EFFECTS OF FEMALE BLOOD AUTOSERUM ON ALLOGENIC INTERACTIONS IN SHORT-TERM LYMPHOCYTE CULTURES OF PARENTS HAVING CHILDREN WITH CONOTRUNCAL HEART MALFORMATIONS

Highlights The findings of this original study ensure the detection of violations in the humoral regulation of the maternal immune interactions with semiallogeneic fetus, considered as a risk factor for developing sporadic conotruncal heart malformations in the next generation.
Aim To study the role of female autoserum blood in limiting allogeneic interactions in short-term lymphocyte cultures of parents having children with conotruncal heart malformations.
Methods 21 married couples (the study group) with children suffering from conotrucnal heart malformations (Tetralogy of Fallot) without chromosomal diseases were examined. The control group consisted of 21 families with three or more healthy children. The immune response in a mixed lymphocyte culture of parents was assessed by the increase in HLA-DR expression in the mixed culture with respect to spontaneous lymphocyte cultures. Primary staining of female and male lymphocytes with monoclonal antibodies to CD45, conjugated with various fluorescent dyes (PC-5 and PC-7), allowed assessing the immune response of female lymphocytes to male and vice versa.
Results The effects of female autoserum on the mixed lymphocyte culture of parents were assessed. The obtained results reported that the birth of children with conotruncal heart malformations is associated with the interfering effect of female autoserum on HLA-DR expression on subpopulations of female lymphocytes (CD3+, HLA-DR+) and the activating effect on subpopulations of female lymphocytes (CD3-, HLA-DR+). The observed role of female autoserum in the study group may be associated with the absence of HLA-DR-blocking autoantibodies and high synthesis of cytokines by T2 and T3 helper lymphocytes.
Conclusion The effects of female autoserum on allogeneic lymphocyte interactions of parents may be observed in short-term mixed lymphocyte cultures. The evaluation of the activating and interfering effects ensures timely identification of any violations in the humoral regulation of the maternal immune interactions with the HLA semiallogenic fetus, considered as a risk factor for developing sporadic conotruncal heart malformations in the next generation.

1. Bokeriya, L.A., Gudkova R.G., Serdechno-sosudistaya hirurgiya-2014. Bolezni i vrozhdennye anomalii sistemy krovoobrashcheniya. Moscow: NCSSKH im. A.N.Bakuleva; 2015. (In Russian)

2. Eurocat. Available at: httр: //www.eurocat/network.eu. (accessed 10.02.2016).

3. Shabaldin A.V., Glebova L.A., Bachina A.V., Schastlivcev E.L., Potapov V.P. Features of epidemiology of congenital heart diseases at children Kemerovo, as large industrial centre. Complex Issues of Cardiovascular Diseases. 2014;(4):38-46. (In Russian)

4. Garcı́a-Enguı́danos A., Calle M.E., Valero J., Luna S., Domínguez-Rojas V. Risk factors in miscarriage and malformation: a review. European Journal of Obstetrics and Gynecology and Reproductive Biology. 2002; 102(2): 111-119. doi: https://doi.org/10.1016/S0301-2115(01)00613-3

5. Ganu R.S., Harris R.A., Collins K., Aagaard K.M. Early Origins of Adult Disease: Approaches for Investigating the Programmable Epigenome in Humans, Nonhuman Primates, and Rodents ILAR J. 2012; 53 (3-4): 306-321. doi: 10.1093/ilar.53.3-4.306

6. Baack M.L., Wang C., Hu S., Segar J.L., Norris A.W. Hyperglycemia induces embryopathy, even in the absence of systemic maternal diabetes: an in vivo test of the fuel mediated teratogenesis hypothesis. Reprod Toxicol. 2014;46:129–136. doi:10.1016/j.reprotox.2014.03.013.

7. Liddy K.A., White M.Y., Cordwell S.J..Functional decorations: post-translational modifications and heart disease delineated by targeted proteomics Genome Med. 2013; 5(2): 20. DOI: 10.1186/gm424

8. Oyama K., El-Nachef D., Zhang Y., Sdek P. MacLellan W.R. Epigenetic regulation of cardiac myocyte differentiation. Frontiers in Genetics, 2014; 5:375. doi: 10.3389/fgene.2014.00375.

9. Sepiashvili R. I. Funkcional'naya sistema immunnogo gomeostaza Allergologiya i immunologiya. 2015; 16(1): 91-100. (In Russian)

10. Morin-Papunen L., Tiilikainen A., Hartikainen-Sorri A.L. Maternal HLA immunization during pregnancy: presence of anti HLA antibodies in half of multigravidous women. Med Biol. 1984; 62(6): 323-325.

11. Koichi I., Tadao T., Norio T. Possible mechanisms of immunotherapy for maintaining pregnancy in recurrent spontaneous aborters: analysis of anti-idiotypic antibodies directed against autologous T-cell receptors. Human Reproduction. 1999; 14(3): 650–655.

12. Miranda S., Litwin S., Barrientos G. Dendritic cells therapy confers a protective microenvironment in murine pregnancy. Scand J Immunol. 2006; 64(5): 493-499.

13. YArilin A. A. Immunologii: uchebnik. Moscow: GEOTAR-Media; 2010. (In Russian)

14. Inada K., Shima T., Nakashima A. Characterization of regulatory T cells in decidua of miscarriage cases with abnormal or normal fetal chromosomal content. J Reprod Immunol. 2013; 97(1): 104-111. doi: 10.1016/j.jri.2012.12.001;

15. CHistyakova G.N., SHabaldin A.V., Belenkova O.V., Mozes V.G., Matveeva V. G., SHabaldina Е. V., Remizova I. I., Gazieva I. A. Federal'noe gosudarstvennoe byudzhetnoe uchrezhdenie "Ural'skij nauchno-issledovatel'skij institut ohrany materinstva i mladenchestva" Ministerstva zdravoohraneniya Rossijskoj Federacii, patentoobladatel' Sposob opredeleniya allogennogo immunnogo otveta v kratkovremennoj smeshannoj kul'ture limfocitov nerodstvennyh donorov. Patent RF 2581925. 20.02.2016. (In Russian)

16. Sambur M.B. Sposob ocenki vzaimodejstviya limfocitov in vitro, osnovannyj na opredelenii ih rozetkoobrazuyushchej sposobnosti. Immunologiya. 1991; 2: 30-33 (In Russian)

17. Singh M., Orazulike N. C., Ashmore J., Konje J. C. Changes in maternal serum transforming growth factor beta-1 during pregnancy: a cross-sectional study. BioMed research international. 2013;2013:318464 doi: 10.1155/2013/318464.

18. Li Q. Transforming growth factor β signaling in uterine development and function. J Anim Sci Biotechnol. 2014;5(1):52. doi:10.1186/2049-1891-5-52;

19. Chatterjee P., Chiasson V.L., Bounds K.R., Mitchell B.M. Regulation of the Anti-Inflammatory Cytokines Interleukin-4 and Interleukin-10 during Pregnancy. Front Immunol. 2014;5:253. doi:10.3389/fimmu.2014.00253.

20. Shirshev S.V. Gormonal'nye mekhanizmy regulyacii immunnoj sistemy v period beremennosti. Uspekhi sovrem. biologii. 2005; 6: 555-566 (In Russian)

21. Mjosberg J., Berg G., Jenmalm M.C., Ernerudh J. FOXP3+ regulatory T cells and T helper 1, T helper 2, and T helper 17 cells in human early pregnancy deciduas. Biol Reprod. 2010. 82(4): 698-705. doi: 10.1095/biolreprod.109.081208.

22. Haitov R. M., Alekseev L. P., Kofiadi I. A. Rol' immunogenetiki v reshenii fundamental'nyh i prikladnyh zadach personalizirovannoj mediciny. Medicina ekstremal'nyh situacij. 2016; 3:(57): 9-24. (In Russian)

23. Gazieva I.A., Chistyakova G.N., Remizova I.I. Role of cytokine production disorders in genesis of placental insufficiency and early reproductive losses. Medical Immunology (Russia). 2014;16(6):539-550. (In Russian) https://doi.org/10.15789/1563-0625-2014-6-539-550

24. Robertson S.A., Prins J. R., Sharkey D. J., Moldenhauer L.M. Seminal fluid and the generation of regulatory T cells for embryo implantation. Am J Reprod Immunol. 2013; 69(4): 315-303;

25. Belenkova O.V., Mozes V.G., SHabaldin A.V., SHabaldina Е.V. Harakter allogennyh vzaimodejstvij suprugov v kratkovremennoj smeshannoj kul'ture pri immunnyh formah reproduktivnyh poter'. ZHurnal teoreticheskoj i klinicheskoj mediciny (Uzbekistan). 2014; 3(1): 219-222. (In Russian)

26. Belenkova O.V., Mozes V.G., SHabaldin A.V., Lisachenko G.V. Pokazateli allogennyh vzaimodejstvij limfocitov suprugov kak dopolnitel'nye diagnosticheskie i prognosticheskie kriterii immunnyh form reproduktivnyh poter'. Saratovskij nauchno-medicinskij zhurnal. 2013; 4(9): 649-652. (In Russian)