SAT0010 ANTI-CD30 IMMUNOTHERAPY AMELIORATES BONE AND CARTILAGE DESTRUCTION IN EXPERIMENTAL MODEL OF RHEUMATOID ARTHRITIS IN MICE

Abstract

Background:CD30 is a member of the TNF-receptor family and commonly expressed on lymphocytes of Hodgkin lymphoma and anaplastic large cell lymphoma. It has been reported that levels of soluble CD30 in serum and joint fluid is significantly elevated in rheumatoid arthritis (RA). Although RA patients may develop lymphoproliferative disorders (LPD) as a result of immunosuppression by MTX or bDMARDs, safety medications after the regression of LPD for RA have not yet been established.Objectives:To explore the potential of CD30 targeting therapy for RA.Methods:(1) Immuno-histological staining of CD30 was performed for fresh synovial tissues of RA and osteoarthritis (OA). In addition, double immunofluorescence staining of CD30 with CD3, CD20, CD68, CD138 were performed on RA synovial tissue. (2) Brentuximab vedotin (BV) is an anti-CD30 antibody conjugated with monomethyl auristatin E, designed to induce apoptosis of CD30 expressing cells. A multiple myeloma cell line (RPMI8226) was used as a non-lymphoma cell line and plasma cell-like cell line. Immuno-cytological staining for CD30 was performed on RPMI8226. Cells were cultured and harvested on days 0, 1, and 3 to evaluate the effects of BV (50 μl / ml per well). Cytospin specimens were stained by May-Grunwald-Giemsa (MGG) staining for cell counting and by FIFC-terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining for detection of apoptosis. (3) Collagen antibody induced arthritis (CAIA) was induced in DBA/1 mice by arthritogenic cocktail of monoclonal antibodies against type II collagen. BV was administered to the treatment groups (30mg/kg and 70mg/kg n=4 each) and evaluated clinical score, histological findings and levels of SAA, IL-6, and TNFα in serum by ELISA. Student’st-test (two-tailed) was used to determine statistical significance for analysis of synovial tissues and cell line assay. Two way ANOVA with Dunnett’s post hoc analysis was used for multiple comparisons of mice model.Results:(1) The number of CD30-positive cells was significantly higher in RA synovial tissue than in OA synovial tissue (p<0.01) (Fig. 1). CD30-positive cells were detected around the lymphoid follicles. Double immunofluorescence showed CD30 and CD138 double-positive cells in the synovial tissue of RA, suggesting CD30 is predominantly expressed by plasma cells. (2) RPMI8226 cells expressed CD30. BV caused apoptosis of RPMI8226 cells, and the number of cells treated with BV decreased to 95% compared to controls. (3) All control mice (n=4) developed severe arthritis, and their scores reached a peak (score: 13.3) on day 10. In the mice of treatment group of 30 mg/kg, paw swelling was slightly decreased, their clinical score reached a peak (score: 9.3) on day 10. In contrast, paw swelling was significantly reduced in the 70 mg/kg treatment group. The peak of the clinical score was 4.3 on day 10 (Fig.2). Histological score evaluated synovitis with infiltration of inflammatory cells, pannus formation, and erosion of bone and cartilage. Histological score of hind paws were 3.0 ± 0.8 for the control group, 2.7 ± 1.0 for 30 mg/kg group, and 0.7 ± 1.1 for 70 mg/kg group (p<0.01), respectively. The serum levels of SAA and IL-6 of treatment group were lower than those of no treatment group (p<0.01).Conclusion:We showed the expression of CD30 on synovial tissue of RA and the expression of CD30 on plasma cells. In addition, the current study provides the first evidence that BV depletion of CD30-positive cells suppressed arthritis and osteochondral destruction in CAIA mice. Our results may provide an important clue for the development of an effective treatment for RA with iatrogenic immunodeficiency-related LPD.Disclosure of Interests:Minami Matsuhashi: None declared, Keiichiro Nishida Grant/research support from: K. Nishida has received scholarship donation from CHUGAI PHARMACEUTICAL Co., Eisai Co., Mitsubishi Tanabe Pharma and AbbVie GK., Speakers bureau: K. Nishida has received speaking fees from CHUGAI PHARMACEUTICAL Co., Eli Lilly, Janssen Pharmaceutical K.K., Eisai Co. and AYUMI Pharmaceutical Corporation., Yoshihisa Nasu: None declared, Ryuichi Nakahara: None declared, Masahito Watanabe: None declared, Yoshifumi Hotta: None declared, Toshifumi Ozaki: None declared