Research of subpopulation CD3+ CD4- CD8- double-negative T lymphocytes in kidney transplant recipients

Introduction. CD3⁺CD4^- CD8^- cells represent one of the subpopulations of T-regulatory lymphocytes. According to literature reports, an increase in the content of graft-infiltrating CD3⁺CD4^- CD8^- T cells was detected in the heart xenograft tissues of an experimental model with a long-term graft survival. The efficacy of CD3⁺CD4^- CD8^- infusion to induce skin graft tolerance was described. Some studies have shown that a decrease in CD3⁺CD4^- CD8^- content in peripheral blood of patients undergoing to hematopoietic stem cell transplantation was associated with the development of a graft-versus-host reaction.

Objectives. To study changes in the values of CD3⁺CD4^- CD8^- double-negative T lymphocytes in peripheral blood in kidney transplant recipients.

Material and methods. The study included 165 recipients who underwent kidney transplantation. The creatinine and urea concentrations in blood were determined before surgery, on day 7, and day 360 after transplantation. The content of CD3⁺CD4^- CD8^- lymphocytes was studied before surgery, on the 3rd, 7th, 30th, 90th, 180th and 360th days after surgery. Early graft function was assessed on day 7 after transplantation. The function was defined as a primary one at creatinine levels below 300 μmol/L. The graft dysfunction was defined as creatinine values equal to or greater than 300 μmol/L and the need for dialysis in the first week after surgery. The satisfactory graft function after a year was characterized by the blood creatinine level below 150 μmol/L, absent episodes of graft rejection, and no need for dialysis in the first year of follow-up. There were 4 groups of recipients formed. The first group included patients with the primary graft function and satisfactory late graft function. The second group included patients with the primary function and late graft dysfunction. The third group included patients with the primary dysfunction and late satisfactory function. The fourth group included patients with the primary and late graft dysfunction.

Results. In the first and second groups, there were no significant differences in the blood level of CD3⁺CD4^- CD8^- during the year. After a year, a significant CD3⁺CD4^- CD8^- decrease was noted in the group with late graft dysfunction. A similar tendency was revealed in the third and fourth groups. In the fourth group (with late graft dysfunction), the level of CD3⁺CD4^- CD8^- was significantly lower only after a year of observation compared with the levels in the third group. A negative correlation was noted between the CD3⁺CD4^- CD8^- values and the creatinine and urea levels. Thus, high CD3⁺CD4^- CD8^- values in kidney transplant recipients after a year were associated with a satisfactory graft function.

Conclusions. 1. A stable 1-year satisfactory kidney graft function is characterized by an increase in the blood level of CD3⁺CD4^- CD8^- T lymphocytes. 2. A kidney graft dysfunction in the late post-transplant period is characterized by a decrease in the blood level of CD3⁺CD4^- CD8^- T lymphocytes.

Authors declare no conflict of interest.

1. Gao JF, McIntyre MS, Juvet SC, Diao J, Li X, Vanama RB, et al. Regulation of antigen-expressing dendritic cells by double negative regulatory T cells. Eur J Immunol. 2011;41(9):2699– 708. PMID: 21660936 https://doi.org/10.1002/eji.201141428

2. Sakaguchi S, Miyara M, Costantino CM, Hafler DA. FOXP3+ regulatory T cells in the human immune system. Nat Rev Immunol. 2010;10(7):490–500. PMID: 20559327 https://doi.org/10.1038/nri2785

3. Zhang Z, Yang L, Young K, DuTempl e B, Zhang L. Identification of a previously unknown antigen-specific regulatory T cell and its mechanism of suppression. Nat Med. 2000;6(7):782–789. https://doi.org/10.1038/77513

4. Brandt D, Hedrich CM. TCR(+) CD3(+)CD4(-)CD8(-) (double negative) T cells in autoimmunity. Autoimmun Rev. 2018;17(4):422–430. PMID: 29428806 https://doi.org/10.1016/j. autrev.2018.02.001

5. Ford MS, Chen W, Wong S, Li C, Vanama R, Elford AR, et al. Peptide-activated double-negative T cells can prevent autoimmune type-1 diabetes development. Eur J Immunol. 2007;37(8):2234– 41. PMID: 17578845 https://doi.org/10.1002/eji.200636991

6. Chen W, Ford MS, Young KJ, Cybulsky MI, Zhang L. Role of double-negative regulatory T cells in long-term cardiac xenograft survival. J Immunol. 2003;170(4):1846–53. PMID: 12574350 https://doi.org/10.4049/jimmunol.170.4.1846

7. Voelkl S, Gary R, Mackensen A. Characterization of the immunoregulatory function of human TCR-+ CD4- CD8- double-negative T cells. Eur J Immunol. 2011;41(3):739–48. PMID: 21287552 https://doi.org/10.1002/eji.201040982

8. Zhang D, Yang W, Degauque N, Tian Y, Mikita A, Zheng XX. New differentiation pathway for double-negative regulatory T cells that regulates the magnitude of immune responses. Blood. 2007;109(9):4071–9. PMID: 17197428 https://doi.org/10.1182/blood-2006-10- 050625

9. Chen W, Zhou D, Torrealba JR, Waddell TK, Grant D, Zhang L. Donor lymphocyte infusion induces long-term donor-specific cardiac xenograft survival through activation of recipient double-negative regulatory T cells. J Immunol. 2005;175(5):3409–16. PMID: 16116235 https://doi.org/10.4049/jimmunol.175.5.3409

10. Hillhouse EE, Beauchamp C, ChabotRoy G, Dugas V, Lesage S. Interleukin-10 limits the expansion of immunoregulatory CD4-CD8- T cells in autoimmuneprone non-obese diabetic mice. Immunol Cell Biol. 2010;88(8):771–80. PMID: 20603635 https://doi.org/10.1038/icb.2010.84

11. Zhang ZX, Ma Y, Wang H, Arp J, Jiang J, Huang X, et al. Double-negative T cells, activated by xenoantigen, lyse autologous B and T cells using a perforin/granzyme-dependent, FasFas ligand-independent pathway. J Immunol. 2006;177(10):6920–9. PMID: 17082607 https://doi.org/10.4049/jimmunol.177.10.6920

12. Chen W, Diao J, Stepkowski SM, Zhang L. Both infiltrating regulatory T cells and insufficient antigen presentation are involved in long-term cardiac xenograft survival. J Immunol. 2007;179(3):1542–8. PMID: 17641020 https://doi.org/10.4049/jimmunol.179.3.1542

13. Chen W, Ford MS, Young KJ, Zhang L. Infusion of in vitro-generated DN T regulatory cells induces permanent cardiac allograft survival in mice. Transplant Proc. 2003;35(7):2479–80. PMID: 14611991 https://doi.org/10.1016/j. transproceed.2003.08.030

14. Young KJ, DuTemple B, Phillips MJ, Zhang L. Inhibition of graft-versus-host disease by double-negative regulatory T cells. J Immunol. 2003;171(1):134– 41. PMID: 12816991 https://doi.org/10.4049/jimmunol.171.1.134

15. McIver Z, Serio B, Dunbar A, O'Keefe CL, Powers J, Wlodarski M, et al. Double-negative regulatory T cells induce allotolerance when expanded after allogeneic haematopoietic stem cell transplantation. Br J Haematol. 2008;141(2):170–8. PMID: 18318770 https://doi.org/10.1111/j.1365-2141

16. Young KJ, Kay LS, Phillips MJ, Zhang L. Antitumor activity mediated by double-negative T cells. Cancer Res. 2003;63(22):8014–21. PMID: 14633734.

17. Sakaguchi S, Yamaguchi T, Nomura T, Ono M. Regulatory T cells and immune tolerance. Cell. 2008;133(5):775– 787. PMID: 18510923 https://doi.org/10.1016/j.cell.2008.05.009

18. Chen W, Ford MS, Young KJ, Zhang L. The role and mechanisms of double negative regulatory T cells in the suppression of immune responses. Cell Mol Immunol. 2004;1(5):328–35. PMID: 16285891.

19. Ford McIntyre MS1, Gao JF, Li X, Naeini BM, Zhang L. Consequences of double negative regulatory T cell and antigen presenting cell interaction on immune response suppression. Int Immunopharmacol. 2011;11(5):597– 603. https://doi.org/10.1016/j. intimp.2010.11.015

20. Cantaluppi V, Dellepiane S, Tamagnone M, Medica D, Figliolini F, Messina M, et al. Neutrophil gelatinase associated lipocalin is an early and accurate biomarker of graft function and tissue regeneration in kidney transplantation from extended criteria donors. PLoS One. 2015;10(6):e0129279. PMID: 26125566 https://doi.org/10.1371/journal.pone.0129279

21. Massart A, Ghisdal L, Abramo wicz M, Abramowicz D. Operational tolerance in kidney transplantation and associated biomarkers. Clin Exp Immunol. 2017;189(2):138–157. PMID: 28449211 https://doi.org/10.1111/cei.12981