Targeting human synovium using homing peptides identified by in vivo phage display

PhDRheumatoid arthritis (RA) is a chronic inflammatory condition affecting diarthrodial synovial joints. Non-random patterns of inflammatory cell recruitment suggest the presence of synovial-specific vascular determinants which enable recruitment of specific inflammatory cell subsets. Using a model whereby human synovial and skin tissue is transplanted into SCID mice, we have previously used in vivo peptide phage display to identify novel peptide sequences which confer synovial homing specificity to human synovium. This synovial localisation was blocked by co-administration of free peptide thus confirming its specificity. In this project the in vivo homing properties of the peptide were further explored. The synovial localization of the synovial-specific phage was shown to be specifically increased after intragraft injection of TNFα. Sequence homology was shown between the expressed CKSTHDRLC (3.1) peptide and an extracellular domain of the leucocyte integrin mac-1. The homing properties of the free peptide were investigated by conjugation to the radioisotopes 111In and 99mTc. No significant differences were found in vivo between homing of the 3.1 monomeric peptide to transplanted human skin and synovium. The influence of valency and size of the molecules were investigated through the development of novel techniques: polymerization of the peptide was achieved by conjugation to radiolabelled streptavidin and fluorescent microspheres. In vivo experiments found no significant difference between localization of polymerised 3.1 or scrambled control peptide to either transplanted skin or synovium with either construct. Despite the negative results reported here, the techniques described have potential for the investigation of other targeted short-peptide sequences. Finally, the model was further developed as a tool for the pre-clinical imaging of human synovium in vivo using an 111In- conjugated anti-E-selectin antibody. It was shown that this could be used to resolve specific from non-specific uptake and hence represents, potentially, a powerful new tool for the development of human tissue-specific targeting strategies

Interleukin-17+CD8+T Cells Are Enriched in the Joints of Patients With Psoriatic Arthritis and Correlate With Disease Activity and Joint Damage Progression

OBJECTIVE: Psoriatic arthritis (PsA) is associated with HLA class I genes, in contrast to the association with HLA class II in rheumatoid arthritis (RA). Since IL-17+ cells are considered important mediators of synovial inflammation, we sought to determine whether IL-17–producing CD8+ T cells may be found in the joints of patients with PsA and whether these cells might contribute to the disease process. METHODS: Mononuclear cells from paired samples of synovial fluid (SF) and peripheral blood (PB) from patients with PsA or patients with RA were stimulated ex vivo, and CD4− T cells were examined by flow cytometry for cytokine expression, cytotoxic markers, and frequencies of γ/δ or mucosal-associated invariant T cells. Clinical measures of arthritis activity (C-reactive protein [CRP] level, erythrocyte sedimentation rate [ESR], Disease Activity Score in 28 joints [DAS28]) and power Doppler ultrasound (PDUS) scores for the presence of active synovitis in the aspirated knee were recorded and assessed for correlations with immunologic markers. RESULTS: Within the CD3+ T cell compartment, both IL-17+CD4− (predominantly CD8+) and IL-17+CD4+ T cells were significantly enhanced in the SF compared to the PB of patients with PsA (P = 0.0003 and P = 0.002, respectively; n = 21), whereas in patients with RA, only IL-17+CD4+ T cells were increased in the SF compared to the PB (P = 0.008; n = 14). The frequency of IL-17+CD4− T cells in PsA SF was positively correlated with the CRP level (r = 0.52, P = 0.01), ESR (r = 0.59, P = 0.004), and DAS28 (r = 0.52, P = 0.01), and was increased in patients with erosive disease (P < 0.05). In addition, the frequency of IL-17+CD4− T cells positively correlated with the PDUS score, a marker for active synovitis (r = 0.49, P = 0.04). CONCLUSION: These results show, for the first time, that the PsA joint, but not the RA joint, is enriched for IL-17+CD8+ T cells. Moreover, the findings reveal that the levels of this T cell subset are correlated with disease activity measures and the radiographic erosion status after 2 years, suggesting a previously unrecognized contribution of these cells to the pathogenesis of PsA

Real world long-term impact of intensive treatment on disease activity, disability and health-related quality of life in rheumatoid arthritis.

Abstract Background The emphasis on treating rheumatoid arthritis (RA) intensively reduces disease activity but its impact in routine care is uncertain. We evaluated temporal changes in disease activities and outcomes in a 10-year prospective observational cohort study of patients in routine care at one unit. Methods The Guy’s and St Thomas’ RA cohort was established in 2005. It involved most RA patients managed in this hospital. Clinical diagnoses of RA were made by rheumatologists. Patients were seen regularly in routine care. Each visit included measurement of disease activity scores for 28 joints (DAS28), health assessment questionnaire scores (HAQ) and EuroQol scores. Patients received intensive treatments targeting DAS28 remission. Results In 1693 RA patients mean DAS28 scores fell from 2005 to 15 by 11% from 4.08 (95% CI: 3.91, 4.25) in 2005 to 3.64 (3.34, 3.78); these falls were highly significant (p  5.1) decreased (25% in 2005; 16% in 2015). In 752 patients seen at least annually for 3 years, persisting remission (68 patients) and intermittent remission (376 patients) were associated with less disability and better health related quality of life. Over time biologic use increased, but they were used infrequently in patients in persistent remission. Conclusions Over 10 years an intensive management strategy in a routine practice setting increased combination DMARD and biologic use: disease activity levels declined; this association is in keeping with a causal relationship. Patients who achieved remission, even transiently, had better functional outcomes than patients never achieving remission