POS0623 CYTOKINE PRODUCTION BY BLOOD LYMPHOCYTES DEFINES A PROFILE ASSOCIATED WITH NON-REMISSION IN PATIENTS WITH RHEUMATOID ARTHRITIS TREATED WITH TNF INHIBITORS
Authors: B. Hernández-Breijo, E. Rodríguez-Martín, C. García-Hoz, V. Navarro-Compán, C. Sobrino, A. Martínez-Feito, I. Nieto-Gañán, J. Bachiller-Corral, P. Lapuente-Suanzes, G. Bonilla, C. Pijoán-Moratalla, M. Vázquez, A. Balsa, D. Pascual-Salcedo, L. M. Villar, C. Plasencia
Journal: Annals of the Rheumatic Diseases (2021)

Background: In clinical practice no more than 50% of the patients treated with TNF inhibitors (TNFi) achieve remission (REM). Previous investigations suggested that peripheral blood mononuclear cells (PBMC) may be markers associated with the TNFi treatment success 1 . Objectives: This study aims to analyse the intracellular cytokine production by PBMC and its association with REM achievement after 6 months (m) of TNFi treatment in patients with RA. Methods: This was a prospective study including 62 patients with RA starting the 1 st TNFi. PBMC were isolated from patients at baseline and after 6m of treatment with TNFi and cryopreserved until studied. In vitro stimulation and intracellular cytokine production by PBMC was performed as follow: in the presence of 2µg/mL brefeldin and 2µmol/L monensin monocytes were stimulated with 20ng/mL LPS during 4h whereas lymphocytes were stimulated with 50ng/mL phorbol 12-myristate 13-acetate and 750ng/mL ionomycin for 4h at 37°C. To identify IL10-producing B cells, PBMC were pre-incubated with 3µg/mL of CpG oligonucleotide during 20h at 37°C prior to stimulation. Intracellular cytokine production (TNFα, IL6, GM-CSF, IL10) by the different cell subsets (monocytes, CD4 + and CD8 + T cells, naïve and memory B cells) was analysed by flow-cytometry. Clinical activity at baseline and after 6m was assessed by DAS28-ESR. REM was defined as DAS28≤2.6 at 6m. The association between cytokine production by each PBMC subset and REM was analysed through univariable and multivariable logistic regression models. Receiving operating curve (ROC) analysis was used to select the optimal ratio of cytokine production associated with REM status. Results: After 6m of TNFi treatment, 30 (48%) patients achieved REM. No significant differences between REM and non-REM groups were observed for patients’ characteristics at baseline except for DAS28, which was lower in the REM group (non-REM: 5.4±0.9; REM: 4.3±0.9; p&lt;0.0001) (Table 1). Therefore, further analyses were adjusted by baseline DAS28. A lower ratio between calculated with the IL10 and TNFα production by B cells and by CD4 + T cells (IL10 B/TNF CD4) at 6m was found for non-REM patients (non-REM: 0.31 vs REM: 0.54; p=0.007). Based on a ROC analysis, we found that a (IL10 B/TNF CD4)&lt;0.54 at 6 m was significantly associated with a higher probability of non-REM at 6 months (OR: 5.0; 95% CI: 1.1-21.7) (Figure 1). Table 1. Baseline predictors of reduction of disease activity at 12 months from start of abatacept. Linear regression. Baseline patients’ characteristics Total patients (n=62 ) DAS28&gt;2.6 (n=32; 52% ) DAS28≤2.6 (n=30; 48% ) p-value Age (years ) 53±12 53±13 52±10 0.8 Female 55 (89) 30 (94) 25 (83) 0.2 Disease duration (years ) 8 (4-11) 8 (4-12) 7 (3-11) 0.7 RF positive 49 (79) 23 (72) 26 (87) 0.1 ACPA positive 54 (87) 26 (81) 28 (93) 0.2 Smoking habit (n=55 ) 0.2 Non-smokers 26 (47) 16 (55) 10 (38) Smoker 29 (53) 13 (45) 16 (51) Body mass index (kg/m 2 ) 25.9±5.6 25.8±5.7 26.0±5.6 0.9 DAS28 4.9±1.0 5.4±0.9 4.3±0.9 &lt;0.0001 Concomitant csDMARDs 60 (97) 32 (100) 28 (93) 0.3 MTX [±OD] 46 (74) 26 (81) 20 (67) 0.3 Only OD 14 (23) 6 (19) 8 (26) 0.3 Prednisone 36 (58) 19 (59) 17 (57) 0.9 Conclusion: Our results show that the proinflammatory IL10 B/TNF CD4 ratio is associated with non-REM status. It could be useful to analyse the success of TNFi treatment in patients with RA. References: [1]Rodríguez-Martín E, et al. Front Immunol. 2020; 11: 1913. Acknowledgements: ISCIII (PI16/00474; PI16/01092) Disclosure of Interests: Borja Hernández-Breijo: None declared, Eulalia Rodríguez-Martín: None declared, Carlota García-Hoz: None declared, Victoria Navarro-Compán Speakers bureau: Abbvie, Janssen, Lilly, MSD, Novartis, Pfizer and UCB, Grant/research support from: Abbvie, Janssen, Lilly, MSD, Novartis, Pfizer and UCB, Cristina Sobrino: None declared, ANA MARTÍNEZ-FEITO: None declared, Israel Nieto-Gañán: None declared, Javier Bachiller-Corral Speakers bureau: Abbvie, MSD, BMS and Roche, Grant/research support from: Pfizer, Paloma Lapuente-Suanzes: None declared, Gemma Bonilla: None declared, Cristina Pijoán-Moratalla: None declared, Mónica Vázquez: None declared, Alejandro Balsa Speakers bureau: Abbvie, BMS, Nordic, Novartis, Pfizer, Sandoz, Sanofi, Roche and UCB, DORA PASCUAL-SALCEDO: None declared, Luisa María Villar: None declared, Chamaida Plasencia Speakers bureau: AbbVie, Lilly, Novartis, Pfizer, Sanofi, Biogen and UCB