR molecules. In addition, these antibodies may be present in sufficient amounts in donor immunoglobulins, especially those obtained from the blood of multiparous women. Based on this, the aim of the study was to obtain a purified gammaglobulin fraction from the blood of multiparous women and evaluate its functional activity in relation to HLA-DR and HLA-G molecules.

Results. Isolation of the gammaglobulin fraction (GGG) from the blood plasma of multiparous women wasperformed using affinity chromatography in several sessions. The purity of the resulting protein wasanalyzed by immunoelectrophoresis, electrophoretic separation of the protein fraction of blood serum and electrophoresis in 4.12% polyacrylamide gel with the addition of SDS (PAGE electrophoresis). PAAG electrophoresis showed that FHG did not differ from commercial therapeutic intravenous immunoglobulin (HIG).

An assessment of the functional activity of FHG in relation to HLA-DR and HLA-G molecules wasformed: the main group of women and their children with congenital heart disease (n=38) and the control group of women with their conditionally healthy children (n=21). To determine the specificity of FHG with respect to HLA-G, HLA-DR molecules, as well as to compare its effect with autologous and allogeneic sera and HIG, an immunological testing protocol using flow cytometry was developed. The protocol was developed on the basis of the methodological approach - "cross-match", as well as the patent of the Russian Federation "Method for determining antibodies to HLA-G". It was found that the blocking activity of female serum in relation to autogenous (intrinsic) and allogeneic (embryo/fetus/child) HLA-G and HLA-DR molecules determines the protective effect on the formation of congenital heart defects in the next generation. Donor human immunoglobulin has similar blocking effects to these molecules, possibly due to the presence of alloimmune antibodies to HLA classes I and II. The gammaglobulin fraction obtained from the donor blood of multiparous women has a more pronounced blocking effect on the expression of HLA-G and HLA-DR molecules. Thus, this immunobiological preparation can be a prototype of a therapeutic and prophylactic agent blocking the pathogenesis of congenital heart defects.