Peripheral blood B cell subsets from patients with various activity of chronic sarcoidosis

Sarcoidosis is an inflammatory disease of unknown etiology, characterized by development of necrosis-free epithelioid cell granulomas, resulting in hyperactivation of various cells of the immune system. The role of humoral mechanisms in the pathogenesis of sarcoidosis is less studied than cell-mediated. It is necessary to study the role of activation or the anergy of the B cell development of immunity in sarcoidosis, the degree of its activity and the characteristics of the clinical course of the disease. Our study was aimed at investigating the characteristics of the B cells subsets in the peripheral blood of patients with chronic sarcoidosis (n = 41), depending on the activity of the disease. The control was peripheral blood samples from healthy volunteers (n = 43). Objective clinical and instrumental criteria, angiotensin converting enzyme (ACE) were used to determine the activity of the disease. Using flow cytometry analysis of peripheral blood cell B cells were determined based on two approaches: expression of IgD/CD38 (“Bm1-Bm5” classification) and IgD/CD27. In patients with sarcoidosis there was a significantly higher relative number of Bm2 "activated" naive cells" (IgD⁺CD38⁺) than in conditionally healthy volunteers, 65.38% versus 55,66% (p < 0.001). The relative and absolute contents of eBm5 (IgD^-CD38⁺) and Bm5 (IgD^-CD38⁺) memory cells were significantly lower in the group of patients with sarcoidosis relative to the control group. Relative values: 6.59% versus 13.31%, (p < 0.001), and 3.43% versus 8.49%, (p < 0.001), respectively. It was shown that with an increased level of ACE in the peripheral blood of patients, the number of naive Bm1 cells (IgD⁺CD38^-) was significantly reduced, r = -0.557, p < 0.001. The relative content of memory B cells that did not switch the class of synthesized antibodies (IgD⁺CD27⁺) in the group of patients was reduced to 6,25%, and in the control group — 12,95% (p<0.001). The number of memory cells that switched the class of synthesized antibodies (IgD^-CD27⁺) was also significantly reduced in patients with sarcoidosis and amounted to 6.75% versus 16.50% in the control group (p < 0.001). In patients with high levels of ACE, there was an increase in the relative content of naive B cells (IgD⁺CD27^-), r = 0.532, p < 0.001. An inverse relationship was established between the number of memory B cells (IgD⁺CD27⁺) and ACE levels, r = -0.565, p < 0.001. These results indicate the important role of the B cell immune response in the pathogenesis of sarcoidosis and make it possible to evaluate the characteristics of the humoral response with various degrees of disease activity.

1. Aisanov Z.R., Amirov N.B., Baranova O.P., Vizel А.А., Vizel I.Yu., Gurileva M.E., Fatkullin I.F., Chuchalin A.G. Sarcoidosis. Moscow: “Atmosphera”, 2010, 416 p. (In Russ.). https://elibrary.ru/item.asp?id=36987922

2. Akopov A.L., Baranova O.P., Bogdanov A.N., Bondarenko I.B., Dvorakovskaya I.V., Ilkovich M.M., Ilkovich Yu.M., Luchkevich V.S., Novikova L.N., Orlova G.P., Orlov S.V. Interstitial and orphan lung diseases. Moskow: GEOTAR-Media, 2016, 560 p. (In Russ.). https://elibrary.ru/item.asp?id=26227541

3. Budkova A.I., Lapin S.V., Serebriakova M.K., Kudryavtsev I.V., Trishina I.N., Maslyansky A.L., Totolian A.A. B-cell subpopulations of peripheral blood in systemic lupus erythematosus. Medical Immunology (Russia), 2017, Vol. 19, no. 2, pp. 175-184. (In Russ.). https://doi.org/10.15789/1563-0625-2017-2-175-184.

4. 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). Medical Immunology (Russia), 2012, Vol. 14, no. 3, pp. 255-268. (In Russ.). http://dx.doi.org/10.15789/1563-0625-2012-3-255-268.

5. Ando M., Goto A., Takeno Y., Yamasue M., Komiya K., Umeki K., Nureki S.I., Miyazaki E., Kadota J.I. Significant elevation of the levels of B-cell activating factor (BAFF) in patients with sarcoidosis. Clin Rheumatol., 2018, Vol. 37, no. 10, pp. 2833-2838. doi: 10.1007/s10067-018-4183-2.

6. Beccastrini E., Vannozzi L., Bacherini D., Squatrito D., Emmi L. Successful treatment of ocular sarcoidosis with rituximab. Ocul Immunol Inflamm., 2013, Vol. 21, no. 3, pp. 244-246. DOI: 10.3109/09273948.2012.762982.

7. Beijer E., Veltkamp M., Meek B., Moller D.R.. Etiology and Immunopathogenesis of Sarcoidosis: Novel Insights. Semin Respir Crit Care Med., 2017, Vol. 38, no. 4, pp. 404-416. doi: 10.1055/s-0037-1603087.

8. Belhomme N., Jouneau S., Bouzillé G., Decaux O., Lederlin M., Guillot S., Perlat A., Jégo P. Role of serum immunoglobulins for predicting sarcoidosis outcome: A cohort study. PLoS One, 2018, Vol. 13, no. 4, pp. e0193122. DOI: 10.1371/journal.pone.0193122.

9. Belkhou A., Younsi R., Bouchti I., Hassani S. Rituximab as a treatment alternative in sarcoidosis. Joint Bone Spine, 2008, Vol. 75, pp. 511–512. DOI: 10.1016/j.jbspin.2008.01.025.

10. Bernstein K.E., Ong F.S., Blackwell W.L., Shah K.H., Giani J.F., Gonzalez-Villalobos R.A., Shen X.Z., Fuchs S., Touyz R.M. A modern understanding of the traditional and nontraditional biological functions of angiotensin-converting enzyme. Pharmacol Rev., 2012, Vol. 65, pp. 1-46. doi: 10.1124/pr.112.006809.

11. Bomprezzi R., Pati S., Chansakul C., Vollmer T. A case of neurosarcoidosis successfully treated with rituximab. Neurology, 2010, Vol. 75, pp. 568–570. doi: 10.1212/WNL.0b013e3181ec7ff9.

12. 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, pp. 437. doi: 10.3389/fimmu.2013.00437.

13. Cannon R.O., Cunnion R.E., Parrillo J.E., Palmeri S.T., Tucker E.E., Schenke W.H., Epstein S.E. Dynamic limitation of coronary vasodilator reserve in patients with dilated cardiomyopathy and chest pain. J. Am. Coll. Cardiol., 1987, Vol. 10, pp. 190-200. doi: 10.1016/s0735-1097(87)80118-3.

14. Casanova N., Zhou T., Knox K.S., Garcia J.G. Identifying novel biomarkers in sarcoidosis using genome-based approaches. Clin Chest Med., 2015, Vol. 36, pp. 621-630. doi: 10.1016/j.ccm.2015.08.005.

15. Chopra A., Kalkanis A., Judson M.A. Biomarkers in sarcoidosis. Expert Rev Clin Immunol., 2016, Vol. 12, pp. 1191-1208. doi: 10.1080/1744666X.2016.1196135.

16. Cinetto F., Compagno N., Scarpa R., Malipiero G., Agostini C. Rituximab in refractory sarcoidosis: a single centre experience. Clin Mol Allergy, 2015, Vol. 13, no. 1, pp. 19. doi: 10.1186/s12948-015-0025-9.

17. Csongrádi A., Enyedi A., Takács I., Végh T., Mányiné I.S., Pólik Z., Altorjay I.T., Balla J., Balla G., Édes I., Kappelmayer J., Tóth A., Papp Z., Fagyas M. Optimized angiotensin-converting enzyme activity assay for the accurate diagnosis of sarcoidosis. Clin Chem Lab Med., 2018, Vol. 56, no. 7, pp. 1117-1125. doi: 10.1515/cclm-2017-0837.

18. Dasilva V., Breuil V., Chevallier P., Euller-Ziegler L. Relapse of severe sarcoidosis with an uncommon peritoneal location after TNFalpha blockade. Efficacy of rituximab, report of a single case. Joint Bone Spine, 2010, Vol. 77, pp. 82–83. doi: 10.1016/j.jbspin.2009.11.005.

19. Duan J., Xu Y., Zhu H., Zhang H., Sun S., Sun H., Wang W., Xie S. Relationship between CT activity score with lung function and the serum angiotensin converting enzyme in pulmonary sarcoidosis on chest HRCT. Medicine (Baltimore), 2018, Vol. 97, no. 36, pp e12205. doi: 10.1097/MD.0000000000012205.

20. Georas S.N., Chapman T.J., Crouser E.D. Sarcoidosis and T-helper cells. Th1, Th17, or Th17.1? Am. J. Respir. Crit. Care Med., 2016, Vol. 193, no. 11, pp. 1198–200. doi: 10.1164/rccm.201512-2419ED.

21. Grosso S., Margollicci M.A., Bargagli E., Buccoliero Q.R., Perrone A., Galimberti D., Morgese G., Balestri P., Rottoli P. Serum levels of chitotriosidase as a marker of disease activity and clinical stage in sarcoidosis. Scand J Clin Lab Invest., 2004, Vol. 64, pp. 57-62. doi: 10.1080/00365510410004092.

22. Grutters J.C., Fellrath J.-M., Mulder L., Janssen R., van den Bosch J.M., van Velzen-Blad H. Serum soluble interleukin-2 receptor measurement in patients with sarcoidosis: a clinical evaluation. Chest, 2003, Vol. 124, no. 1, pp. 186-195. doi: 10.1378/chest.124.1.186.

23. Häggmark A., Hamsten C., Wiklundh E., Lindskog C., Mattsson C., Andersson E., Lundberg I.E., Grönlund H., Schwenk J.M., Eklund A., Grunewald J., Nilsson P. Proteomic profiling reveals autoimmune targets in sarcoidosis. Am J Respir Crit Care Med., 2015, Vol. 191, no. 5, pp. 574-583. doi: 10.1164/rccm.201407-1341OC.

24. 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. doi: 10.3390/ijms141121463.

25. Hunninghake G.W., Crystal R.G. Mechanisms of hypergammaglobulinemia in pulmonary sarcoidosis. Site of increased antibody production and role of T lymphocytes. J Clin Invest., 1981, Vol. 67, no. 1, pp. 86-92. doi: 10.1172/JCI110036.

26. Janiak P., Libert O., Vilaine J. P. Role of the renin-angiotensin system in neointima formation after injury in rabbits. Hypertension, 1994, Vol. 24, pp. 671-678. DOI: 10.1161/01.hyp.24.6.671.

27. Kahkouee S., Samadi K., Alai A., Abedini A., Rezaiian L. Serum ACE Level in Sarcoidosis Patients with Typical and Atypical HRCT Manifestation. Pol J Radiol., 2016, Vol. 81, pp. 458-461. doi: 10.12659/PJR.897708.

28. Kobak S., Yilmaz H., Sever F., Duran A., Sen N., Karaarslan A. The prevalence of antinuclear antibodies in patients with sarcoidosis. Autoimmune Dis., 2014, Vol. 2014, pp. 351852. doi: 10.1155/2014/351852.

29. Kobak S., Ylmaz H., Sever F., Duran A., Sen N. Anti-cyclic citrullinated peptide antibodies in patients with sarcoidosis. Sarcoidosis Vasc Diffuse Lung Dis., 2014, Vol. 31, no. 3, pp. 206-210. https://www.ncbi.nlm.nih.gov/pubmed/25363220.

30. Krause M.L., Cooper L.T., Chareonthaitawee P., Amin S. Successful use of rituximab in refractory cardiac sarcoidosis. Rheumatology (Oxford), 2016, Vol. 55, no. 1, pp. 189–191. doi: 10.1093/rheumatology/kev309.

31. Lee N.S., Barber L., Akula S.M., Sigounas G., Kataria Y.P., Arce S. Disturbed homeostasis and multiple signaling defects in the peripheral blood B-cell compartment of patients with severe chronic sarcoidosis. Clin Vaccine Immunol., 2011, Vol. 18, no. 8, pp. 1306-1316. doi: 10.1128/CVI.05118-11.

32. 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, pp. e0148207. doi: 10.1371/journal.pone.0148207.

33. Mathur R.S., Kurdikar V.L., Shah J.R. Serum angiotensin converting enzyme in the diagnosis of pulmonary sarcoidosis. J Assoc Physicians India, 1990, Vol. 38, pp. 474–475. https://www.ncbi.nlm.nih.gov/pubmed/1963426.

34. Miedema J.R., Kaiser Y., Broos C.E., Wijsenbeek M.S., Grunewald J., Kool M. Th17-lineage cells in pulmonary sarcoidosis and Löfgren's syndrome: Friend or foe? J Autoimmun., 2018, Vol. 87, pp. 82-96. doi: 10.1016/j.jaut.2017.12.012.

35. O'Reilly P.J., Ding Q., Akthar S., Cai G., Genschmer K.R., Patel D.F., Jackson P.L., Viera L., Roda M., Locy M.L., Bernstein E.A., Lloyd C.M., Bernstein K.E., Snelgrove R.J., Blalock J.E. Angiotensin-converting enzyme defines matrikine-regulated inflammation and fibrosis. JCI Insight, 2017, Vol. 2, nn. 22, pp. 91923. doi: 10.1172/jci.insight.91923.

36. Ors F., Gumus S., Aydogan M., Sari S., Verim S., Deniz O. HRCT findings of pulmonary sarcoidosis; relation to pulmonary function tests. Multidiscip Respir Med., 2013, Vol. 8, pp. 8. doi: 10.1186/2049-6958-8-8.

37. Palchevskiy V., Hashemi N., Weigt S.S., Xue Y.Y., Derhovanessian A., Keane M.P., Strieter R.M., Fishbein M.C., Deng J.C., Lynch J.P., Elashoff R., Belperio J.A. Immune response CC chemokines CCL2 and CCL5 are associated with pulmonary sarcoidosis. Fibrogenesis Tissue Repair, 2011, Vol. 4, pp. 10. doi: 10.1186/1755-1536-4-10.

38. Popević S., Šumarac Z., Jovanović D., Babić D., Stjepanović M., Jovičić S., Šobić-Šaranović D., Filipović S., Gvozdenović B., Omčikus M., Milovanović A., Videnović-Ivanov J,, Radović A,, Žugić V., Mihailović-Vučinić V. Verifying sarcoidosis activity: chitotriosidase versus ace in sarcoidosis— case-control study. J Med Biochem, 2016, Vol. 35, pp. 390-400. doi: 10.1515/jomb-2016-0017.

39. Rauch M., Tussiwand R., Bosco N., Rolink A.G. Crucial role for BAFF-BAFF-R signaling in the survival and maintenance of mature B cells. PLoS One, 2009, Vol. 4, pp. e5456. doi: 10.1371/journal.pone.0005456.

40. Sakthivel P., Bruder D. Mechanism of granuloma formation in sarcoidosis. Curr. Opin. Hematol., 2017, Vol. 24, no. 1, pp. 59-65. doi: 10.1097/MOH.0000000000000301.

41. Saussine A., Tazi A., Feuillet S., Rybojad M., Juillard C., Bergeron A., Dessirier V., Bouhidel F., Janin A., Bensussan A., Bagot M., Bouaziz J.D. Active chronic sarcoidosis is characterized by increased transitional blood B cells, increased IL-10-producing regulatory B cells and high BAFF levels. PLoS One, 2012, Vol. 7, no. 8, pp. 43588. doi: 10.1371/journal.pone.0043588.

42. Stohl W., Xu D., Kim K.S., Koss M.N., Jorgensen T.N., Deocharan B., Metzger T.E., Bixler S.A., Hong Y.S., Ambrose C.M., Mackay F., Morel L., Putterman C., Kotzin B.L., Kalled S.L. BAFF overexpression and accelerated glomerular disease in mice with an incomplete genetic predisposition to systemic lupus erythematosus. Arthritis Rheum., 2005, Vol. 52, pp. 2080–2091. doi: 10.1002/art.21138.

43. Ten Berge B., Paats M.S., Bergen I.M., van den Blink B., Hoogsteden H.C., Lambrecht B.N., Hendriks R.W., Kleinjan A. Increased IL-17A expression in granulomas and in circulating memory T cells in sarcoidosis. Rheumatology (Oxford), 2012, Vol. 51, no. 1, pp. 37-46. doi: 10.1093/rheumatology/ker316.

44. Ueda-Hayakawa I., Tanimura H., Osawa M., Iwasaka H., Ohe S., Yamazaki F., Mizuno K., Okamoto H. Elevated serum BAFF levels in patients with sarcoidosis: association with disease activity. Rheumatology (Oxford), 2013, Vol. 52, no. 9, pp. 1658–1666. doi: 10.1093/rheumatology/ket186.

45. Vorselaars A.D., van Moorsel C.H., Zanen P., Ruven H.J., Claessen A.M., van Velzen-Blad H., Grutters J.C. ACE and sIL-2R correlate with lung function improvement in sarcoidosis during methotrexate therapy. Respir Med., 2015, Vol. 109, pp. 279-285. doi: 10.1016/j.rmed.2014.11.009.

46. Weinberg I., Vasiliev L., Gotsman I. Anti-dsDNA antibodies in sarcoidosis. Semin Arthritis Rheum., 2000, Vol. 29, no. 5, pp. 328-331. https://www.ncbi.nlm.nih.gov/pubmed/10805357.

47. Weir E.K. Hypoxic pulmonary vasoconstriction. Chest, 1982, Vol. 82., pp. 519-521.