Journal: Biochemistry and Biophysics Reports

Article Title: IL-38: A new factor in rheumatoid arthritis

doi: 10.1016/j.bbrep.2015.10.015

Figure Lengend Snippet: Characteristics of human IL-38. (A) Western blotting analysis using anti-human IL-38 mAb (H127C) showed that human IL-38 cDNA-transfected 293T cells expressed approximately 18 kDa IL-38 protein in the cells and supernatants. Line 1: Cell lysate of vector-transfected 293T cells. Line 2: Cell lysate of human IL-38 cDNA-transfected-transfected 293T cells. Line 3: Supernatant of vector-transfected 293T cells. Line 4: Supernatant of human IL-38 cDNA-transfected-transfected 293T cells. As control, recombinant human IL-38 (rhIL-38) protein with His tag at C-terminal was used. (B) Standard curve of human IL-38 sandwich ELISA using anti-human IL-38 mAbs (H127C and H160A). (C) Caspase-1, chymase, and PR3 can cleave recombinant human pro-IL-1β (hpro-IL-1β), but not rhIL-38 protein. Western blotting analysis was performed using rabbit anti-human pro-IL-1β polyclonal Ab (Santa Cruz Biotechnology, Dallas, TX) and rabbit anti-human IL-38 polyclonal Ab (established at our laboratory).

Article Snippet: Digestion of recombinant human IL-38 and pro-IL-1β protein (Sino Biological Inc., Beijing, China) by recombinant caspase 1, chymase, and PR3 (Sigma, St. Louis, MO, USA) was performed as reported previously , .

Techniques: Western Blot, Transfection, Plasmid Preparation, Recombinant, Sandwich ELISA

Journal: Biochemistry and Biophysics Reports

Article Title: IL-38: A new factor in rheumatoid arthritis

doi: 10.1016/j.bbrep.2015.10.015

Figure Lengend Snippet: IL-38 expression in RA serum and synovium. (A) Serum levels of IL-38 in RA, OA patients and healthy donors. Serum levels of IL-38 was examined by IL-38 ELISA. Twenty-one of 137 RA (15.3%), one of 26 OA patients (3.9%) and 5 of 56 controls (8.9%) were elevated above the detection limit. Detection limit is 9.35 ng/ml. (B) IL-38 is expressed in RA, but not OA synovium. Synovial tissues were obtained from 7 RA patients and 2 OA patients. Synovial tissues were immunostained with anti-human IL-38 mAb [H127C, mouse IgG2b] (b, c, d) or isotype-matched control mouse IgG2b Ab (a). Representative sections of synovial tissues from 2 RA patients (b, c) and a patient with OA (d) are shown. Original magnification: ×200.

Article Snippet: Digestion of recombinant human IL-38 and pro-IL-1β protein (Sino Biological Inc., Beijing, China) by recombinant caspase 1, chymase, and PR3 (Sigma, St. Louis, MO, USA) was performed as reported previously , .

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Biochemistry and Biophysics Reports

Article Title: IL-38: A new factor in rheumatoid arthritis

doi: 10.1016/j.bbrep.2015.10.015

Figure Lengend Snippet: IL-38 gene deficiency enhances joint inflammation in RA mouse model. Arthritis was initiated in female IL-38 −/− mice and control B6 WT mice via intraperitoneal administration of K/BxN mouse serum. (A) IL-38 mRNA in ankle joints (8 ankles/group from two separate experiments) was determined before or 8 days after K/BxN serum transfer. * p <0.05, normal vs. inflamed joints. (B) Clinical score of K/BxN serum transfer arthritis in WT and IL-38 −/− mice on a 0–16 scale ( n =5/group), * p <0.05, ** p <0.01, WT vs. IL-38 −/− mice. Data are representative of at least 2 separate experiments. (C) Histomorphometric quantification of arthritic tissue. Data were pooled in two independent experiments (10 ankles/group from two separate experiments). * p <0.05, WT vs. IL-38 −/− mice. (D) Histopathologic findings in the ankle joints of representative WT (left) and IL-38 −/− mice (right). Hematoxylin and eosin stained; original magnification ×40. (E) Cytokine mRNA in the ankle joints (10 ankles/group from two separate experiments) was examined at day 7 or 8 arthritis. * p <0.05, WT vs. IL-38 −/− mice. Values in A–C and E are the mean±SEM.

Article Snippet: Digestion of recombinant human IL-38 and pro-IL-1β protein (Sino Biological Inc., Beijing, China) by recombinant caspase 1, chymase, and PR3 (Sigma, St. Louis, MO, USA) was performed as reported previously , .

Techniques: Staining