Short Peptides of Innate Immunity Protein Tag7 Inhibit the Production of Cytokines in CFA-Induced Arthritis

Abstract

The pathogenesis of autoimmune arthritis is a hot topic in current research. The main focus of this work was to study cytokines released in CFA-induced arthritis in ICR mice as well as the regulation of blood levels of cytokines by two peptides of the innate immunity protein Tag7 (PGLYRP1) capable of blocking the activation of the TNFR1 receptor. Arthritis was induced by local periarticular single-dose injections of 40 µL of complete Freund's adjuvant (CFA) into the left ankle joints of mice. The levels of chemokines and cytokines in plasma were measured using a Bio-Plex Pro Mouse Cytokine Kit at 3, 10, and 21 days after arthritis induction. Tag7 peptides were shown to decrease the blood levels of the pro-inflammatory cytokines IL-6, TNF, and IL-1β. Administration of peptides also decreased the levels of chemokines MGSA/CXCL1, MIP-2α/CXCL2, ENA78/CXCL5, MIG/CXCL9, IP-10/CXCL10, MCP-1/CCL2, and RANTES/CCL5. Furthermore, a decrease in the levels of cytokines IL7, G-CSF, and M-CSF was demonstrated. Addition of the studied peptides strongly affected IFN-γ concentration. We believe that a decrease in the levels of cytokine IFN-γ was associated with a therapeutic effect of Tag7 peptides manifested in alleviation of the destruction of cartilage and bone tissues in the CFA-induced arthritis.

Keywords: 17.1 and 17.1a peptides; CFA; TNFα; Tag7; arthritis; inflammation; mice.

Figure 1

Changes in the level of pro-inflammatory cytokines ((A) IL-1b; (B) IL-6; (C) TNF) in the plasma 145 of mice after CFA-induced arthritis. (Control—control animals without CFA injection, CFA—animals with induced via CFA arthritis, + saline—animals were treated by phosphate buffered saline [PBS], +17.1a, +17.1, +Norocarp—animals were treated by a specified agent; n = 7 for each group). (p-value: * < 0.05).

Figure 2

Changes in the level of chemokines ((A) CXCL1; (B) CXCL2; (C) CXCL5; (D) CXCL9; (E) CXCL10) in the plasma of mice after CFA-induced arthritis. (Control—control animals without CFA injection, CFA—animals with induced via CFA arthritis, +saline—animals were treated by phosphate buffered saline (PBS), +17.1a, +17.1, +Norocarp—animals were treated by a specified agent, n = 7 for each group). (p-value: * < 0.05).

Figure 3

Changes in the level of CCL2 chemokine (CCL2) in the plasma of mice after CFA212 induced arthritis. (Control—control animals without CFA injection, CFA—animals with induced via CFA arthritis, +saline—animals were treated by phosphate buffered saline (PBS), +17.1a, +17.1, +Norocarp—animals were treated by a specified agent, n = 7 for each group). (p-value: * < 0.05).

Figure 4

Changes in the level of hematopoietic cytokines ((A) IL-7) and growth factors ((B) G-CSF; ((C) M-CSF) in the plasma of mice after CFA-induced arthritis. (Control—control animals without CFA injection, CFA—animals with induced via CFA arthritis, +saline—animals were treated by phosphate buffered saline (PBS), +17.1a, +17.1, +Norocarp—animals were treated by a specified agent, n = 7 for each group). (p-value: * < 0.05).

Figure 5

Changes of the level of cytokines in the effector phase of immune response (IFN-γ) in the plasma of mice after CFA-induced arthritis. (Control—control animals without CFA injection, CFA—animals with induced via CFA arthritis, +saline—animals were treated by phosphate buffered saline (PBS), +17.1a, +17.1, +Norocarp—animals were treated by a specified agent, n = 7 for each group). (p-value: * < 0.05).