The aim of this study was to determine the effect of autologous serum (AS) eye drops on the density of human leucocyte antigen (HLA)-DR-positive epithelial cells and Langerhans cells on the ocular surface of patients with bilateral severe dry eye disease (DED) due to graft-versus-host disease (GvHD) or Sjögren's syndrome (SS). The study was conducted on 24 patients (48 eyes). AS was applied 6-10 times daily for 3 months together with regular artificial tear therapy. HLA-DR-positive cells were detected by direct immunocytochemistry on upper bulbar conjunctiva imprints obtained before and after treatment. The application of AS drops led to a statistically significant increase in the mean density of aberrant HLA-DR-positive conjunctival epithelial cells (p < 0.05) and HLA-DR-positive Langerhans cells (p < 0.05) in the GvHD group. Aberrant HLA-DR-positive epithelial cells in the SS group were decreased non-significantly. All patients reported a significant decrease in the Ocular Surface Disease Index (p < 0.01), which indicates improvement of the patient's subjective feelings after therapy. There was an expected but non-significant decrease of aberrant HLA-DR-positive conjunctival epithelial cells in the SS group only. However, the increased density of HLA-DR-positive cells, indicating slight subclinical inflammation, does not outweigh the positive effect of AS in patients with DED from GvHD.

Department of Medical Biochemistry Oslo University Hospital Oslo Norway

Department of Ophthalmology 1st Faculty of Medicine Charles University and General University Hospital Prague Prague Czech Republic

Department of Ophthalmology 2nd Faculty of Medicine Charles University Prague and Motol University Hospital and Department of Physiology 2nd Faculty of Medicine Charles University Prague Prague Czech Republic

Department of Plastic and Reconstructive Surgery Oslo University Hospital Oslo Norway

Laboratory of the Biology and Pathology of the Eye Institute of Biology and Medical Genetics 1st Faculty of Medicine Charles University and General University Hospital Prague Prague Czech Republic

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Tsubota K, Goto E, Fujita H, Ono M, Inoue H, Saito I, et al. Treatment of dry eye by autologous serum application in Sjogren's syndrome. British Journal of Ophthalmology. 1999;83(4):390–5. 10.1136/bjo.83.4.390 PubMed DOI PMC

Ogawa Y, Okamoto S, Mori T, Yamada M, Mashima Y, Watanabe R, et al. Autologous serum eye drops for the treatment of severe dry eye in patients with chronic graft-versus-host disease. Bone Marrow Transplant. 2003;31(7):579–83. 10.1038/sj.bmt.1703862 PubMed DOI

Chiang C-C, Lin J-M, Chen W-L, Tsai Y-Y. Allogeneic serum eye drops for the treatment of severe dry eye in patients with chronic graft-versus-host disease. Cornea. 2007;26(7):861–3. 10.1097/ICO.0b013e3180645cd7 PubMed DOI

Na K-S, Kim MS. Allogeneic serum eye drops for the treatment of dry eye patients with chronic graft-versus-host disease. J Ocul Pharmacol Th. 2012;28(5):479–83. PubMed PMC

Azari AA, Karadag R, Kanavi MR, Nehls S, Barney N, Kim K, et al. Safety and efficacy of autologous serum eye drop for treatment of dry eyes in graft-versus-host disease. Cutaneous and Ocular Toxicology. 2017;36(2):152–6. 10.1080/15569527.2016.1209770 PubMed DOI

Geerling G, MacLennan S, Hartwig D. Autologous serum eye drops for ocular surface disorders. British Journal of Ophthalmology. 2004;88(11):1467–74. 10.1136/bjo.2004.044347 PubMed DOI PMC

Bradley JC, Simoni J, Bradley RH, McCartney DL, Brown SM. Time- and temperature-dependent stability of growth factor peptides in human autologous serum eye drops. Cornea. 2009;28(2):200–5. 10.1097/ICO.0b013e318186321e PubMed DOI

Kojima T, Ishida R, Dogru M, Goto E, Matsumoto Y, Kaido M, et al. The effect of autologous serum eyedrops in the treatment of severe dry eye disease: A prospective randomized case-control study. American Journal of Ophthalmology. 2005;139(2):242–6. 10.1016/j.ajo.2004.08.040 PubMed DOI

Goto E, Shimmura S, Shimazaki J, Tsubota K. Treatment of superior limbic keratoconjunctivitis by application of autologous serum. Cornea. 2001;20(8):807–10. 10.1097/00003226-200111000-00006 PubMed DOI

Noble BA, Loh RS, MacLennan S, Pesudovs K, Reynolds A, Bridges L, et al. Comparison of autologous serum eye drops with conventional therapy in a randomised controlled crossover trial for ocular surface disease. British Journal of Ophthalmology. 2004;88(5):647–52. 10.1136/bjo.2003.026211 PubMed DOI PMC

Jirsova K, Brejchova K, Krabcova I, Filipec M, Al Fakih A, Palos M, et al. The application of autologous serum eye drops in severe dry eye patients; subjective and objective parameters before and after treatment. Current Eye Research. 2014;39(1):21–30. 10.3109/02713683.2013.824987 PubMed DOI

Baudouin C, Haouat N, Brignole F, Bayle J, Gastaud P. Immunopathological findings in conjunctival cells using immunofluorescence staining of impression cytology specimens. British Journal of Ophthalmology. 1992;76(9):545–9. 10.1136/bjo.76.9.545 PubMed DOI PMC

Versura P, Profazio V, Schiavi C, Campos EC. Hyperosmolar stress upregulates HLA-DR expression in human conjunctival epithelium in dry eye patients and in vitro models. Invest Ophth Vis Sci. 2011;52(8):5488–96. PubMed

Rolando M, Barabino S, Mingari C, Moretti S, Giuffrida S, Calabria G. Distribution of conjunctival HLA-DR expression and the pathogenesis of damage in early dry eyes. Cornea. 2005;24(8):951–4. 10.1097/01.ico.0000157421.93522.00 PubMed DOI

Baudouin C, Brignole F, Becquet F, Pisella PJ, Goguel A. Flow cytometry in impression cytology specimens—A new method for evaluation of conjunctival inflammation. Invest Ophth Vis Sci. 1997;38(7):1458–64. PubMed

Brignole F, Pisella PJ, Goldschild M, De saint Jean M, Goguel A, Baudouin C. Flow cytometric analysis of inflammatory markers in conjunctival epithelial cells of patients with dry eyes. Invest Ophth Vis Sci. 2000;41(6):1356–63. PubMed

Tsubota K, Fujihara T, Saito K, Takeuchi T. Conjunctival epithelium expression of HLA-DR in dry eye patients. Ophthalmologica Journal international d'ophtalmologie International journal of ophthalmology Zeitschrift für Augenheilkunde. 1999;213(1):16–9. 10.1159/000027387 PubMed DOI

Pisella P-J, Brignole F, Debbasch C, Lozato P-A, Creuzot-Garcher C, Bara J, et al. Flow cytometric analysis of conjunctival epithelium in ocular rosacea and keratoconjunctivitis sicca. Ophthalmology. 2000;107(10):1841–9. 10.1016/s0161-6420(00)00347-x PubMed DOI

Brignole-Baudouin F, Riancho L, Ismail D, Deniaud M, Amrane M, Baudouin C. Correlation between the inflammatory marker HLA-DR and signs and symptoms in moderate to severe dry eye disease. Invest Ophth Vis Sci. 2017;58(4):2438–48. PubMed

Pflugfelder S, Bian F, Gumus K, Farley W, Stern M, De Paiva C. Severity of Sjögren’s syndrome keratoconjunctivitis sicca increases with increased percentage of conjunctival antigen-presenting cells. International Journal of Molecular Sciences. 2018;19(9):2760. PubMed PMC

Epstein SP, Gadaria-Rathod N, Wei Y, Maguire MG, Asbell PA. HLA-DR expression as a biomarker of inflammation for multicenter clinical trials of ocular surface disease. Experimental Eye Research. 2013;111:95–104. 10.1016/j.exer.2013.03.018 PubMed DOI PMC

Jones DT, Monroy D, Ji ZH, Atherton SS, Pflugfelder SC. Sjogrens-syndrome—cytokine and Epstein-Barr viral gene-expression within the conjunctival epithelium. Invest Ophth Vis Sci. 1994;35(9):3493–504. PubMed

Stern ME, Gao JP, Schwalb TA, Ngo ML, Tieu DD, Chan CC, et al. Conjunctival T-cell subpopulations in Sjogren's and non-Sjogren's patients with dry eye. Invest Ophth Vis Sci. 2002;43(8):2609–14. PubMed

Eberwein P, Issleib S, Bohringer D, Mittelviefhaus H, Schwartzkopff J, Finke J, et al. Conjunctival HLA-DR and CD8 expression detected by impression cytology in ocular graft versus host disease. Molecular Vision. 2013;19:1492–501. PubMed PMC

Hanafusa T, Pujolborrell R, Chiovato L, Russell RCG, Doniach D, Bottazzo GF. Aberrant expression of HLA-DR antigen on thyrocytes in Graves-disease—relevance for autoimmunity. Lancet. 1983;2(8359):1111–5. 10.1016/s0140-6736(83)90628-1 PubMed DOI

Ballardini G, Bianchi F, Doniach D, Mirakian R, Pisi E, Bottazzo G. Aberrant expression of HLA-DR antigens on bileduct epithelium in primary biliary cirrhosis: relevance to pathogenesis. Lancet. 1984;324(8410):1009–13. PubMed

Amoils B, Morrison R, Wadee A, Marcus R, Ninin D, King P, et al. Aberrant expression of HLA-DR antigen on valvular fibroblasts from patients with active rheumatic carditis. Clin Exp Immunol. 1986;66(1):88 PubMed PMC

Foulis AK, Farquharson MA. Aberrant expression of HLA-DR antigens by insulin-containing beta-cells in recent-onset type-I Diabetes-Mellitus. Diabetes. 1986;35(11):1215–24. 10.2337/diab.35.11.1215 PubMed DOI

Feng J, Li M, Gu W, Tang H, Yu S. The aberrant expression of HLA-DR in intrahepatic bile ducts in patients with biliary atresia: an immunohistochemistry and immune electron microscopy study. Journal of Pediatric Surgery. 2004;39(11):1658–62. 10.1016/j.jpedsurg.2004.07.010 PubMed DOI

Oberhuber G, Püspök A, Peck-Radosavlevic M, Kutilek M, Lamprecht A, Chott A, et al. Aberrant esophageal HLA-DR expression in a high percentage of patients with Crohn's disease. The American journal of surgical pathology. 1999;23(8):970 10.1097/00000478-199908000-00016 PubMed DOI

Bottazzo GF, Pujolborrell R, Hanafusa T, Feldmann M. Role of aberrant HLA-DR expression and antigen presentation in induction of endocrine autoimmunity. Lancet. 1983;2(8359):1115–9. 10.1016/s0140-6736(83)90629-3 PubMed DOI

Zschaler U, Weber P, Jedrychowski A, Wiedmann KH. Immunohistological analysis of differential HLA expression in patients with various bowel diseases. Zeitschrift Fur Gastroenterologie. 1991;29(3):92–100. PubMed

Appleton A, Sviland L, Pearson A, Wilkes J, Green M, Malcolm A. Diagnostic features of transfusion associated graft versus host disease. J Clin Pathol. 1994;47(6):541–6. 10.1136/jcp.47.6.541 PubMed DOI PMC

Lange A, Klimczak A, Karabon L, Suchnicki K. Cytokines, adhesion molecules (E-selectin and VCAM-1) and graft-versus-host disease. Arch Immunol Ther Exp (Warsz). 1995;43(2):99. PubMed

Tersigni C, Redman C, Dragovic R, Tannetta D, Scambia G, Di Simone N, et al. HLA-DR is aberrantly expressed at feto-maternal interface in pre-eclampsia. J Reprod Immunol. 2018. PubMed

Pflugfelder SC, Geerling G, Kinoshita S, Lemp MA, McCulley J, Nelson D, et al. Management and therapy of dry eye disease: Report of the Management and Therapy Subcommittee of the international Dry Eye WorkShop (2007). Ocular Surface. 2007;5(2):163–78. 10.1016/s1542-0124(12)70085-x PubMed DOI

Schiffman RM, Christianson MD, Jacobsen G, Hirsch JD, Reis BL. Reliability and validity of the ocular surface disease index. Archives of Ophthalmology. 2000;118(5):615–21. 10.1001/archopht.118.5.615 PubMed DOI

Donisi PM, Rama P, Fasolo A, Ponzin D. Analysis of limbal stem cell deficiency by corneal impression cytology. Cornea. 2003;22(6):533–8. 10.1097/00003226-200308000-00009 PubMed DOI

Jirsova K, Juklova K, Vesela V, Filipec M. Morphological and immunocytochemical characterization of snake-like chromatin cells. Histology and Histopathology. 2006;21(4/6):355. PubMed

Pan Q, Angelina A, Marrone M, Stark WJ, Akpek EK. Autologous serum eye drops for dry eye. Cochrane Db Syst Rev. 2017(2). PubMed PMC

Ali TK, Gibbons A, Cartes C, Zarei-Ghanavati S, Gomaa M, Gonzalez I, et al. Use of autologous serum tears for the treatment of ocular surface disease from patients with systemic autoimmune diseases. American journal of ophthalmology. 2018;189:65–70. 10.1016/j.ajo.2018.02.009 PubMed DOI

Rocha E, Pelegrino F, De Paiva C, Vigorito A, De Souza C. GVHD dry eyes treated with autologous serum tears. Bone Marrow Transplant. 2000;25(10):1101 10.1038/sj.bmt.1702334 PubMed DOI

Tahmaz V, Gehlsen U, Sauerbier L, Holtick U, Engel L, Radojska S, et al. Treatment of severe chronic ocular graft-versus-host disease using 100% autologous serum eye drops from a sealed manufacturing system: a retrospective cohort study. British Journal of Ophthalmology. 2017;101(3):322–6. 10.1136/bjophthalmol-2015-307666 PubMed DOI

Nicolle P, Liang H, Reboussin E, Rabut G, Warcoin E, Brignole-Baudouin F, et al. Proinflammatory markers, chemokines, and enkephalin in patients suffering from dry eye disease. International Journal of Molecular Sciences. 2018;19(4):1221. PubMed PMC

Pflugfelder SC, Jones D, Ji ZH, Afonso A, Monroy D. Altered cytokine balance in the tear fluid and conjunctiva of patients with Sjogren's syndrome keratoconjunctivitis sicca. Current Eye Research. 1999;19(3):201–11. 10.1076/ceyr.19.3.201.5309 PubMed DOI

Lee SY, Han SJ, Nam SM, Yoon SC, Ahn JM, Kim TI, et al. Analysis of tear cytokines and clinical correlations in Sjogren syndrome dry eye patients and non-Sjogren syndrome dry eye patients. American Journal of Ophthalmology. 2013;156(2):247–53. 10.1016/j.ajo.2013.04.003 PubMed DOI

Westekemper H, Meller S, Citak S, Schulte C, Steuhl KP, Homey B, et al. Differential chemokine expression in chronic GVHD of the conjunctiva. Bone Marrow Transplant. 2010;45(8):1340–6. 10.1038/bmt.2009.346 PubMed DOI

Brignole F, De Saint-Jean M, Goldschild M, Becquet F, Goguel A, Baudouin C. Expression of Fas-Fas ligand antigens and apoptotic marker APO2 center dot 7 by the human conjunctival epithelium. Positive correlation with class II HLA DR expression in inflammatory ocular surface disorders. Experimental Eye Research. 1998;67(6):687–97. 10.1006/exer.1998.0566 PubMed DOI

Murphy GF, Korngold R. Significance of selectively targeted apoptotic rete cells in graft-versus-host disease. Biol Blood Marrow Tr. 2004;10(6):357–65. PubMed

Imamura M, Hashino S, Kobayashi H, Kubayashi S, Hirano S, Minagawa T, et al. Serum cytokine levels in bone-marrow transplantation—synergistic interaction of Interleukin-6, Interferon-Gamma, and Tumor-Necrosis-Factor-alpha in graft-versus-host disease. Bone Marrow Transplant. 1994;13(6):745–51. PubMed

Szodoray P, Alex P, Jonsson MV, Knowlton N, Dozmorov I, Nakken B, et al. Distinct profiles of Sjogren's syndrome patients with ectopic salivary gland germinal centers revealed by serum cytokines and BAFF. Clin Immunol. 2005;117(2):168–76. 10.1016/j.clim.2005.06.016 PubMed DOI

Oh JY, Kim MK, Choi HJ, Ko JH, Kang EJ, Lee HJ, et al. Investigating the relationship between serum interleukin-17 levels and systemic immune-mediated disease in patients with dry eye syndrome. Korean journal of ophthalmology. 2011;25(2):73–6. 10.3341/kjo.2011.25.2.73 PubMed DOI PMC