SAT0486 TNF Inhibitors Have No Effects To Human T-Lymphotropic Virus Type-I (HTLV-I)-Infected Cell Line. (15th July 2016)

• Record Type: Journal Article
• Authors: Fukui, S.; Nakamura, H.; Takahashi, Y.; Hasegawa, H.; Yanagihara, K.; Nakamura, T.; Kawakami, A.
• Is Part Of: Annals of the rheumatic diseases. Volume 75(2016)Supplement 2
• Page Start: 846
• Page End: 847
• Publication Date: 2016-07-15
• Subjects: Rheumatism -- Periodicals; 616.723005
• DOI: 10.1136/annrheumdis-2016-eular.2935
• Languages: English
• ISSNs: 0003-4967

Abstract

Abstract : Background: Biologics are very effective to autoimmune diseases including rheumatoid arthritis and ankylosing spondylitis, but could cause infectious diseases as adverse effects. Human T-lymphotropic virus type-I (HTLV-I) is a retrovirus that infects 10 to 20 million people worldwide (1). The virus is associated with severe diseases, such as adult T-cell leukemia/lymphoma and HTLV-I-associated myelopathy (HAM). It is unknown if HTLV-I proliferates when biologics including TNF inhibitors (TNFi) are used. Objectives: We evaluated effects of TNFi to HTLV-I infected cells regarding cytokine profiles, TNF receptor (TNFR), mRNA of associated proteins, proviral load (PVL) and apoptosis. Methods: We used an HTLV-I infected cell line named HCT-5, which is derived from cells in spinal fluid of a patient with HAM. Multiplex cytokine/chemokine bead assays, cytokine levels assessed by enzyme-linked immunosorbent assays (ELISA), fluorescence activated cell sorting (FACS) analysis of TNFR, immunofluorescence staining of GAG, which is a HTLV-1 structural protein, quantitative reverse transcription PCR for Tax (2) and HTLV-I bZIP factor (HBZ) (3), which are needed for immortalization and proliferation of HTLV-I infected cells respectively, HTLV-I PVL (4), and apoptotic ladder detection were performed. We also used Jurkat as a control cell line. Results: Multiplex cytokine/chemokine bead assays showed supernatants of HCT-5 without any stimuli had time-dependent elevation of IL-6, TNF-alpha, RANTES and ICAM-1. Supernatants of HCT-5 treated with infliximab (IFX), adalimumab (ADA), etanercept (ETN), golimumab (GLM) and certolizumab pegol (CZP) assessed by ELISA showed no differences in levels of these molecules compared to that of HCT-5 with phosphate buffered saline (PBS). In FACS analysis, except for TNFR 2 with ETN, expressions of TNFR 1 and TNFR 2 with each TNFi did not differ from PBS. TNFR 2 declined and internalization of TNFR 2 was detected in HCT-5 with ETN. There was no differences in levels of mRNA of Tax (Figure A, PBGD: house keeping gene) and HBZ (Figure B ) with TNFi compared to that with PBS. PVL were also not changed (Figure C, CTR: without any reagents). Immunofluorescence staining of GAG showed no differences between each TNFi and PBS. DNA ladder was not detected in any TNFi and PBS. Conclusions: In vitro, TNFi did not affect the cytokine profiles, TNFR 1, mRNA of associated proteins, proviral load and apoptosis of HCT-5. Only ETN caused internalization of TNFR 2. References: de The G, Bomford R. An HTLV-I vaccine: why, how, for whom? AIDS Res Hum Retroviruses. 1993;9:381–6. Harhaj EW, Harhaj NS. Mechanisms of persistent NF-kappaB activation by HTLV-I tax. IUBMB Life. 2005;57:83–91. Matsuoka M, Jeang KT. Human T-cell leukemia virus type 1 (HTLV-1) and leukemic transformation: viral infectivity, Tax, HBZ and therapy. Oncogene. 2011;30:1379–89. Verdonck K, Gonzalez E, Van Dooren S, Vandamme AM, Vanham G, Gotuzzo E. Human T-lymphotropic virus 1: recent knowledge about an ancient infection. Lancet Infect Dis. 2007;7:266–81. Disclosure of Interest: None declared