FEATURES OF THE IMMUNOGENETIC STRUCTURE OF PIGS OF DOMESTIC BREEDS, SUITABLE FOR XENOTRANSPLANTATION

The aim of the work was to determine the immunogenetic characteristics of pigs of two Ukrainian breeds by their adaptability, resistance, reproduction, genetic homogeneity, the presence of alleles that determine the potential suitability for use in xenotransplantation and other biomedical purposes. At present, there is no breed or specialized line of pigs in Ukraine to address the urgent problems of humane medicine. However, there is a favorable situation for the creation of such a breed or the withdrawal of a specialized line because of the demand of pharmacological concerns for model biological objects to study the mechanisms of action of modern medical preparations, to develop methods of conducting bloodless surgical operations, modeling of the clinic and epidemiology of infectious diseases of different diseases. immune response, etc. In this context, there was a compelling reason to preserve the indigenous breeds of pigs of Ukraine, especially Ukrainian meat and Мyrgorod. Standard methods of immunogenetic analysis of pigs were used. The erythrocyte antigens of 9 blood group systems were determined using specific immune sera and the involvement of a bank of immunodiagnostics meeting international requirements. Blood groups were determined by the reaction of agglutination, an indirect Coombs test, and a hemolytic test. Statistical processing of the research results was performed using mathematical statistics using the GenAlex-6.0 computer program. As a result of the immunogenetic analysis of Ukrainian beef and myrrh-pig breeds, it was determined that each of them is characterized by a specific immunogenic profile, which is related to both breed characteristics and differences, and methods of their breeding. The difference between the distribution of the overwhelming number of alleles of blood groups was statistically significant. Pigs of Ukrainian meat and Мyrgorod breeds had the largest differences in allele distribution by B, E, F, K, L blood group systems with the presence of the Ladhjk marker allele in the latter (p < 0.05). The specificity of immunogenic profiles was evaluated, depending on the history of creation and the direction of the animal's performance. Species of Myrhorod and Ukrainian meat breeds with the presence of genotypes A - / - and Ebdgkmp / bdgkmp were found to determine the suitability of animals for xenotransplantation. Of the 80 animals tested for immunogenetic markers, only 24 individuals met the established selection criteria. The range of determined indicators of actual homozygosity for selected genotype animals ranged from 55.56% to the maximum value – 88.89%, mainly in sows from the Smorodinа, Rusalkа, Sorokа and Soyka families. According to molecular genetic analysis, you have been informed that the established criteria of the biomedical model meet only 13 individuals of Ukrainian meat breed. Among animals with the highest index of actual homozygosity, representatives of Cererа and Cіlina were identified, with Celina 4092 being homozygous for all blood group systems studied. The selection of pigs for blood alleles, desirable for xenotransplantation, with their transfer to the homozygous state: A -/- and Ebdgkmp / bdgkmp will impair the adaptive, first of all, reproductive qualities of the animals and create significant problems for the cultivation of such individuals. The possibility of the occurrence of reproductive and resistance disorders of pigs in the selection of blood groups genotypes modeled for xenotransplantation and the search for ways to overcome them are being considered.

Immunogenetic analysis of brown Schwyz cattle lines

Animals of brown Schwyz breed from pedigree farms of the Smolensk region were differentiated by their belonging to lines and related groups. Identification by blood groups was carried out according to the generally accepted technique, and the genetic structure of the uterine population of each line was established using marker alleles of the EAB locus; 12 genealogical lines were identified. The number of alleles in individual lines ranged from 15 to 53, which indicates the necessary hereditary diversity for improving the breed. At the same time, the most common B2O3Y2A`2E`2G`P`Q`Y`, I1Y2E`2G`I`G``1, G3O1T1Y2E`3F`2G``, G1O` and b markers somewhat offset the differences between them. The coefficient of genetic similarity r between lines and related groups of animals ranges from 0.20 to 0.90. Interlinear differences in milk production of cows were established.

IMMUNOGENETIC PARAMETERS OF THE RED-AND-WHITE COWS IN RELATION TO THE LINEAR AFFILIATION

The paper explores the urgency of increasing efficiency of breeding Red-and-White dairy cattle that take the main part of the population in the Krasnoyarsk region (69.7%), the immunogenetic characteristics of animals that belong to Montwick Chiftein and Pabst Hoverner (n=52) lines. The research on the blood groups of cows were conducted in the laboratory of immunogenetic analysis of Krasnoyarskagroplem. The authors revealed the differences in the incidence of specific antigens in the cows of two lines. The article found out that antigens A’2, B2, E’3, O3, and T2 (P>0.95-0.99) were found to be more frequent in Montwick Chiftein line cows, and antigen P2 (P>0.95) wasfound in Pabst Governer line cows. It was also revealed that the most rare antigens in the Montwick Chifthein line are antigens A’, A’1, O2, O4, Y1, R1, X1, M and E’2, and antigens B’’, O4, U and U’’in the Pabst Governer line. Interestingly, the authors identified the antigens that were not present in the animals of both lines and in each line separately. The average index of antigen incidence in the cows of each line showed a relatively low value. Due to this the authors made a conclusion about a high degree of heterozygosity of the studied individuals. The index of genetic similarity among the animals of two lines demonstrates a high value of 0.863. The polymorphism of the antigens in the EAF-V locus was studied and a significant incidence predominance in cows of the F allele over the V allele was found out. The impact of genotype according to the EAF-V locus on dairy productivity and live weight of the cows was determined: in the Montwick Chifthein line, cows with the FF genotype (5986,60172,41 kg) had a higher yield, in the Pabst Governer line - with the FV genotype (5914,20208,19 kg). Animals of both lines that had genotype V/V had higher live weight parmeters than animals with genotypes F/V and F/F. The comparative analysis of the obtained results was carried out with the results of other Russian authors who conducted similar studies.

Distinct Immunogenetic Signatures in IgA Versus IgG Multiple Myeloma

Immunogenetic analysis of MM has proven instrumental in elucidating disease ontogeny e.g. by revealing the clonal relationship between switch variants expressed by the bone marrow plasma cells and myeloma progenitors in the marrow and blood; demonstrating the marked under-representation of the inherently autoreactive IGHV4-34 gene; and, identifying patterns of somatic hypermutation (SHM) indicative of post-germinal center derivation. Yet, limited information exists about the composition of the immunoglobulin (IG) gene repertoire in MM cases expressing different heavy chain isotype, in particular A versus G. This is relevant in light of studies showing an overall higher SHM impact in CD27+IgA+ compared to CD27+IgG+ normal memory B cells, perhaps reflecting a distinct location of the immune response, especially considering that IgA class switching mostly occurs in mucosa-associated lymphoid tissues. From a clinical perspective, it is also relevant to note that IgA patients exhibit a higher incidence of the t(4;14) translocation, shorter progression-free survival and worse median overall survival compared to IgG patients. Here, we explored potential differences in the immunoprofiles of IgA versus IgG MM focusing on IG gene repertoire and SHM characteristics. In total, 428 patients with a diagnosis of MM following the IMWG criteria from collaborating institutions in Greece, Italy and Spain (n=355) or retrieved from the LIGM-DB (n=73) were included in the study. Of these, 135 and 293 belonged to IgA and IgG MM groups, respectively. Amongst the evaluated productive IG rearrangements, IGHV3 subgroup genes predominated in both groups (IgA: 58.5%; IgG: 52.2%). However, at the individual gene level, major asymmetries were noted, since only 7 IGHV genes accounted for 41.6% of the IgA and 46.7% of the IgG cases, respectively. Of these, 3 genes were shared between IgA and IgG MM cases: IGHV3-30 (IgA: 11.9% - IgG: 13.3%), IGHV3-23 (IgA: 5.2% - IgG: 6.8%) and IGHV3-9 (IgA: 6.7% - IgG: 4.4%), whereas the remaining 4 of the 7 most frequent genes were specific for each group with significant (p<0.05) differences regarding the IGHV3-7 (5.2% in IgA versus 1.7% in IgG) and IGHV3-21 gene (0.7% in IgA 4.1% in IgG). IGHD3 predominated in both groups (IgA: 37% - IgG: 39.6%) followed by IGHD2 in IgG MM (18.4%) and IGHD6 in IgA MM (20.7%). IGHJ4 and IGHJ6 were the most frequent IGHJ genes with no significant differences in relative frequency. Searching for restricted IGHV-IGHJ combinations, we noted that the IGHV3-9 gene preferentially paired with the IGHJ6 gene in IgA MM versus the IGHJ4 gene in IgG MM (66.7% and 46.2% of all IGHV3-9 rearrangements, respectively). The median complementarity-determining region 3 (CDR3) length was identical in both IgA and IgG MM (15 amino acids, aa), yet differences were identified for specific CDR3 lengths as in the case of 19 aa, concerning 10.4% of all IgA versus 4.8% of all IgG rearrangements (p<0.05). Turning to SHM, the vast majority of rearrangements (IgA: 90.4%, IgG: 85%) were heavily mutated (IGHV germline identity (GI) <95%) with median GI of 91.8% for IgA and 92.2% for IgG. To study the topology of SHM, we compared the ratios of replacement (R) to silent (S) mutations in the framework (FR) and complementarity determining regions (CDRs) in cases expressing common frequent IGHV genes, namely IGHV3-23, IGHV3-30 and IGHV3-9 and identified distinct SHM patterns in all 3 instances: (i) IGHV3-23: the highest R/S ratios in IgA versus IgG MM were observed in FR2 (3.88) and CDR1 (3.9), respectively; (ii) IGHV3-30: overall "normal" SHM topology with higher R/S in CDRs rather than FRs, however, compared to IgG, IgA cases also showed a very high R/S in FR2 (5.3 versus 1.4); and, (iii) IGHV3-9: significantly (p<0.05) higher R/S ratios in CDR1 and CDR2 in IgG versus IgA cases (10.2 and 8.3 versus 1 and 2.9, respectively). Overall, in-depth immunogenetic analysis in the largest to-date series of IgA MM and IgG MM patients reveals differences regarding IGH gene repertoires, CDR3 characteristics and the topology of SHM. These findings suggest distinct antigen exposure histories and/or affinity maturation processes for IgA versus IgG MM, further highlighting the importance of microenvironmental stimuli in disease pathogenesis. Disclosures Terpos: Celgene: Honoraria; Novartis: Honoraria; Genesis: Consultancy, Honoraria, Other: Travel expenses; BMS: Consultancy, Honoraria; Amgen: Consultancy, Honoraria, Other: Travel expenses, Research Funding; Takeda: Consultancy, Honoraria; Janssen: Consultancy, Honoraria, Other: Travel expenses, Research Funding. Stamatopoulos:Gilead: Consultancy, Honoraria, Research Funding; Abbvie: Honoraria, Other: Travel expenses; Novartis: Honoraria, Research Funding; Janssen: Honoraria, Other: Travel expenses, Research Funding.