CD39⁺ EXPRESSION BY REGULATORY T CELLS IN PULMONARY SARCOIDOSIS AND LOFGREN’S SYNDROME

Sarcoidosis is a disorder of unknown etiology characterized by development of necrosis-free epithelioid cell granulomas in various tissues. There are two main phenotypes of pulmonary sarcoidosis (PS): Lofgren’s syndrome (LS) is an acute form with favorable outcome, while non-Lofgren’s syndrome (nLS) is a chronic type of disease that can lead to pulmonary fibrosis in 20% of cases.

Our study was aimed at investigating changes in the main cell-surface differentiation antigens on peripheral blood regulatory T cells (Tregs) from the patients with first diagnosed PS without treatment (LS, n = 11) and nLS (n = 46) compared to healthy volunteers (HC, n = 26).

These indexes might be used as immunological markers for predicting severity of this disorder. Flow cytometry analysis of peripheral blood cell samples demonstrated that the nLS patients had decreased relative numbers of CD3⁺ cells vs healthy controls, as well as diminished CD3⁺CD4⁺ cells vs HC and LS patients. Furthermore, the relative and absolute Treg numbers were also decreased in nLS group vs HC (2.83% (2.47; 3.36) vs 3.33% (2.79; 3.84), p = 0.021), and 37 (29; 52) cells vs 50 (42; 65), p = 0.004, respectively) per one microliter of peripheral blood. Relative number of CD39-positive Тregs in chronic vs acute sarcoidosis patients was associated with 51.02% (38.20; 61.62) vs 48.64% (41.46; 63.72) that was significantly (p < 0.001 and p = 0.007, respectively) higher than in HC (39.52% (11.55; 46.34). We have found that “naïve” (CD45R0^-CD62L⁺) Тregs did not significantly differ in percentage of CD39- and CD73-positive cells in all the groups tested. Moreover, CD45R0⁺CD62L⁺ Тregs in LS and nLS patients contained significantly more CD39-positive cells (69.66% (61.92; 79.34) and 67.62% (61.92; 79.34), respectively, compared to 47.55% (15.74; 65.32) in HC (p < 0.001 and p = 0.004, respectively). In case of CD45R0 + CD62LTregs able to exit from the circulation and migrate to the site of inflammation, an increased percentage of CD39-positive subset was noted only in patients with chronic sarcoidosis and HC (61.79% (55.12; 73.09) and 57.27% (16.03; 66.98), p = 0.006). Enhanced CD39 expression on Tregs seems to be related to chronic immune response, so that antigen elimination becomes impossible due to Treg overactivation, as shown in patients with sarcoidosis and some other chronic autoimmune and infectious disorders.

1. Vizel I.Yu. Sarcoidosis: the possibility of spontaneous remission. Vestnik sovremennoy klinicheskoy meditsiny = Bulletin of Contemporary Clinical Medicine, 2012, Vol. 5, no. 2, pp. 54-60. (In Russ.)

2. Vizel I.Yu., Shmelev E.I., Baranova O.P., Barlamov P.N., Borodina G.L., Denisova O.A., Dobin V.L., Kulbaisov A.M., Kupaev V.I., Listopadova M.V., Ovsyannikov N.V., Oskin D.N., Petrov D.V., Solovyov K.I., Shulzhenko L.V., Vizel A.A. Health status of sarcoidosis patients in baseline and in 10 years under different treatment strategies (multicenter analysis). Pulmonologiya = Russian Pulmonology, 2012, Vol. 4, pp. 29-33.(In Russ.)

3. Golovkin A.S., Serebryakova M.K., Zhiduleva E.V., Murtazalieva P.M., Titov V.A., Irtyuga O.B., Moiseeva O.M., Krobinets I.I., Kudryavtsev I.V. Purinergic signaling receptors expression on peripheral T-lymphocytes of healthy donors. Translyatsionnaya meditsina = Translational Medicine, 2017, Vol. 4, no. 5, pp. 46-60.(In Russ.)

4. Golovkin A.S., Asadullina I.A., Kudryavtsev I.V. Purinergic regulation of basic physiological and pathological processes. Meditsinskaya immunologiya = Medical Immunology (Russia), 2018, Vol. 20, no. 4, pp. 463-476. (In Russ.) doi: 10.15789/1563-0625-2018-4-463-476.

5. Zhulai G.A., Oleinik E.K., Churov A.V., Romanov A.A., Kravchenko (Semakova) P.N., Оleinik V.M. Significance of Treg cells for adenosine-mediated immune suppression in colorectal cancer. Meditsinskaya immunologiya = Medical Immunology (Russia), 2017, Vol. 19, no. 1, pp. 89-94.(In Russ.) doi: 10.15789/1563-0625-2017-1-89-94.

6. Ilkovich M.M. Disseminated lung diseases]. Moscow: GEOTAR-Media, 2011. 480 p.

7. Ilkovich M.M., Kokosova A.N. Interstitial lung diseases: a guide for physicians]. St. Petersburg: Nordmedizdat, 2005. 560 p.

8. Kozlov V.A. Suppressor cells – the basis of immunopathogenesis autoimmune diseases. Meditsinskaya immunologiya = Medical Immunology (Russia), 2016, Vol. 18, no. 1, pp. 7-14. (In Russ.) doi:10.15789/1563-0625-2016-1-7-14.

9. Kudryavtsev I.V., Elezov D.S. Analysis of the main populations of cytotoxic T lymphocytes of peripheral blood on the basis level or the expression of CD27, CD28, CD45R0 and CD62L. Rossiyskiy immunologicheskiy zhurnal = Russian Journal of Immunology, 2013, Vol. 7 (16), no. 2-3 (1), pp. 57-61.(In Russ.)

10. Kudryavtsev I.V., Savitsky V.P. Multicolor flow cytometric analysis of the main subpopulations of T-helpers and cytotoxic T cells from peripheral blood. Rossiyskiy immunologicheskiy zhurnal = Russian Journal of Immunology, 2012, Vol. 6, no. 3 (1) (14), pp. 94-99.(In Russ.)

11. Kudryavtsev I.V., Subbotovskaya A.I. Application of six-color flow cytometric analysis for immune profile monitoring. Meditsinskaya immunologiya = Medical Immunology (Russia), 2015, Vol. 17, no. 1, pp. 19-26.(In Russ.) doi: 10.15789/1563-0625-2015-1-19-26.

12. Khaydukov S.V., Baydun L.A., Zurochka A.V., Totolian Areg A. Standardized technology “Research of lymphocytes subpopulation composition in peripheral blood using flow cytometry analyzers” (Draft). Meditsinskaya immunologiya = Medical Immunology (Russia), 2012, Vol. 14, no. 3, pp. 255-268. (In Russ.) doi: 10.15789/1563-0625-2012-3-255-268.

13. Chuchalin A.G., Vizel A.A., Ilkovich M.M., Avdeev S.N., Amirov N.B., Baranova O.P., Borisov S.E., Vizel I.Yu., Lovacheva O.V., Ovsyannikov N.V., Petrov D.V., Romanov V.V., Samsonova M.V., Solovyova I.P., Stepanian I.E., Tyurin I.E., Cherniayev A.L., Shmelev E.I., Shmeleva N.M. Diagnosis and treatment of sarcoidosis: Summary of federal conciliative clinical recommendations (Part II. diagnosis, treatment, prognosis). Vestnik sovremennoy klinicheskoy meditsiny = Bulletin of Contemporary Clinical Medicine, 2014, Vol. 7, no. 5, pp. 73-81.(In Russ.)

14. Broos C.E., van Nimwegen M., Hoogsteden H.C., Hendriks R.W., Kool M., van den Blink B. Granuloma formation in pulmonary sarcoidosis. Front. Immunol., 2013, Vol. 10, no. 4, p. 437.

15. Caramalho I., Nunes-Cabaço H., Foxall R.B., Sousa A.E. Regulatory T-cell development in the human thymus. Front. Immunol., 2015, Vol. 6, p. 395.

16. Facco M., Cabrelle A., Teramo A., Olivieri V., Gnoato M., Teolato S., Ave E., Gattazzo C., Fadini G.P., Calabrese F., Semenzato G., Agostini C. Sarcoidosis is a Th1/Th17 multisystem disorder. Thorax, 2011, Vol. 66, no. 2, pp. 144-150.

17. Fujimura T., Kambayashi Y., Aiba S. Expression of CD39/Entpd1 on granuloma-composing cells and induction of Foxp3-positive regulatory T cells in sarcoidosis. Clin. Exp. Dermatol., 2013, Vol. 38, no. 8, pp. 883-889.

18. Garcia Santana C.A., Tung J.W., Gulnik S. Human Treg cells are characterized by low/negative CD6 expression. Cytometry A, 2014, Vol. 85, no. 10, pp. 901-908.

19. Grant C.R., Liberal R., Mieli-Vergani G., Vergani D., Longhi M.S. Regulatory T-cells in autoimmune diseases: challenges, controversies and yet unanswered questions. Autoimmun. Rev., 2015, Vol. 14, no. 2, pp. 105-116.

20. Huang H., Lu Z., Jiang C., Liu J., Wang Y., Xu Z. Imbalance between Th17 and regulatory T-cells in sarcoidosis. Int. J. Mol. Sci., 2013, Vol. 14, no. 11, pp. 21463-21473.

21. Liu Y., Qiu L., Wang Y., Aimurola H., Zhao Y., Li S., Xu Z. The circulating Treg/Th17 cell ratio is correlated with relapse and treatment response in pulmonary sarcoidosis patients after corticosteroid withdrawal. PLoS ONE, 2016, Vol. 11, no. 2, e0148207. doi: 10.1371/journal.pone.0148207.

22. Loke W.S., Herbert C., Thomas P.S. Sarcoidosis: immunopathogenesis and immunological markers. Int. J. Chronic Dis., 2013, Vol. 2013, 928601. doi: 10.1155/2013/928601.

23. Mahnke Y.D., Roederer M. Optimizing a multicolor immunophenotyping assay. Clin. Lab. Med., 2007, Vol. 27, no. 2, pp. 469-485.

24. Miyara M., Amoura Z., Parizot C., Badoual C., Dorgham K., Trad S., Kambouchner M., Valeyre D., Chapelon-Abric C., Debré P., Piette J.C., Gorochov G. The immune paradox of sarcoidosis and regulatory T cells. J. Exp. Med., 2006, Vol. 203, no. 2, pp. 359-370.

25. Miyara M., Yoshioka Y., Kitoh A., Shima T., Wing K., Niwa A., Parizot C., Taflin C., Heike T., Valeyre D., Mathian A., Nakahata T., Yamaguchi T., Nomura T., Ono M., Amoura Z., Gorochov G., Sakaguchi S. Functional delineation and differentiation dynamics of human CD4 + T-cells expressing the FoxP3 transcription factor. Immunity, 2009, Vol. 30, pp. 899-911.

26. Mortaz E., Rezayat F., Amani D., Kiani A., Garssen J., Adcock I.M., Velayati A. The roles of T helper 1, T helper 17 and regulatory T cells in the pathogenesis of sarcoidosis. Iran J. Allergy Asthma Immunol., 2016, Vol. 15, no. 4, pp. 334-339.

27. Nikolova M., Carriere M., Jenabian M.A., Limou S., Younas M., Kök A., Huë S., Seddiki N., Hulin A., Delaneau O., Schuitemaker H., Herbeck J.T., Mullins J.I., Muhtarova M., Bensussan A., Zagury J.F., Lelievre J.D., Lévy Y. CD39/adenosine pathway is involved in AIDS progression. PLoS Pathog., 2011, Vol. 7, no. 7, e1002110. doi: 10.1371/journal.ppat.1002110.

28. Noack M., Miossec P. Th17 and regulatory T cell balance in autoimmune and inflammatory diseases. Autoimmun. Rev., 2014, Vol. 13, no. 6, pp. 668-677.

29. Nurkhametova D., Kudryavtsev I., Khayrutdinova O., Serebryakova M., Altunbaev R., Malm T., Giniatullin R. Purinergic profiling of regulatory T-cells in patients with episodic migraine. Front. Cell. Neurosci., 2018, Vol. 12, p. 326.

30. Paulissen S.M., van Hamburg J.P., Dankers W., Lubberts E. The role and modulation of CCR6 + Th17 cell populations in rheumatoid arthritis. Cytokine, 2015, Vol. 74, no. 1, pp. 43-53.

31. Rissiek A., Baumann I., Cuapio A., Mautner A., Kolster M., Arck P.C., Dodge-Khatami A., Mittrucker H.W., Koch-Nolte F., Haag F., Tolosa E. The expression of CD39 on regulatory T cells is genetically driven and further upregulated at sites of inflammation. J. Autoimmun., 2015, Vol. 58, pp. 12-20.

32. Saidha S., Sotirchos E.S., Eckstein C. Etiology of sarcoidosis: does infection play a role? Yale J. Biol. Med., 2012, Vol. 85, no. 1, pp. 133-141.

33. Sakaguchi S., Miyara M., Costantino C.M., Hafler D.A. FoxP3 + regulatory T-cells in the human immune system. Nat. Rev. Immunol., 2010, Vol. 10, pp. 490-500.

34. Sakthivel P., Bruder D. Mechanism of granuloma formation in sarcoidosis. Curr. Opin. Hematol., 2017, Vol. 24, no. 1, pp. 59-65.

35. Tang Y., Jiang L., Zheng Y., Ni B., Wu Y. Expression of CD39 on FoxP3 + T regulatory cells correlates with progression of HBV infection. BMC Immunol., 2012, Vol. 13, p. 17.

36. Williams A.E. Immunology mucosal and body surface defences. A.E. Williams. – A. John Willey and Sons, Ltd., Publication, 2015. 380 p.

37. Yegutkin G.G. Enzymes involved in metabolism of extracellular nucleotides and nucleosides: functional implications and measurement of activities. Crit. Rev. Biochem. Mol. Biol., 2014, Vol. 49, no. 6, pp. 473-497.