animal models in idiopathic inflammatory myopathies how to overcome a translational roadblock

    Animal models in idiopathic inflammatory myopathies: How to overcome a translational roadblock? Ali Afzali, Tobias Ruck, Heinz Wiendl, Sven G Meuth PII: DOI: Reference: S1568-9972(17)30054-X doi:10.1016/j.autrev.2017.03.001 AUTREV 1980 To appear in: Autoimmunity Reviews Received date: Accepted date: February 2017 13 February 2017 Please cite this article as: Afzali Ali, Ruck Tobias, Wiendl Heinz, Meuth Sven G., Animal models in idiopathic inflammatory myopathies: How to overcome a translational roadblock?, Autoimmunity Reviews (2017), doi:10.1016/j.autrev.2017.03.001 This is a PDF file of an unedited manuscript that has been accepted for publication As a service to our customers we are providing this early version of the manuscript The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain ACCEPTED MANUSCRIPT Animal models in idiopathic inflammatory myopathies: How to overcome a translational roadblock? RI P T Ali Afzali1,*, Tobias Ruck2,*, Heinz Wiendl2 and Sven G Meuth2 Technical University of Munich, Germany Department of Neurology, University of Münster, Germany SC NU * equal contribution MA Corresponding Author: Tobias Ruck, Department of Neurology, University of Münster, Albert-Schweitzer-Campus 1, 48149 Münster, Germany, Tel +49-251-83-46811, PT ED Fax +49-251-83-46812, tobias.ruck@ukmuenster.de Abstract CE Idiopathic inflammatory myopathies (IIMs) encompass a heterogenic group of rare muscle diseases with common symptoms including muscle weakness and the pres- AC ence of certain histological features Since the pathogenesis remains unclear, therapeutic approaches in general comprise unspecific immunosuppression strategies that have been met with limited success Therefore, a deeper understanding of the underlying pathophysiological mechanisms is critically required to assist in development of targeted therapies Animal models have proven to be tremendously helpful in mechanistic studies and allow researchers to overcome the inevitable restrictions of human research Although the number of different IIM models has drastically increased over the last few decades, a model that exhibits the phenotypical and histopathological hallmarks of IIM is still missing Recent publications have shown promising results addressing different pathophysiological issues like mechanisms of onset, chronifica- ACCEPTED MANUSCRIPT tion or relapse in IIM However, a standardization of the methodology is critically required in order to improve comparability and transferability among different groups T Here we provide an overview of the currently available IIM models including our own SC vantages and give perspectives to their future use RI P C-peptide based small-peptide model, critically discuss their advantages and disad- Keywords: Inflammatory myopathies, myositis, animal models, antigens, infections, MA NU transgenic model Abbreviations ED AD, Alzheimer's disease; AMPD, adenosine mono phosphate deaminase; ANA, antinuclear antibodies; ANCA, anticytoplasmatic antibodies; AP-1, activating protein 1; APC, antigen PT presenting cell; APP, amyloid precursor protein; Aβ, amyloid β; BACE-1, β-APP cleaving enzyme 1; BiP, binding immunoglobulin protein; BMDC, bone marrow derived DC; C ele- CE gans, Caenorhabditis elegans; CCL, C-C motif ligand; cdk5, cyclin dependent kinase 5; CFA, Complete Freund's adjuvant; CHIKV, Chikungunya virus; CIM, C-protein induced myositis; AC CK, creatine kinase; CMMM, canine myositis of masticatory muscle; CP2, C-protein fragment 2; CPIM, C-protein peptide induced myositis; CPM, canine polymyositis; CVB, coxsackie virus B; CX3CL, chemokine C-X3-C motif ligand ; CX3CR, chemokine C-X3-C motif receptor; Daf, decay accelerating factor; DC, dendritic cells; DM, dermatomyositis; dPGS, dendritic polyglycerol sulfate; EAE, experimental autoimmune encephalomyelitis; EAM, experimental autoimmune myositis; FOXP3, Forkhead box 3; Grp78, glucose-regulated protein 78; GSK3β, glycogen synthase kinase 3β; HMGCR, 3-hydroxy-3-methylglutaryl-coenzyme A reductase; HRS, histidyl transfer RNA synthetase; HSP, heat shock protein; i.m., intramuscular/ly; i.p., intraperitoneal/ly; i.v., intravenous/ly; ICAM-1, intercellular adhesion molecule 1; ICOS, inducible T cell costimulator; IIM, idiopathic inflammatory myopathies;; IVIG, intravenous immunoglobulin; KFL, Krüppel-like factor; L infantum, Leishmania infantum; LFA-1, lymphocyte ACCEPTED MANUSCRIPT function-associated antigen 1; LPS, lipopolysaccharide; LT, lymphotoxin; Ly-6G, lymphocyte antigen complex; M tuberculosis, Mycobacterium tuberculosis; MAC, membrane attack complex; MB, myosin B fraction; MBL, mannose binding lectin; MCK, muscle creatine kinase; RI P T MCP, monocyte chemotactic protein; mer, monomer; MHC-I, myosin heavy chain I; MIF, migration inhibitory factor; MIP, macrophage inflammatory protein; MLC, myosin light chain; MPS, Methylprednisolone; mTOR, mechanistic target of rapamycin; MyD88, myeloid differ- SC entiation primary response gene 88, NF-κB, nuclear factor kappa-light-chain-enhancer of NU activated B cells; OLE, Oleuropin aglycon; PBMC, peripheral blood mononuclear cells; PKM1/M2, pyruvate kinase M1/M2; PM, polymyositis; poly (I-C), Polyinosinic:polycytidylic MA acid, PS1, preseniline-1; Ptx, pertussis toxin; Rag, recombination activating gene; rec., recombinant; ROS, reactive oxygen species; RRV, Ross River virus; RyR, Ryanodine-receptor; s.c., subcutaneous/ly; sIBM, sporadic Inclusion body myositis; spCIM, small peptide CIM; ED SR, Sarcoplasmatic reticulum; Syt VII, Synaptotagmin VII; ThS, Thioflavin S; TLR, Toll-like PT receptor; TNF-Fc, TNF-α receptor fusion protein; TRE, Tetracycline response element; Treg, regulatory T cell; TRIF, TIR-domain-containing adapter-inducing interferon-β; UPR, unfolded AC CE protein response; VCAM-1, vascular adhesion molecule 1; VLA-4, very late antigen-4 ACCEPTED MANUSCRIPT Introduction Idiopathic inflammatory myopathies (IIMs) comprise a group of relatively rare muscle T diseases (IIMs include dermatomyositis (DM), polymyositis (PM), sporadic inclusion RI P body myositis (sIBM) and necrotizing myositis (NM)) that display heterogeneous clinical phenotypes and occur secondary to systemic disorders such as vasculitides or SC connective tissue disease [1-5] The clinical characteristics of IIMs include progressive muscle weakness that is accompanied by intact sensitivity and tendon reflexes, NU muscle pain and elevated serum creatine kinase (CK) levels [1-3, 6] IIMs can be fur- MA ther distinguished by observing the pattern of affected muscles, patient’s age at disease onset, additional involvement of organs, detected autoantibodies and response to treatment [1, 3, 4] Diagnostic investigations frequently require electromyographic ED measurements and histological evaluation of muscle specimens [4, 7] PT Patients suffering from DM and PM predominantly present symmetric muscle weakness in the proximal parts of the extremities in an either chronically progressive CE or relapsing-remitting disease course [1, 2, 6, 7] DM is further characterized by addi- AC tional skin alterations such as efflorescences, swelling, flush or telangiectasia and the increased coincidence of malignancies Complement-mediated destruction of endomysial blood capillaries with autoantibodies directed against the endothelium is a pathological hallmark of DM In response to antibody binding, an immunological cascade is triggered leading to the formation of membrane attack complexes (MAC) Histological stainings show a predominant presence of CD4+ T and B lymphocytes in affected muscle specimens Phenotypes including capillary necrosis, perivascular damage and loss of muscle fibers also correlate with the inflammatory response [1, 2, 4, 6, 8, 9] The progressive destruction of muscle fibers in PM is assumed to be mainly mediated by cytotoxic CD8+ T cells that attack major histocompatibility complex (MHC) I expressing muscle fibers Muscle biopsies from PM patients show an endo- C.33.44.55.54.78.65.5.43.22.2.4 22.Tai lieu Luan 66.55.77.99 van Luan an.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.22 Do an.Tai lieu Luan van Luan an Do an.Tai lieu Luan van Luan an Do an ACCEPTED MANUSCRIPT mysial infiltration of CD8+ T lymphocytes surrounding necrotic and non-necrotic muscle fibers [1-3, 7, 10] T Sporadic IBM starts by affecting the distal parts of the extremities and is ac- RI P companied by asymmetrically distributed muscle atrophy In contrast to DM and PM, the pathophysiology is instead marked by the coincidence of neuroimmunological SC and neurodegenerative components whereby the causal link still remains unclear Similar to PM, a predominant endomysial accumulation of CD8+ T cells and presence NU of necrotic muscle fibers is seen in immunohistological stainings The neurodegenerative component is manifested by intracellular “rimmed vacuoles” consisting of ag- MA gregated misfolded proteins Interestingly, as seen in Alzheimer’s disease (AD), amyloid-related and hyperphosphorylated tau proteins are detectable in these pathogno- ED monic aggregates [1, 9, 11] Amyloid β (Aβ) has been proposed to be pathophysio- PT logically relevant Aβ is cleaved from the amyloid precursor protein (APP) by the βand γ-secretase and is assumed to be (neuro)toxic In contrast, APP is cleaved under AC 13] CE physiologic conditions by the a-secretase, releasing neuroprotective metabolites [12, Despite the typical characteristics of different IIM subtypes, physicians struggle with the diagnosis due to inconsistent or overlapping histological and serological findings A controversial debate has arisen concerning the diagnostic criteria proposed by Bohan and Peter almost 40 years ago [14] Several adjustments have been made over the last few decades that suggest a common IIM subtypes should be diversified further into smaller entities characterized by specific hallmarks such as circulating antibodies or the presence of certain immunohistological features [15-18] Although the incidence (1:100.000) of IIM is relatively low, affected patients suffer from onerous disabilities limiting quality of life and life expectancy Up to now, causal and selective therapeutic concepts still show limited efficacy and severe side Stt.010.Mssv.BKD002ac.email.ninhd 77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77t@edu.gmail.com.vn.bkc19134.hmu.edu.vn.Stt.010.Mssv.BKD002ac.email.ninhddtt@edu.gmail.com.vn.bkc19134.hmu.edu.vn C.33.44.55.54.78.65.5.43.22.2.4 22.Tai lieu Luan 66.55.77.99 van Luan an.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.22 Do an.Tai lieu Luan van Luan an Do an.Tai lieu Luan van Luan an Do an ACCEPTED MANUSCRIPT effects [9, 19, 20] A better understanding of the underlying pathogenic mechanisms will enable novel therapeutic approaches emphasizing the necessity of intensified T efforts in basic research of IIM Although the usage of animal models in basic re- RI P search has become a polarizing issue in societal debates, their value in the development and risk stratification of promising pharmacological approaches has been inevi- SC tably proven over recent decades [21, 22] Unfortunately, in the context of IIM, the variety and features presented by animal models are currently insufficient and lack NU the histological as well as phenotypic properties of IIM, hindering reproducibility and MA practicability [23, 24] Over the last 20 years, the number of publications referring to IIM has increased introducing different kinds of animal models with certain benefits and limitations (Fig 1A-B) The range of animal models reaches from naturally oc- ED curring myositis to nutritional, transgenic, infectious and immunological models that PT mimic certain features of IIM first described by Wagner and Unverricht over a century ago (Fig 1A, Table and 2) [25, 26] In this review, we will discuss important find- CE ings as well as advantages and disadvantages of different IIM models and provide AC perspectives to improve their relevance for translational research Moreover, we demonstrate data on our own approach to establish a small-peptide mouse model of IIM Stt.010.Mssv.BKD002ac.email.ninhd 77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77t@edu.gmail.com.vn.bkc19134.hmu.edu.vn.Stt.010.Mssv.BKD002ac.email.ninhddtt@edu.gmail.com.vn.bkc19134.hmu.edu.vn C.33.44.55.54.78.65.5.43.22.2.4 22.Tai lieu Luan 66.55.77.99 van Luan an.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.22 Do an.Tai lieu Luan van Luan an Do an.Tai lieu Luan van Luan an Do an ACCEPTED MANUSCRIPT 2.1 Natural myositis in animals Myositis in general is not exclusive to humans and can also occur in different spe- T cies Canine myositis has been investigated over the last 30 years and displays cer- RI P tain similarities to human IIM Up to now, two types of canine myositis have been identified: a locally occurring form known as canine masticatory muscles myositis SC (CMMM) and a general form that symmetrically affects the extremities called canine polymyositis (CPM) Both types feature specific characteristics of IIM such as bilat- NU eral, symmetric generalized muscle atrophy and weakness, cutaneous lesions, elec- MA tromyographic signs of myopathy (e.g positive sharp waves, fibrillation potentials, high frequency discharges) and circulating immune complexes [27, 28] Recently, a member of the myosin binding protein-C family specifically expressed within and on ED the surface of masticatory muscle type 2M fibers was found to be responsible for the PT immune response in CMMM [29] In addition to phenotypical differences, there are also histological differences that can be used to distinguish between CMMM and CE CPM CMMM histologically resembles aspects of DM showing a predominant infiltra- AC tion of both B and CD4+ lymphocytes and an increased expression of MHC-II on the adventitia and endothelium of endo- and perimysial capillaries [30-32] In contrast, CPM shows features of human PM where CD8+ T cells predominantly infiltrate muscle fibers [30, 31] Shelton and colleagues investigated the expression of genes involved in innate and adaptive immunity in both CMMM and CPM specimens through microarray analyses and qPCR experiments [32] Genes involved in macrophage and dendritic cell (DC) activation (CD68, CD40, decorin), migration (CCL4, MCP-2, MMP19), MHC-I (DLA-A, β2-microglobulin) and MHC-II (e.g., DLA-DR, DQ) antigen presentation as well as in B cell growth (ly86), development, migration (CCL 21, CXCL 12) and activation (lck, CD40, CD37) were upregulated in both CMMM and CPM Interestingly, genes regulating molecules of the complement pathway (C1, C1r, Stt.010.Mssv.BKD002ac.email.ninhd 77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77t@edu.gmail.com.vn.bkc19134.hmu.edu.vn.Stt.010.Mssv.BKD002ac.email.ninhddtt@edu.gmail.com.vn.bkc19134.hmu.edu.vn C.33.44.55.54.78.65.5.43.22.2.4 22.Tai lieu Luan 66.55.77.99 van Luan an.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.22 Do an.Tai lieu Luan van Luan an Do an.Tai lieu Luan van Luan an Do an ACCEPTED MANUSCRIPT C1S, C3, C6 and C7) were found to be upregulated only in CMMM specimens Furthermore, genes encoding pro-inflammatory (IL-18, IL-24, IL-13Rα2, IRF-1, T TNFRSF5, caspase 1), anti-inflammatory (TIMP3, IL10R3, IL18BP) or molecules in- RI P volved in tissue remodeling (different collagen subtypes, laminin and TGFBR3) showed increased expression levels [32] SC The alterations in gene and protein expression found in CMMM and CPM specimens demonstrate various similarities to human IIM Canine myositis models NU also have some major advantages in they not require disease induction proce- MA dures and permit study of the disease under naturally occurring conditions [30] It would be interesting to see how CMMM or CPM dogs would respond to established treatment regimens or novel treatment approaches However, experimental settings ED for canine-based research are not established in most laboratories, reagents and an- CE PT tibodies are restricted and animal housing is expensive and time-consuming 2.2 Nutritional myositis models AC Environmental factors are consistently debated as contributing factors in the etiology of IIM Research into neurodegenerative diseases such as AD or sIBM has recently focused more specifically on cholesterol For late onset AD, a diet triggered animal model featuring specific histological hallmarks of AD was established by feeding rabbits a cholesterol enriched diet [33] Chen and colleagues evaluated whether this animal model suitably represents aspects of sIBM by examining histological and molecular-biological features of muscle specimens [34, 35] Muscle sections from treated rabbits showed characteristic histological features similar to those seen in sIBM patients These histological features included infiltration of CD11b+ cells, intramyofibril vacuoles as well as increased intramuscular deposition of Aβ, ubiquitin and hyperphosphorylated tau Although the incidence of these histological findings was relaStt.010.Mssv.BKD002ac.email.ninhd 77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77t@edu.gmail.com.vn.bkc19134.hmu.edu.vn.Stt.010.Mssv.BKD002ac.email.ninhddtt@edu.gmail.com.vn.bkc19134.hmu.edu.vn C.33.44.55.54.78.65.5.43.22.2.4 22.Tai lieu Luan 66.55.77.99 van Luan an.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.22 Do an.Tai lieu Luan van Luan an Do an.Tai lieu Luan van Luan an Do an ACCEPTED MANUSCRIPT [157] W Coley, S Rayavarapu, G.S Pandey, R.L Sabina, J.H Van der Meulen, B Ampong, et al., The molecular basis of skeletal muscle weakness in a mouse model of inflammatory myopathy, Arthritis and rheumatism 64 (2012) 3750-9 T [158] W Coley, S Rayavarapu, J.H van der Meulen, A.S Duba and K Nagaraju, RI P Daily supplementation of D-ribose shows no therapeutic benefits in the MHC-I transgenic mouse model of inflammatory myositis, PloS one (2013) e65970 [159] M Freret, L Drouot, A Obry, S Ahmed-Lacheheb, C Dauly, S Adriouch, et SC al., Overexpression of MHC class I in muscle of lymphocyte-deficient mice causes a severe myopathy with induction of the unfolded protein response, NU Am J Pathol 183 (2013) 893-904 [160] D Figarella-Branger, M Civatte, C Bartoli and J.-F Pellissier, Cytokines, & Nerve 28 (2003) 659-682 MA chemokines, and cell adhesion molecules in inflammatory myopathies, Muscle [161] S Chakrabarti, K.S Kobayashi, R.A Flavell, C.B Marks, K Miyake, D.R ED Liston, et al., Impaired membrane resealing and autoimmune myositis in synaptotagmin VII-deficient mice, The Journal of cell biology 162 (2003) 543- PT [162] A Rodriguez, P Webster, J Ortego and N.W Andrews, Lysosomes behave CE as Ca2+-regulated exocytic vesicles in fibroblasts and epithelial cells, The Journal of cell biology 137 (1997) 93-104 [163] N.A Young, R Sharma, A.K Friedman, B.H Kaffenberger, B Bolon and W.N AC Jarjour, Aberrant muscle antigen exposure in mice is sufficient to cause myositis in a Treg cell-deficient milieu, Arthritis and rheumatism 65 (2013) 3259-70 [164] A Ghirardello, E Borella, M Beggio, F Franceschini, M Fredi and A Doria, Myositis autoantibodies and clinical phenotypes, Auto- immunity highlights (2014) 69-75 [165] H Gunawardena, The Clinical Features of Myositis-Associated Autoantibodies: a Review, Clinical Reviews in Allergy & Immunology 52 (2017) 45-57 [166] M Satoh, S Tanaka, A Ceribelli, S.J Calise and E.K.L Chan, A Comprehensive Overview on Myositis-Specific Antibodies: New and Old Biomarkers in Idiopathic Inflammatory Myopathy, Clinical Reviews in Allergy & Immunology 52 (2017) 1-19 Stt.010.Mssv.BKD002ac.email.ninhd 77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77t@edu.gmail.com.vn.bkc19134.hmu.edu.vn.Stt.010.Mssv.BKD002ac.email.ninhddtt@edu.gmail.com.vn.bkc19134.hmu.edu.vn C.33.44.55.54.78.65.5.43.22.2.4 22.Tai lieu Luan 66.55.77.99 van Luan an.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.22 Do an.Tai lieu Luan van Luan an Do an.Tai lieu Luan van Luan an Do an ACCEPTED MANUSCRIPT [167] J.-P Simon, I Marie, F Jouen, O Boyer and J Martinet, Autoimmune Myopathies: Where Do We Stand?, Frontiers in Immunology (2016) [168] M Labrador-Horrillo, M.A Martínez, A Selva-O'Callaghan, E Trallero- T Araguás, J.M Grau-Junyent, M Vilardell-Tarrés, et al., Identification of a novel RI P myositis-associated antibody directed against cortactin, Autoimmunity Reviews 13 (2014) 1008-1012 [169] P Cherin, C Belizna, O Cartry, G Lascu-Dubos, C de Jaeger, J.-C Delain, SC et al., Long-term subcutaneous immunoglobulin use in inflammatory myopathies: A retrospective review of 19 cases, Autoimmunity Reviews 15 NU (2016) 281-286 [170] A Rongvaux, T Willinger, J Martinek, T Strowig, S.V Gearty, L.L MA Teichmann, et al., Development and function of human innate immune cells in a humanized mouse model, Nature Biotechnology 32 (2014) 364-372 [171] L.D Shultz, F Ishikawa and D.L Greiner, Humanized mice in translational ED biomedical research, Nature Reviews Immunology (2007) 118-130 [172] L.D Shultz, M.A Brehm, J.V Garcia-Martinez and D.L Greiner, Humanized PT mice for immune system investigation: progress, promise and challenges, AC CE Nature reviews Immunology 12 (2012) 786-98 Stt.010.Mssv.BKD002ac.email.ninhd 77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77t@edu.gmail.com.vn.bkc19134.hmu.edu.vn.Stt.010.Mssv.BKD002ac.email.ninhddtt@edu.gmail.com.vn.bkc19134.hmu.edu.vn C.33.44.55.54.78.65.5.43.22.2.4 22.Tai lieu Luan 66.55.77.99 van Luan an.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.22 Do an.Tai lieu Luan van Luan an Do an.Tai lieu Luan van Luan an Do an ACCEPTED MANUSCRIPT Tables Table Animal models for sIBM Transgenic C57BL/6 & MCK-APP/PS1 mice Chronic myopathy with mononuclear cell infiltration and vacuoles filled with Aβ42 and ThS, clinical signs present C57BL/6 & MA Transgenic Chronic myositis with CD11b+ cells and intracellular deposits with Aβ and hyperphosphorylated Tau, low incidence, other tissues affected NU SJL/J mice Diet with 2% cholesterol RI P Rabbit Characteristics SC Diettriggered Protocol T Induction Animals MLC-APP mice ED SJL/J mice HSA-LTab mice PT Transgenic Not mentioned CE Transgenic Nematode Human Aβ C overexpressed elegans in muscle Chronic myopathy with absent infiltrates, intracellular Aβ42 and ThS deposits in type 2M fibers, clinical signs present Chronic myopathy with immune cell infiltration, MHC-I upregulation, intracellular aggregates and clinical signs of myopathy Publications Chen et al 2008 Liu et al 2016 Sugarman et al 2001, Sugarman et al 2005, Kitazawa et al 2006 Moussa et al 2006, Shtifmann et al 2010 Bauer et al 2015 Chronic myopathy with immuno- Link et al.1995, reactivity to ThS and Aβ, no Reboliedo et immunological participation, al 2008 clinical signs present AC Table Animal models for DM and PM Induction Animals Protocol Characteristics Publications Hargis 1985, Hargis 1988 Shelton et al 2006 Natural Dogs Canine Polymyositis Chronic myositis CD8+ T cells with moderate, symmetric affection of extremities; genetic alterations of innate and adaptive immunity similar to PM Infectious BALB/c, Swiss C3H, COH mice Coxsackievirus B1 Acute progressive, selfsustaining myositis, T cells involved in chronification, high death rate, low incidence in certain strains Infectious CD-1, C57BL/6 mice Ray et al 1979 Strongwater 1984, Yitterberg et al 1987 Ross River virus, Mild acute myositis, unspecific Morrison et Chikungunya with arthritis and tendosynovitis, al 2006, virus complement mediated Morrison et al 2011 Stt.010.Mssv.BKD002ac.email.ninhd 77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77t@edu.gmail.com.vn.bkc19134.hmu.edu.vn.Stt.010.Mssv.BKD002ac.email.ninhddtt@edu.gmail.com.vn.bkc19134.hmu.edu.vn C.33.44.55.54.78.65.5.43.22.2.4 22.Tai lieu Luan 66.55.77.99 van Luan an.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.22 Do an.Tai lieu Luan van Luan an Do an.Tai lieu Luan van Luan an Do an ACCEPTED MANUSCRIPT Publications Induction Animals Protocol Characteristics Infectious CBA/J mice Trypanosoma cruzi Mild acute myositis and myocarditis driven by CD8+ T cells Infectious Syrian hamsters Leishmania Infantum Mild acute myositis with CD8+ T Paciello 2010 cells Purified Myosin Immunolog- Lewis rats, ical C57BL/6 mice C-Protein fragment T RI P SC Kojima et al 1997, Suzuki et al 2005, Scuderi et al 2006 Moderate, remitting myositis Kohyama et with high incidence driven by T al 1999, cell and a complex reaction with Sugihara et chemokines, cytokines and al 2007 costimulatory molecules Moderate, self-limiting myositis Okiyama et similar to active CIM al 2012 ED Transfer of CIM splenocytes cocultured with naïve BMDCs preincubated with CP2 and IL2, simultaneous s.c CFA injection Immunolog- Wistar rats Laminin Mild myositis without clinical ical signs, endomysial infiltrates with macrophages and lymphocytes Immunolog- C57BL/6, Amino terminal Moderate chronic, focal myoical B6.G7, portion of HRS sitis with CD4+ and CD8+ infilNod.Idd3/5 without CFA trates lasting for weeks, inmice duction independent of TLR2, TLR4 and of mature T cells AC CE PT Immunolog- C57BL/6 ical Matsubara et al 1987, Ito et al 2002, Rosenberg et al 1987 MA Immunolog- Lewis rats, ical SJL/J, BALB/C, C57BL/6 mice Mild, self-limiting myositis without clinical signs, a complex immune reaction with predominance of macrophages and expression of adhesion molecules Moderate, self-limiting EAM with a complex reaction comprising lymphocytes, chemokines and cytokines, restricted to SJL/J mice NU Immunolog- Guinea Myosin B fraction ical pigs, Lewis rats, SJL/J mice Sun 1993, Andersson et al 2003 Transgenic Transgenic C57BL/6 (H+T+ mice) C57BL/6 (Syt VII-/-) MHC-I Synaptotagmin VII Chronic myopathy driven by cell stress mechanisms, no T cell infiltration Chronic myositis with clinical signs and endomysial infiltrates consisting of CD3+, CD11b+ and Ly-6G+ cells, ANA positive Stt.010.Mssv.BKD002ac.email.ninhd 77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77t@edu.gmail.com.vn.bkc19134.hmu.edu.vn.Stt.010.Mssv.BKD002ac.email.ninhddtt@edu.gmail.com.vn.bkc19134.hmu.edu.vn Nakano et al 2005 Sohejima 2011, Harlow 2012 Nagaraju et al 2000, Li et al 2009 Chakrabati et al 2003 C.33.44.55.54.78.65.5.43.22.2.4 22.Tai lieu Luan 66.55.77.99 van Luan an.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.22 Do an.Tai lieu Luan van Luan an Do an.Tai lieu Luan van Luan an Do an ACCEPTED MANUSCRIPT Table Agents used for treatment Model Agent Application Characteristics Binderit Twice a day from day Ameliorated disease course EAM MB Treg infusion day before immunization Ameliorated disease course EAM Anti-CX3CL-1 mAb I.p injections times a week during time of immunization EAM I.v MPS EAM IVIG EAM Tacrolimus EAM Am80 RI P T RRV Myositis Sugihara et al 2007 NU SC I.v for days days after immunization I.v twice per week during immunization MA IVIG CIM Anti-ICOS mAb CIM Anti-IL6R mAb Allenbach et al 2009 Szuzuki et al 2005 ED CIM Rulli et al 2009 Decreased endomysial infiltration accompanied by reduced mRNA levels of TNF-α, IFN-γ and perforin Twice during 12 h Higher rates of apoptotic after last immun- T cells ization I.v for days Less severe histological scores I.v day 14-35 Less severe histological scores, ICAM-1 decreased Preventive: s.c No influence on inciinjections on day dence and number of 1-21 necrotic fibers, reduction of infiltrating cells, IL-4, IL-10 and IFN-γ increased, MCP-1 and RANTES decreased Therapeutic: s.c No influence on inciinjections on day dence and number of 15-28 necrotic fibers, reduced histological scores Preventive: Oral Muscle strength inday before 1st creased, histological immunization until scores reduced, lympho10 days after last penia, Tregs increased, immunization effector T cell proliferation reduced Therapeutic: Oral Muscle strength in1 day after last creased, histological immunization until scores reduced, lympho10 days penia, Tregs increased PT CE Rapamycin AC EAM Publication Preventive: I.p from day Therapeutic: I.p from day Attenuated disease course and histological scores Ameliorated CIM, CD11b+ and CD8+ T cells reduced, reduced IL-1a and MCP-1 Reduced incidence and severity Reduced incidence and severity Stt.010.Mssv.BKD002ac.email.ninhd 77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77t@edu.gmail.com.vn.bkc19134.hmu.edu.vn.Stt.010.Mssv.BKD002ac.email.ninhddtt@edu.gmail.com.vn.bkc19134.hmu.edu.vn Schneider et al 2000 Wada et al 2001 Nemoto et al 2001 Ohyanagi et al 2009 Ohyanagi et al 2009 Prevel et al 2013 Prevel et al 2013 Katsumata et al 2007 Okiyama et al 2009 Okiyama et al 2009 C.33.44.55.54.78.65.5.43.22.2.4 22.Tai lieu Luan 66.55.77.99 van Luan an.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.22 Do an.Tai lieu Luan van Luan an Do an.Tai lieu Luan van Luan an Do an ACCEPTED MANUSCRIPT Agent Application Characteristics Publication CIM Anti-CD25 mAb (data not shown) No difference CIM IL-1R antagonist Continuous s.c Less severe histological scores Okiyama et al 2012 Sugihara et al 2012 CIM Anti-IL-1R mAb I.p times week- Less severe histological ly scores Sugihara et al 2012 CIM TNFR-Fc CIM Anti-TNFa mAb I.p times weekly I.p times weekly Sugihara et al 2012 Sugihara et al 2012 CIM dPGS RI P T Model SC Less severe histological scores Less severe histological scores ED Transgenic C elegans Fed with OLE (12.5–500 µM, 100 µl/plate) Cu2+ MA Transgenic Oleuropin aglyC elegans (OLE) NU S.c for 11 days Impaired severity of CIM Oishi et al days after immun2014 ization Injected i.p for weeks (2 injections per week) Diomede et al 2013 Increased number of amyloid deposits, Less Aβ oligomers, loco motor impairment attenuated Attenuated disease courses and comparable decreased Grp78 levels Minniti et al 2009, Aldunate et al 2012 Rayavarapu et al 2013 AC CE PT H+T+ mice Bortezomib Less Aβ aggregates and less paralysis Stt.010.Mssv.BKD002ac.email.ninhd 77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77t@edu.gmail.com.vn.bkc19134.hmu.edu.vn.Stt.010.Mssv.BKD002ac.email.ninhddtt@edu.gmail.com.vn.bkc19134.hmu.edu.vn C.33.44.55.54.78.65.5.43.22.2.4 22.Tai lieu Luan 66.55.77.99 van Luan an.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.22 Do an.Tai lieu Luan van Luan an Do an.Tai lieu Luan van Luan an Do an ACCEPTED MANUSCRIPT Table Intracellular signaling pathways in animal models of IIM Signaling Model Animals Effect Publications pathway SJL/J mice Calcineurin  EAM BALB/c KLF-2  HRSinduced myositis C57BL/6, B6.G7, NOD.Idd3/5, DO11.10/Rag2-/-, C3H7Hej MCK-APP C57BL/6 C57BL/6 Chaperones  HSP  PT CE AC : upregulation Chaperones  HSP UPR with GRP78, BiP and caspase 12  NF-κB  : downregulation Sojima et al 2011 RI P Involved in hyperphosphorylated tau deposition Kitazawa et al 2006 Cell stress activation Drake et al 2003, Fonte et al 2002, Link et al 1999 Cell stress activation Nagaraju et al 2005, Rayavarapu et al 2013 MA H+T+ Howard et al 2002, ED C elegans Involved in activation of innate immune system with TLR  NU MyDD88  GSK-3β  C.elegans myositis Nemoto et al 2001 Prevel et al 2013 SC TRIF  Cdk5  Disease course +, ICAM-1  Disease course +, T cell proliferation  T EAM Apoptosis induction  Immune response  +: amelioration Stt.010.Mssv.BKD002ac.email.ninhd 77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77t@edu.gmail.com.vn.bkc19134.hmu.edu.vn.Stt.010.Mssv.BKD002ac.email.ninhddtt@edu.gmail.com.vn.bkc19134.hmu.edu.vn C.33.44.55.54.78.65.5.43.22.2.4 22.Tai lieu Luan 66.55.77.99 van Luan an.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.22 Do an.Tai lieu Luan van Luan an Do an.Tai lieu Luan van Luan an Do an ACCEPTED MANUSCRIPT Figure Legends Figure History of animal models for IIM (A) Timeline of milestones for the es- T tablishment of animal models in IIM (B) The number of publications on myositis RI P models since its first description SC Figure Targets of myositis models (A) Schematic overview of skeletal muscle structure and currently used targets in animal models of IIM For greater clarity, scale MA NU and colors were adjusted Figure Muscle inflammation score (MIS) Histological grading based on the ED number of necrotic muscle fibers found in single muscle block (A) Grade 0, muscle tissue without muscle fiber necrosis (B) Grade 1, a lesion with up to necrot- PT ic muscle fibers (C) Grade 2, 5-30 muscle fibers involved (D) Grade 3, involvement of more than 30 muscle fibers (E) Grade 4, diffuse, extensive lesions 0.5 point was AC block [101] CE added, when the same grade was found in multiple lesions within a single muscle Figure Histological stainings of C-protein-peptide immunized C57BL/6 mice Quadriceps femoris muscle from immunized and sham-immunized (PBS) C57BL/6 mice were isolated and prepared for H.E stainings (n = 3) A histological score (A) on the basis of the amount of necrotic and invaded muscle fibers was established and used for the analysis of H.E stainings (B) Representative stainings of muscle tissue from mice immunized with indicated peptides 10, 12 and 13 (P10, P12, P13) Wildtype muscle tissue from C57BL/6 mice (“native”-line) and sham-immunized mice (“PBS”-line) were used as controls The average scores of peptide-immunized mice are depicted as “mean value line” (MV-line) All data are shown as the mean SEM Stt.010.Mssv.BKD002ac.email.ninhd 77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77t@edu.gmail.com.vn.bkc19134.hmu.edu.vn.Stt.010.Mssv.BKD002ac.email.ninhddtt@edu.gmail.com.vn.bkc19134.hmu.edu.vn C.33.44.55.54.78.65.5.43.22.2.4 22.Tai lieu Luan 66.55.77.99 van Luan an.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.22 Do an.Tai lieu Luan van Luan an Do an.Tai lieu Luan van Luan an Do an ACCEPTED MANUSCRIPT Figure Immunological readouts of C-protein-peptide immunized C57BL/6 mice Immune cells were isolated from inguinal lymph nodes of immunized C57BL/6 T mice and used for further readouts (n = 3) (A) Immune cells were seeded on a 96 RI P well plate and stimulated with the particular peptide and CD3 (1 μg/ml) for 48 h Supernatants were taken and analyzed for IL-2 concentrations by ELISA (R&D sys- SC tems) Immune cells without peptide restimulation and from sham-immunized C57BL/6 mice served as controls (B) The proliferation of immune cells was as- NU sessed by measuring ATP release upon stimulation with CD3 (1 μg/ml) and the particular peptide or PBS respectively with the ATPLite Luminescence Assay System MA (PerkinElmer) according to the manufacturer’s instructions All data are shown as the ED mean SEM P < 0.05 was considered as significant * PT Figure Multiple immunizations with peptide 13 (KDGVELTREDSFKARYRF) (A) Three consecutive immunizations with peptide 13 in ascending order from the CE foot pads to the tail and to the back led to severe spCIM disease course with reduc- AC tion of body weight gain and (B) diminished test time and speed in Rotarod-Test (C) Macroscopic signs of muscle inflammation and destruction: enlarged lymph nodes and fatty tissue reorganization of the muscle (D) Lymphocytes isolated from dissected lymph nodes showed a tendency of increased activation with increased levels of CD25, CD40L and CD69 assessed by flow cytometry All data are shown as the mean SEM P < 0.05 was considered as significant Stt.010.Mssv.BKD002ac.email.ninhd 77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77t@edu.gmail.com.vn.bkc19134.hmu.edu.vn.Stt.010.Mssv.BKD002ac.email.ninhddtt@edu.gmail.com.vn.bkc19134.hmu.edu.vn C.33.44.55.54.78.65.5.43.22.2.4 22.Tai lieu Luan 66.55.77.99 van Luan an.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.22 Do an.Tai lieu Luan van Luan an Do an.Tai lieu Luan van Luan an Do an Figure AC CE PT ED MA NU SC RI P T ACCEPTED MANUSCRIPT Stt.010.Mssv.BKD002ac.email.ninhd 77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77t@edu.gmail.com.vn.bkc19134.hmu.edu.vn.Stt.010.Mssv.BKD002ac.email.ninhddtt@edu.gmail.com.vn.bkc19134.hmu.edu.vn C.33.44.55.54.78.65.5.43.22.2.4 22.Tai lieu Luan 66.55.77.99 van Luan an.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.22 Do an.Tai lieu Luan van Luan an Do an.Tai lieu Luan van Luan an Do an PT ED MA NU SC RI P T ACCEPTED MANUSCRIPT AC CE Figure Stt.010.Mssv.BKD002ac.email.ninhd 77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77t@edu.gmail.com.vn.bkc19134.hmu.edu.vn.Stt.010.Mssv.BKD002ac.email.ninhddtt@edu.gmail.com.vn.bkc19134.hmu.edu.vn C.33.44.55.54.78.65.5.43.22.2.4 22.Tai lieu Luan 66.55.77.99 van Luan an.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.22 Do an.Tai lieu Luan van Luan an Do an.Tai lieu Luan van Luan an Do an AC CE PT ED MA NU SC RI P T ACCEPTED MANUSCRIPT Figure Stt.010.Mssv.BKD002ac.email.ninhd 77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77t@edu.gmail.com.vn.bkc19134.hmu.edu.vn.Stt.010.Mssv.BKD002ac.email.ninhddtt@edu.gmail.com.vn.bkc19134.hmu.edu.vn C.33.44.55.54.78.65.5.43.22.2.4 22.Tai lieu Luan 66.55.77.99 van Luan an.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.22 Do an.Tai lieu Luan van Luan an Do an.Tai lieu Luan van Luan an Do an AC CE PT ED MA NU SC RI P T ACCEPTED MANUSCRIPT Figure Stt.010.Mssv.BKD002ac.email.ninhd 77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77t@edu.gmail.com.vn.bkc19134.hmu.edu.vn.Stt.010.Mssv.BKD002ac.email.ninhddtt@edu.gmail.com.vn.bkc19134.hmu.edu.vn C.33.44.55.54.78.65.5.43.22.2.4 22.Tai lieu Luan 66.55.77.99 van Luan an.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.22 Do an.Tai lieu Luan van Luan an Do an.Tai lieu Luan van Luan an Do an AC CE PT ED MA NU SC RI P T ACCEPTED MANUSCRIPT Figure Stt.010.Mssv.BKD002ac.email.ninhd 77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77t@edu.gmail.com.vn.bkc19134.hmu.edu.vn.Stt.010.Mssv.BKD002ac.email.ninhddtt@edu.gmail.com.vn.bkc19134.hmu.edu.vn C.33.44.55.54.78.65.5.43.22.2.4 22.Tai lieu Luan 66.55.77.99 van Luan an.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.C.33.44.55.54.78.655.43.22.2.4.55.22 Do an.Tai lieu Luan van Luan an Do an.Tai lieu Luan van Luan an Do an AC Figure CE PT ED MA NU SC RI P T ACCEPTED MANUSCRIPT Stt.010.Mssv.BKD002ac.email.ninhd 77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77.77.99.44.45.67.22.55.77.C.37.99.44.45.67.22.55.77t@edu.gmail.com.vn.bkc19134.hmu.edu.vn.Stt.010.Mssv.BKD002ac.email.ninhddtt@edu.gmail.com.vn.bkc19134.hmu.edu.vn