Features of the distribution of the antigens of Kell, Duffy, Kidd, MNS, Lewis, Lutheran blood group systems in blood donors in the Kirov region.

The frequency of occurrence of antigens of the Kell (Kpa, Kpb), Kidd, Duffy, MNS and Lutheran systems in donors of the Kirov region corresponds to the distribution of antigens characteristic of white Europeans. Antigens K (Kell system) and Lea (Lewis system) are detected in the population of the region much less frequently, antigen Leb (Lewis system) - more often than in the population of Europe. The presence of a registry of donors typed according to a wide range of red blood antigens is a prerequisite for the immunohematological safety of blood transfusions.

Role of HLA genes in development of HLA-alloimmunization in blood recipients.

A genetically determined predisposition to the development of HLA-alloimmunization as a result of blood transfusions is associated with the presence of HLA-alleles DRB1*04, DQA1*03:01, DQA1*05:01 and HLA-haplotype DRB1*04-DQA1*03:01-DQB1*03:02 in the genotypes of recipients. The risk of antibody production is reduced in patients with HLA-alleles DRB1*16, DQA1*01:02, DQB1*05:02 and HLA-haplotype DRB1*16-DQA1*01:02-DQB1*05:02.

Description of two new alleles: HLA-B*50:79 and HLA-DRB1*04:332.

Two novel HLA alleles HLA-B*50:79 and -DRB1*04:332 have non-synonymous mutations in exon 4 and exon 3, respectively.

Characterization of the novel HLA-C*01:195 allele.

The new allele HLA-C*01:195 showed one nucleotide difference with HLA-C*01:02:01:01.

Two novel HLA alleles, HLA-DRB1*14:223 and HLA-DQB1*03:01:49, detected in a Buryat individual.

Two new alleles were characterized by next generation sequencing in a Buryat individual.

[Features of distribution HLAalleles and haplotypes in Kalmyks.]

Conducted high-resolution HLA-typing loci HLA-A, -B, -C, -DRB1 and -DQB1 by massively parallel sequencing of 150 potential donors of hematopoietic stem cells from the Republic of Kalmykia. In the studied population, four new alleles identified that not previously registered by the International Committee on the Nomenclature of Factors of the HLA-system of WHO. During the HLA-typing identified: 29 alleles at the HLA-A locus, 44 - at the HLA-B locus, 26 - at the HLA-C locus, 15 - at the DQB1 locus, 37 - at the HLA-DRB1 locus. The following alleles have a frequency of more than 10%: HLA-A*02:01 (11,7%), HLA-A*01:01 (11%), HLA-B*51:01 (10,3%), HLA-B*58:01 (10,3%), HLA-C*06:02 (17,7%), HLA-C*03:04 (10,3%), HLA-C*03:02 (10%), HLA-DQB1*03:01 (26,7%), HLA-DQB1*02:02 (10%), HLA-DRB1*07:01 (11,7%). The most common HLA-A-B-C-DQB1-DRB1 haplotype is A*02:05-B*50:01-C*06:02-DQB1*02:02-DRB1*07:01 (3,7%). Deviations from the Hardy - Weinberg equilibrium not identified.

[Expirience introduction of quantitative analysis of chimerism after allogenic stem cell transplantation by real-time PCR with InDel polymorphism.]

Using data obtained from domestic and foreign sources, we formed a set of primers and fluorogenic probes for analyzing twentysix specific sequence polymorphisms and one reference gene. In the course of evaluating the effectiveness of real-time PCR, using the example of one of the markers (S01a), we obtained the optimal amount of DNA per reaction (70 ng), providing a resolution of at least 0.1% of the method with the ability to estimate linear chimerism. Formed panel of primers for genetic polymorphisms - InDel has a high degree of informational content for donor-recipient pairs of Russia. From January 2018 to June 2019, a quantitative assessment of the level of linear (CD3 +, CD34 +) and general chimerism was carried out for 28 patients of the clinic of the Institution. Finally, we analyzed patients who received allografts and present 4 different clinical situations that illustrate the informativity level of this method.

[Design and verification of the reagent kit for high-productive HLA-typing of potential donors of hemopoietic stem cells.]

The work describes the development of a reagent kit for high-performance HLA-typing using the Illumina MiSeq System platform. The developed reagent kit contains all the necessary components for target enrichment of five HLA genes, preparation of libraries, and software for automatic data analysis. The reagent kit verified on a 93 DNA samples with known genotypes. During the verification, the sensitivity and specificity of the reagent kit for each of the HLA loci were determined - for A and B they were 1.0 and 1.0, respectively, for the C-1.0 and 0.99 locus, for the DRB1 locus 0.98 and 0.99, for the DQB1 locus 0.98 and 0.93.

[The genetic characteristics of Novosibirsk donors of hematopoietic stem cells.]

The HLA-typing was carried out concerning of 200 residents of Novosibirsk, potential donors of hematopoietic stem cells on loci (HLA)-A, -B, -C, -DRB1. The study detected in mentioned population two new alleles non-registered previously by the WHO International Committee on nomenclature of factors of HLA-system. The analysis of distribution rates of HLA-alleles and haplotype revealed 16 alleles alternatives of locus HLA-A, 24-HLA-B, 13-HLA-C, 13-HLA-DRB1. The rate of frequency more than 10% is intrinsic to following allele alternatives: HLA-A*02 (29.25%), 01 (14%), 03 (13.5%), 24 (10.75%), HLA-B*35 (12.25%), 07 (12%), HLA-C*07 (29.75%), 06 (13%), 04 (12.5%), 12 (11.5%), 03 (10.75%), HLA-DRB*13 (15.25%), 07 (13.75%), 01 (13%), 11 (12.75%), 15 (12.75%), 04 (10.5%). The application of software Arlequin v. 3.1 established 239 possible gaplotypes HLA-AB-C-DRB1. The gaplotypes A*01-B*08-C*07-DRB1*03, A*02-B*13-C*06-DRB1*07, A*03-B*35-C*04-DRB1*01 with rate of frequency 4.5; 2.75 and 2,75% correspondingly. The The distribution of alleles and analysis of galotypes permitted to compare analyzed population with other Russian populations.

Identification of a new HLA-B*27 allele, B*27:133, in a Russian individual.

The sequence of HLA-B*27:133 differs from HLA-B*27:05:02 by one nucleotide change within exon 3.

[The analysis of detection rate of ambiguity of loci under HLA-typing by SSO technology].

The article deals with the results of HLA-typing of 1829 patients on loci HLA-A, HLA-B, HLA-DRB1 using kits of LABType SSO reagents (One Lambada, USA). The indicators of each locus ambiguity are determined: locus HLA-A - 4.43%, locus HLA-B--4.43%, locus HLA-DRB1--0.16%. The list of "rare" alleles was applied to analyze 13 types of determined ambiguities on locus HLA-A, 27 types on locus HLA-B, 3 types on locus HLA-DRB1.

Identification of phosphorylase kinase as a novel therapeutic target through high-throughput screening for anti-angiogenesis compounds in zebrafish.

Angiogenesis is essential for development and tumor progression. With the aim of identifying new compound inhibitors of the angiogenesis process, we used an established enhanced green fluorescent protein-transgenic zebrafish line to develop an automated assay that enables high-throughput screening of compound libraries in a whole-organism setting. Using this system, we have identified novel kinase inhibitor compounds that show anti-angiogenic properties in both zebrafish in-vivo system and in human endothelial cell in-vitro angiogenesis models. Furthermore, we have determined the kinase target of these compounds and have identified and validated a previously uncharacterized involvement of phosphorylase kinase subunit G1 (PhKG1) in angiogenesis in vivo. In addition, we have found that PhKG1 is upregulated in human tumor samples and that aberrations in gene copy number of PhK subunits are a common feature of human tumors. Our results provide a novel insight into the angiogenesis process, as well as identify new potential targets for anti-angiogenic therapies.

[Experience in using the LABType SSO reagent kits in the practice of a HLA typing laboratory].

HLA was typed from HLA-A, HLA-B, HLA-DRB1 loci (to the second place) in 443 patients, by applying the LABType SSO reagent kits (One Lambda, USA). The findings were analyzed using the software Arlequin version 3.1. There were no deviations from the Hardy-Weinberg law. Overall, the authors identified 16, 27, and 13 allelic variants ofHLA-A, HLA-B, and HLA-DRB1 loci, respectively. There were also 4 most common haplotypes of HLA-A-B-DRBII: HLA-A *03-B*35-DRB1 *01 (4.29%), HLA-A *01-B*08-DRBl *03 (3.5%), HLA-A*03-B*07-DRBI *15 (3.37%), and HLA-A *02-B*07-DRBI *15 (2.93%).

[Identification of clinical isolates of Burkholderia cepacia and assessment of their pathogenicity in animal experiments].

In experiments on animals study of pathogenicity of 9 clinical strains of Burkholderia cepacia isolated from patients with chronic lung diseases was performed. Preliminary identification of studied strains by means of biochemical and genetic methods allowed to establish their belonging to B. cepacia species. It was determined that 6 of 9 strains are epidemiologically significant. Experiments showed that bacteria of studied strains are not able to cause infectious process in white mice and hamadryas baboons. Conclusion about appropriateness of development and use of other biological models was made.