title: Prothymosin alpha lacking the nuclear localization signal as an effective gene therapeutic strategy in collagen-induced arthritis.
abstract: Prothymosin alpha (ProT) is regulated by c-Myc, an oncoprotein overexpressed in synovium of rheumatoid arthritis, and is associated with cell proliferation. However, ProT also exerts immunomodulatory activities. The growth-promoting activity of ProT can be abolished by deleting its nuclear localization signal (NLS). In this study, we showed that AdProTDeltaNLS, an adenoviral vector encoding ProT lacking the NLS, did not enhance the proliferation of synovial fibroblasts. AdProTDeltaNLS treatment abolished the up-regulation of the MIP-1alpha promoter activity induced by TNF-alpha in synovial fibroblasts. AdProTDeltaNLS suppressed macrophage chemotaxis and reduced macrophage infiltration into the ankle joints in rats with collagen-induced arthritis (CIA). Neutralization test confirmed the involvement of MIP-1alpha in macrophage chemotaxis. Administration of AdProTDeltaNLS reduced the severity of CIA in the clinical, radiographic, and histological aspects. The levels of TNF-alpha (mean +/- SEM, 1261.9 +/- 107.9 vs 2880.1 +/- 561.4 pg/mg total protein; p < 0.05), IL-1beta (56.8 +/- 8.0 vs 109.2 +/- 4.9 pg/mg total protein; p < 0.01), and MIP-1alpha (41.7 +/- 3.6 vs 55.2 +/- 1.1 pg/mg total protein; p < 0.05) in the ankle joints were lower in the AdProTDeltaNLS-treated rats with CIA than those in their control counterparts. In the AdProTDeltaNLS-treated ankle joints, matrix metalloproteinase-9 expression was decreased by 40% and infiltrating macrophages reduced by 50%. Our results demonstrate that intra-articular delivery of AdProTDeltaNLS significantly ameliorated the clinical course of CIA in rats. This study is the first to suggest that ProT lacking the NLS may have therapeutic potential for the management of rheumatoid arthritis.
authors: Department of Microbiology and Immunology, National Cheng Kung University Medical College, 1 Dashiue Road, Tainan 701, Taiwan.
authors: Ai-Li
authors: AL
authors: Shiau
authors: Shih-Yao
authors: SY
authors: Chen
authors: Meng-Ya
authors: MY
authors: Chang
authors: Chih-Hau
authors: CH
authors: Su
authors: Shih-Ye
authors: SY
authors: Chung
authors: Yi-Te
authors: YT
authors: Yo
authors: Chrong-Reen
authors: CR
authors: Wang
authors: Chao-Liang
authors: CL
authors: Wu
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000843:Ankle Joint / Q000276:immunology / Q000473:pathology; D001172:Arthritis, Rheumatoid / Q000139:chemically induced / Q000473:pathology / Q000628:therapy*; D049109:Cell Proliferation; D054405:Chemokine CCL3; D054407:Chemokine CCL4; D002633:Chemotaxis; D003094:Collagen / Q000633:toxicity; D005347:Fibroblasts; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D006801:Humans; D053583:Interleukin-1beta / Q000378:metabolism; D019402:Macrophage Inflammatory Proteins / Q000378:metabolism; D008264:Macrophages / Q000276:immunology; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D019913:Nuclear Localization Signals / Q000032:analysis / Q000235:genetics*; D011498:Protein Precursors / Q000032:analysis / Q000235:genetics*; D051381:Rats; D017384:Sequence Deletion; D013582:Synovial Fluid / Q000737:chemistry; D013947:Thymosin / Q000031:analogs & derivatives* / Q000032:analysis / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D015854:Up-Regulation
chemical_list: D054405:Chemokine CCL3; D054407:Chemokine CCL4; D053583:Interleukin-1beta; D019402:Macrophage Inflammatory Proteins; D019913:Nuclear Localization Signals; D011498:Protein Precursors; D014409:Tumor Necrosis Factor-alpha; C040782:prothymosin alpha; D013947:Thymosin; D003094:Collagen; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9