BACKGROUND: Patients with Sjögren's syndrome, like other patients with autoimmune disorders, display dysregulation in the function of their immune system. Fas and Fas Ligand (FasL) are among the dysregulated proteins. METHODS: We studied Fas and FasL on IL-2Rα+ cells and in serum of patients with Sjögren's syndrome (n = 16) and healthy individuals (n = 16); both from same ethnic and geographical background. We used flow cytometry and enzyme-linked immunosorbent for this purpose. We also measured the expression of Bcl-2 and Bax by reverse transcription quantitative real-time PCR (RT-qPCR) and percentage of apoptotic and dead cells using Annexin V and 7-AAD staining in lymphocytes. RESULTS: FasL was increased in patients' T and B cells while Fas was increased in patients' monocytes, T and B cells. No signs of increased apoptosis were found. sFas and sFasL in patients' serum were increased, although the increase in sFasL was not significant. We suspect an effect of non-steroidal anti-inflammatory therapy on B cells, explaining the decrease of the percentage Fas+ B cells found within our samples. In healthy individuals, there was a noticeable pattern in the expression of FasL which mutually correlated to populations of mononuclear cells; this correlation was absent in the patients with Sjögren's syndrome. CONCLUSIONS: Mononuclear cells expressing IL-2Rα+ had upregulated Fas in Sjögren's syndrome. However, the rate of apoptosis based on Annexin V staining and the Bcl-2/Bax expression was not observed in mononuclear cells. We suspect a functional role of abnormal levels of Fas and FasL which has not been cleared yet.

Department of Biology Jan Evangelista Purkyne University Za Valcovnou 1000 8 400 96 Usti and Labem Czech Republic

Department of Clinical Immunology and Allergology Faculty of Medicine in Hradec Kralove Charles University Simkova 870 500 03 Hradec Kralove Czech Republic

Department of Dentistry Faculty of Medicine in Hradec Kralove Charles University and University Hospital Sokolska 581 500 05 Hradec Kralove Czech Republic

Institute of Clinical Biochemistry and Diagnostics University Hospital Sokolska 581 500 05 Hradec Kralove Czech Republic

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$a Lindrova, Irena $u Department of Dentistry, Faculty of Medicine in Hradec Kralove, Charles University and University Hospital, Sokolska 581, 500 05, Hradec Kralove, Czech Republic $1 https://orcid.org/0000000273273056

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$a Unsolved mystery of Fas: mononuclear cells may have trouble dying in patients with Sjögren's syndrome / $c I. Lindrova, M. Kolackova, T. Svadlakova, R. Vankova, M. Chmelarova, M. Rosecka, E. Jozifkova, M. Sembera, J. Krejsek, R. Slezak

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$a BACKGROUND: Patients with Sjögren's syndrome, like other patients with autoimmune disorders, display dysregulation in the function of their immune system. Fas and Fas Ligand (FasL) are among the dysregulated proteins. METHODS: We studied Fas and FasL on IL-2Rα+ cells and in serum of patients with Sjögren's syndrome (n = 16) and healthy individuals (n = 16); both from same ethnic and geographical background. We used flow cytometry and enzyme-linked immunosorbent for this purpose. We also measured the expression of Bcl-2 and Bax by reverse transcription quantitative real-time PCR (RT-qPCR) and percentage of apoptotic and dead cells using Annexin V and 7-AAD staining in lymphocytes. RESULTS: FasL was increased in patients' T and B cells while Fas was increased in patients' monocytes, T and B cells. No signs of increased apoptosis were found. sFas and sFasL in patients' serum were increased, although the increase in sFasL was not significant. We suspect an effect of non-steroidal anti-inflammatory therapy on B cells, explaining the decrease of the percentage Fas+ B cells found within our samples. In healthy individuals, there was a noticeable pattern in the expression of FasL which mutually correlated to populations of mononuclear cells; this correlation was absent in the patients with Sjögren's syndrome. CONCLUSIONS: Mononuclear cells expressing IL-2Rα+ had upregulated Fas in Sjögren's syndrome. However, the rate of apoptosis based on Annexin V staining and the Bcl-2/Bax expression was not observed in mononuclear cells. We suspect a functional role of abnormal levels of Fas and FasL which has not been cleared yet.

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$a Kolackova, Martina $u Department of Clinical Immunology and Allergology, Faculty of Medicine in Hradec Kralove, Charles University, Simkova 870, 500 03, Hradec Kralove, Czech Republic. kolackovam@lfhk.cuni.cz $1 https://orcid.org/0000000315724565

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$a Svadlakova, Tereza $u Department of Clinical Immunology and Allergology, Faculty of Medicine in Hradec Kralove, Charles University, Simkova 870, 500 03, Hradec Kralove, Czech Republic $1 https://orcid.org/0000000215815531 $7 xx0268729

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$a Vankova, Radka $u Department of Clinical Immunology and Allergology, Faculty of Medicine in Hradec Kralove, Charles University, Simkova 870, 500 03, Hradec Kralove, Czech Republic $1 https://orcid.org/000000028503532X

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$a Chmelarova, Marcela $u Institute of Clinical Biochemistry and Diagnostics, University Hospital, Sokolska 581, 500 05, Hradec Kralove, Czech Republic

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$a Rosecka, Michaela $u Department of Clinical Immunology and Allergology, Faculty of Medicine in Hradec Kralove, Charles University, Simkova 870, 500 03, Hradec Kralove, Czech Republic $1 https://orcid.org/0000000325959990

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$a Jozifkova, Eva $u Department of Biology, Jan Evangelista Purkyne University, Za Valcovnou 1000/8, 400 96, Usti and Labem, Czech Republic $1 https://orcid.org/0000000333543469

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$a Sembera, Martin $u Department of Dentistry, Faculty of Medicine in Hradec Kralove, Charles University and University Hospital, Sokolska 581, 500 05, Hradec Kralove, Czech Republic $1 https://orcid.org/0000000232057090

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$a Krejsek, Jan $u Department of Clinical Immunology and Allergology, Faculty of Medicine in Hradec Kralove, Charles University, Simkova 870, 500 03, Hradec Kralove, Czech Republic $1 https://orcid.org/0000000327786800 $7 nlk19990073429

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$a Slezak, Radovan $u Department of Dentistry, Faculty of Medicine in Hradec Kralove, Charles University and University Hospital, Sokolska 581, 500 05, Hradec Kralove, Czech Republic $1 https://orcid.org/0000000249225062

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