In practical terms, regardless of HLA compatibility level, whenever tissues are transplanted from one person to another it is essential to suppress the immune response of the recipient. A variety of methods are available however, the most frequently used ones have the disadvantage of being immunologicaly non specific. The consequence is a difficult balance between immunosuppression sufficient to prevent the tissue rejection and maintenance of immune system at the level of ability to adequately deal with an infection. The goal, not yet achieved, is to find a way of generating donor specific immunosuppression that leaves the immune machinery otherwise completely intact. The major approaches to immunosuppression are described below.

Increased level of cyclosporine in whole blood leads to different severe adverse reactions causing disturbance in function of kidney as well as liver, central nervous system, increased blood pressure, gingival hipertrophy. Low doses of cyclosporine lead to HVGR (host versus graft reaction). Cyclosporine level in whole blood does not depend upon dose only. So the level of cyclosporine has to be determined regularly to avoid either severe adverse reactions of the drug or HVGR. In the present study we analysed cyclosporine levels in 15 patients after kidney transplantation. Cyclosporine levels were determined by fluorescence polarization immuno assay, and the monoclonal whole blood was carried out by analizer ABBOT - TDX. In these patients the cyclosporine level determinants were performed six times during three months. In intervals of 15 days, at the exactly same time, we followed the parameters determining the kidney and liver functions : creatinin, ALT(Alanin aminotranssferasis), AST(Aspartat aminotranssferasis), serum concentrations of bilirubin, and ultrasound diagnostics of kidney and liver). The results of our study have shown that frequent monitoring of cyclosporine concentrations in whole blood in patients with transplanted kidney is extremely important It is the way how to maintain the cyclosporine levels in whole blood in the recommended interval between 100 and 300 ng/ml. By doing it the adverse drug reactions of cyclosporine in our study were reduced to a minimal level.

Molecular typization and subtypization of leukemia by RT-PCR, is important due to monitoring of minimal residual disease (MRD), during and after treatement of patients with acute leukemia In our experiments of PCR optimization, we used BIO-RAD procedure for multiplex RT-PCR screening and split-out PCR for leukemia subtypization. This method is adapted or optimized for Perkin Elmer Gene Amp 9600 thermal cycler and Qiagen HotStar Taq DNA polymera.se as well. According to manufacturer instructions, using of some other PCR engines require optimization of corresponding PCR parameters. We used Perkin Elmer 2400 thermal cycler and Hot Star Taq DNA polymera.se. Total RNA was extracted from whole blood specimens, obtained from Clinics of haematology-KCU Sarajevo, taken from patients with acute leukemia, transferred to cDNA, and amplificated in PE 2400. Detection of PCR products is performed by 1,5% agarose gel electrophoresis. Obtained optimal PCR parameters were: annealing temperature -65°C, number of cycles for both, first round PCR amplification and nested PCR amplification was 30, final concertation of MgCl in reaction mixture was 3,28 mM. Instead recommended amount of cDNA (5 µl) and Hot Star Taq DNA polymera.se (0,4 µl) we used amounts of 10 µl and 1 µl By using of this optimized PCR protocol we detected genetic aberration inv(16)(p 13; q22) and by using of split out PCR, subtype G(192 bp electrophoretic band)(CBFB / M YH11 fusion genes).