The method of analysis of concentration of immunosuppressant of cyclosporine A in whole blood was developed. The highly effective liquid chromatography with mass spectrometric detection was applied using device of ionic trap type. The optimal conditions of analysis are established. The tryout of method was carried out using blood samples of healthy donors and patients underwent allotransplantation of organs. The comparison was made of the developed method with method of fluorescence polarized immunoassay Abbott TDX applied in clinical diagnostic. The higher selectivity of the proposed method to cyclosporine A as compared with Abbott TDX was established.

The method of analysis of concentration of immunosuppressant of cyclosporine A in whole blood was developed. The highly effective liquid chromatography with mass spectrometric detection was applied using device of "ionic trap" type. The optimal conditions of analysis are established. The tryout of method was carried out using blood samples of healthy donors and patients underwent allotransplantation of organs. The comparison was made of the developed method with method of fluorescence polarized immunoassay Abbott TDX applied in clinical diagnostic. The higher selectivity of the proposed method to cyclosporine A as compared with Abbott TDX was established.

It is known that lymph enters the heart through lymphatic vessels associated with large blood vessels and spreads from the endocardium to lymphangions of myocardium and epicardium. From the epicardium, where lymph plays an important role in homeostasis of pericardial fluid, it enters the sub-epicardium and leaves the heart through 2 extracardiac lymphatic vessels for regional lymph nodes. Endothelial cells of lymphatic and blood vessels share many common features, including surface expression of human leukocyte antigen (HLA) type I and II. This makes them a possible target for activated lymphocytes and anti-HLA antibodies after heart allotransplantation. This study used immunohistochemical and morphologic methods to examine lymphatic vessels in endomyocardial biopsy specimens after allotransplantation.

Urine sediment from 56 renal transplant recipients was tested cytologically. The recipients had 7 episodes of acute rejection (AR), 8 episodes of cyclosporin nephrotoxicity (CNT), acute tubular necrosis (ATN), combined complications, bacterial infection in 11, 6 and 16 cases, respectively. 12 patients had stable renal function. Cytospin preparations were made for Pappenheim staining. FACScan cytograms were obtained based on light scattering (cell size and granularity). Low cell count in patients with stable renal function and higher levels of mononuclear cells in all cases of a complicated postoperative period were seen. In acute tubular necrosis the sediment had a high scatter pattern, i.e. high density and large particles, while in cyclosporin nephrotoxicity renal epithelial cells had a low scatter pattern. Polymorphs were typical of the bacterial infection and acute tubular necrosis. Acute rejections were associated with elevated lymphocyte number. Microscopic examination revealed predominance of collect duct cells, lymphocytes with morphological signs of activation in acute rejection and proximal tubular cells in cyclosporin nephrotoxicity. Episodes of combined complications were difficult for diagnosis due to their mixed cellularity.

Endomyocardial diagnostic biopsies, recipient heart removed at operation, endomyocardial biopsies of allotransplants and postmortem material were studied using immunofluorescence to specify immunopathological process and to detect humoral rejection. Altogether 306 samples from 55 patients were studied. In the early postoperative period (one year) 8 out of 18 patients with heart transplants repeatedly showed immunopathologic picture of acute humoral (vascular) rejection which was characterized by a widespread immunoglobulin G and complement fixation in the capillary walls accompanied by enhanced capillary permeability and fibrin deposition in intestitial tissue. Such patients often had graft dysfunction. 1 to 5 years after transplantation in 24 out of 37 patients discrete focal immunoglobulin and complement fixation was observed as one of chronic rejection component.

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.

Reaggregation of pancreatic islet cells during their cultivation in stationary culture leads to formation of neoislets--compact insulin secreting clusters. By the means of indirect immunofluorescence it has been shown that neoislets have no leukocytes-passengers, common for normal islets and responsible for allograft rejection.

Immunofluorescence microscopy of endomyocardial biopsy specimens from heart allograft recipients identified immunopathologic changes in three of 17 patients. These changes included immunoglobulin G and complement C3 deposition in tissue structures such as capillary endothelium and basal membranes, cardiomyocyte sarcolemma, and interstitial tissue. Moreover, the immunopathologic changes could be correlated with acute cellular rejection episodes evidenced by endomyocardial biopsy criteria.

Myocardial tissues of patients with dilated cardiomyopathy were studied by immunofluorescence. While immunoglobulin A fixation was observed in myocardial capillary wall and cardiomyocyte sarcolemma in the majority of patients (11 of 12), immunoglobulin G and C3 complement component were a rare finding. In the vessel wall of heart allografts immunoglobulin A fixation occurred 3-6 days after transplantation. As a result of the intensive immunosuppressive therapy which was used after the operation immunoglobulin A disappeared from heart allografts within 4-5 weeks. Immunoglobulin A fixation in the heart of patients with dilated cardiomyopathy is attributed to the anti-tissue or antivirus antibodies and probably is involved in the development of this disease.

Peripheral blood mononuclear cells from Type I diabetic patients health donors or Wistar rats were cultured with rat islet cells for 18 h. Then TNF content of the medium and basal and stimulated insulin release were determined. Mononuclear cells from both healthy donors and diabetic patients could inhibit the insulin release with no correlation to TNF content. Addition of lipopolysaccharide resulted in a 5-7 times increase of TNF content of the medium followed by a more pronounced inhibition of insulin release. Rat mononuclear cells inhibited the beta-cell function almost completely and initially produced large amounts of TNF. The data indicate that inhibition of insulin release by blood mononuclear cells in vitro does not reflect anti beta-cell specific cellular immunity, involves cytokines and, probably depends upon the initial properties of the cells.

Purified rat islets were dissociated into single-cell suspension with an EDTA-Trypsin treatment. During a stationary culture in vitro the islet cells reassociated forming aggregates (neoislets). Electron microscopy revealed that the aggregates consisted mostly of beta-cells and not numerous alpha-cells. They showed a good insulin-secreting capacity and were able to increase insulin release in response to glucose plus theophylline. The lack of passenger leukocytes makes the neoislets particularly suited for experimental and clinical transplantation.

Morphofunctional alterations occurring in the colon wall indicate complex interactions of the morphological structures participating in the regulation of the intestinal function in salmonellosis. In the early period (24 hours) of maximum decrease in the mediator activity in the nerve structures the number of enterochromaffin cells and the granulation index increased, while in the period of maximum tension of nerve elements (7 days) a decrease in the enterochromaffin cells and granulation index was noted. At later periods (21 days) a relative morphofunctional stabilization of the mucous components and the intramural ganglia has been revealed.

It has been found that two active in neutral medium thiol proteinases from bovine spleen, cathepsin L and cathepsin H, bring about rapid and irreversible inactivation of alpha 1-proteinase inhibitor (alpha 1PI)--one of the major plasma inhibitors of serine proteinases. The activity of the enzymes studied did not change upon the interaction with alpha 1PI. With stoichiometric proteinase/inhibitor ratio, the inactivation of alpha 1PI under the effect of cathepsin L was instantaneous, while under the effect of cathepsin H it occurred within 30-60 min. The products of alpha 1PI inactivation had an inhibitory effect on the rate of its reaction with cathepsin L. alpha 1PI inactivation under the action of cathepsin L and cathepsin H was accompanied by the decrease in the molecular mass of the inhibitor from 54 kDA to 46 kDa. This was, probably, caused by the hydrolysis of the peptide bond formed by NH2 group of threonine. The 46 kDa fragment did not undergo further degradation. It did not bind to immobilized trypsin but retained antigenic properties. The results obtained show that the limited proteolysis is a mechanism of the inhibitor inactivation. It is suggested that under some conditions thiol proteinases, upon their release from the cell, participate in the control of effective alpha 1PI concentration.

A study was made of the sensitivity to prednisolone (PS) of peripheral blood mononuclear cells (MS) of patients with systemic lupus erythematosus (SLE) with renal lesion showing resistance to glucocorticoids (GC). It was shown that in most of the patients (60%) MC were poorly inhibited by PS whereas the rest of the patients (40%) demonstrated in vitro sensitivity compared to that of the donors and even higher in some cases. Thus, the group of SLE patients showing clinical resistance to GC action was inhomogeneous in sensitivity of their MC to PS; only some patients showed resistance associated with the properties of the MC system. Hemocarboperfusion was shown to produce a considerable effect on MC sensitivity to PS: higher sensitivity was noted in all the cases, it was particularly noticeable in the group of patients with low basal sensitivity to PS. An analysis of the data obtained led to a conclusion that hemocarboperfusion raised MC sensitivity to PS. This mechanism was shown to determine the efficacy of the method of therapy of resistance to GC.