Opportunities for diagnostics of congenital and acquired immunological disorders in the Republic of Belarus

Abstract

IntroductionCongenital and acquired immunological children's disorders are a significant public healthcare problem, and it is undeniably important to improve the laboratory diagnostics of this pathology by creating new, effective methods for early detection of disorders involving immune mechanisms.One of the modern and promising methods for assessing lymphocyte proliferation is evaluation TREC and KREC, which are formed as a result of recombination of T- and B-cell receptor genes and are markers of lymphocyte neogenesis. Materials and MethodsWe have examined the DNA of patients with a genetically confirmed diagnosis of PID (n = 70) (SCID (n = 13), AT (n = 13), NBS (n = 15), diseases of immune dysregulation (n = 14), X-linked agammaglobulinemia (n = 15)) and after HSCT (n = 35). As a set for this research, the developed method of assessing the immunity general mechanisms by multiplex RQ PCR has been used to determine TREC and KREC. The method has been created in the State Institution “Belarusian Research Center for Pediatric Oncology, Hematology and Immunology” (Republic of Belarus). ResultsPatients with SCID, AT, NBS, diseases of immune dysregulation had TRECs values significantly reduced (p < 0.01) as to the reference values of healthy children. TRECs do not differ from healthy children (p > 0.05) for patients with X-linked agammaglobulemia. Patients with AT, NBS, diseases of immune dysregulation, X-linked agammaglobulinemia had KREC significantly lower than the reference values of healthy children (p < 0.01). For patients after HSCT, immune reconstitution of the T-cell immunity has been observed after six months after transplantation; from this period, the number of TREC-positive (naive) T-lymphocytes increased without a dynamic decline by 1st year after HSCT. Immune recovery of KREC has been observed after 3 months after transplantation and, as well as TREC, the number of KREC-positive (naive) B-lymphocytes increases by 1st year after HSCT. ConclusionBased on gained results, the developed TREC/KREC method with high sensitivity and specificity makes it possible to detect immunodeficiency for patients with suspected primary immunodeficiency. Measurement of TREC/KREC after HSCT can provide clinically relevant information that can be used to assess current state of patient in the process of reconstitution functional T- and B-cell immunity without using additional methods, such as flow cytometry.