Download PDF
Thromb Haemost 2019; 119(03): 377-383
DOI: 10.1055/s-0038-1677032
Cellular Haemostasis and Platelets
Georg Thieme Verlag KG Stuttgart · New York

Novel Murine Model of Immune Thrombocytopaenia through Immunized CD41 Knockout Mice

Xin Li
1   Department of Hematology, Qilu Hospital, Shandong University, Jinan, Shandong, China
,
Shu-wen Wang
1   Department of Hematology, Qilu Hospital, Shandong University, Jinan, Shandong, China
,
Qi Feng
1   Department of Hematology, Qilu Hospital, Shandong University, Jinan, Shandong, China
,
Yu Hou
1   Department of Hematology, Qilu Hospital, Shandong University, Jinan, Shandong, China
2   Shandong Provincial Key Laboratory of Immunohematology, Qilu Hospital, Shandong University, Jinan, Shandong, China
,
Nan Lu
3   Institute of Diagnostics, School of Medicine, Shandong University, Jinan, Shandong, China
,
Chun-hong Ma
4   Department of Immunology, Shandong University School of Medicine, Shandong University, Jinan, Shandong, China
,
Cheng-jiang Gao
4   Department of Immunology, Shandong University School of Medicine, Shandong University, Jinan, Shandong, China
,
Ming Hou
1   Department of Hematology, Qilu Hospital, Shandong University, Jinan, Shandong, China
2   Shandong Provincial Key Laboratory of Immunohematology, Qilu Hospital, Shandong University, Jinan, Shandong, China
5   Leading Research Group of Scientific Innovation, Department of Science and Technology of Shandong Province, Jinan, Shandong, China
,
Jun Peng
1   Department of Hematology, Qilu Hospital, Shandong University, Jinan, Shandong, China
2   Shandong Provincial Key Laboratory of Immunohematology, Qilu Hospital, Shandong University, Jinan, Shandong, China
› Author Affiliations Funding This work was supported by grants from the National Natural Science Foundation of China (No. 91442204, No. 81770133, No. 81370623, No. 81770114, No. 81800112, No. 81500095), Natural Science Foundation of Shandong Province (ZR2017PH022, ZR2017PH041) and Tai Shan Scholar Foundation of Shandong Province.
Further Information

Publication History

22 July 2018

28 November 2018

Publication Date:
10 January 2019 (online)

    Permissions and Reprints

Abstract

Immune thrombocytopaenia (ITP) is the most common autoimmune bleeding disorder, where platelets are destroyed by auto-antibodies and/or cell-mediated mechanisms. To understand the pathogenesis of ITP and explore novel therapeutics, three types of animal models have been used: passive ITP, secondary ITP and platelet-induced ITP. However, the first two are not ideal for chronic ITP pathophysiology where both T cell and B cell play important roles in platelet destruction. The most efficient model to mimic chronic ITP is developed by Chow et al through transferring splenocytes from platelet-immune CD61-knockout (KO) mice into mice with severe combined immunodeficiency (SCID). However, placental defects are evident in 25% of CD61-KO females and post-natal haemorrhage does occur, reducing the survival rate of embryos and pups. Compared with CD61-KO mice, CD41-KO ones do not present such problems. In our study, we employ CD41-KO mice as another source of immunized spleen cells. We evaluated our model with existing standards. Transferred SCID mice presented typical features of ITP, such as reduced platelet counts in the peripheral blood, increased anti-platelet antibody levels in the serum and reduced mature megakaryocytes in the bone marrow. What is more, lymphocyte-depletion experiments showed the role of CD8+ T cells in mature megakaryocyte decrease and thrombocytopaenia. And we confirmed the antibody-mediated platelet destruction by phagocytosis in the spleen. Our study develops another efficient murine ITP model through immunized CD41-KO mice.

Keywords

bleeding disorder - immune thrombocytopaenia - mouse model - CD41 - SCID mouse

Authors' Contributions

J.P. and M.H. designed the research, analysed the data and wrote the paper. X.L. performed the research, analysed the data and wrote the paper. S.-w.W, Q.F. and Y.H. performed the research and analysed the data. N.L., C.-h.M. and C.-j.G. analysed the data.


Supplementary Material

 

  • References

  • 1 Rodeghiero F, Stasi R, Gernsheimer T. , et al. Standardization of terminology, definitions and outcome criteria in immune thrombocytopenic purpura of adults and children: report from an international working group. Blood 2009; 113 (11) 2386-2393
    CrossrefPubMedGoogle Scholar
  • 2 Neunert CE. Current management of immune thrombocytopenia. Hematology (Am Soc Hematol Educ Program) 2013; 2013: 276-282
    CrossrefPubMedGoogle Scholar
  • 3 Provan D, Stasi R, Newland AC. , et al. International consensus report on the investigation and management of primary immune thrombocytopenia. Blood 2010; 115 (02) 168-186
    CrossrefPubMedGoogle Scholar
  • 4 Neunert C, Lim W, Crowther M, Cohen A, Solberg Jr L, Crowther MA. ; American Society of Hematology. The American Society of Hematology 2011 evidence-based practice guideline for immune thrombocytopenia. Blood 2011; 117 (16) 4190-4207
    CrossrefPubMedGoogle Scholar
  • 5 Crow AR, Song S, Semple JW, Freedman J, Lazarus AH. IVIg inhibits reticuloendothelial system function and ameliorates murine passive-immune thrombocytopenia independent of anti-idiotype reactivity. Br J Haematol 2001; 115 (03) 679-686
    CrossrefPubMedGoogle Scholar
  • 6 Katsman Y, Foo AH, Leontyev D, Branch DR. Improved mouse models for the study of treatment modalities for immune-mediated platelet destruction. Transfusion 2010; 50 (06) 1285-1294
    CrossrefPubMedGoogle Scholar
  • 7 Chow L, Aslam R, Speck ER. , et al. A murine model of severe immune thrombocytopenia is induced by antibody- and CD8+ T cell-mediated responses that are differentially sensitive to therapy. Blood 2010; 115 (06) 1247-1253
    CrossrefPubMedGoogle Scholar
  • 8 Aslam R, Hu Y, Gebremeskel S. , et al. Thymic retention of CD4+CD25+FoxP3+ T regulatory cells is associated with their peripheral deficiency and thrombocytopenia in a murine model of immune thrombocytopenia. Blood 2012; 120 (10) 2127-2132
    CrossrefPubMedGoogle Scholar
  • 9 Aslam R, Kapur R, Segel GB. , et al. The spleen dictates platelet destruction, anti-platelet antibody production, and lymphocyte distribution patterns in a murine model of immune thrombocytopenia. Exp Hematol 2016; 44 (10) 924-930
    CrossrefPubMedGoogle Scholar
  • 10 Guo L, Yang L, Speck ER. , et al. Allogeneic platelet transfusions prevent murine T-cell-mediated immune thrombocytopenia. Blood 2014; 123 (03) 422-427
    CrossrefPubMedGoogle Scholar
  • 11 Guo L, Kapur R, Aslam R. , et al. CD20+ B-cell depletion therapy suppresses murine CD8+ T-cell-mediated immune thrombocytopenia. Blood 2016; 127 (06) 735-738
    CrossrefPubMedGoogle Scholar
  • 12 Guo L, Kapur R, Aslam R. , et al. Antiplatelet antibody-induced thrombocytopenia does not correlate with megakaryocyte abnormalities in murine immune thrombocytopenia. Scand J Immunol 2018; 88 (01) e12678
    PubMedGoogle Scholar
  • 13 Kapur R, Catalina MD, Aslam R, Speck ER, Francovitch RF, Semple JW. A highly purified form of staphylococcal protein A alleviates murine immune thrombocytopenia (ITP). Br J Haematol 2018; 183 (03) 501-503
    CrossrefPubMedGoogle Scholar
  • 14 Kapur R, Aslam R, Kim M. , et al. Thymic-derived tolerizing dendritic cells are upregulated in the spleen upon treatment with intravenous immunoglobulin in a murine model of immune thrombocytopenia. Platelets 2017; 28 (05) 521-524
    CrossrefPubMedGoogle Scholar
  • 15 Zufferey A, Speck ER, Machlus KR. , et al. Mature murine megakaryocytes present antigen-MHC class I molecules to T cells and transfer them to platelets. Blood Adv 2017; 1 (20) 1773-1785
    CrossrefPubMedGoogle Scholar
  • 16 Hou Y, Feng Q, Xu M. , et al. High-dose dexamethasone corrects impaired myeloid-derived suppressor cell function via Ets1 in immune thrombocytopenia. Blood 2016; 127 (12) 1587-1597
    CrossrefPubMedGoogle Scholar
  • 17 Lawler J, Weinstein R, Hynes RO. Cell attachment to thrombospondin: the role of ARG-GLY-ASP, calcium, and integrin receptors. J Cell Biol 1988; 107 (6 Pt 1): 2351-2361
    CrossrefPubMedGoogle Scholar
  • 18 Engleman VW, Nickols GA, Ross FP. , et al. A peptidomimetic antagonist of the alpha(v)beta3 integrin inhibits bone resorption in vitro and prevents osteoporosis in vivo. J Clin Invest 1997; 99 (09) 2284-2292
    CrossrefPubMedGoogle Scholar
  • 19 Albelda SM, Mette SA, Elder DE. , et al. Integrin distribution in malignant melanoma: association of the beta 3 subunit with tumor progression. Cancer Res 1990; 50 (20) 6757-6764
    PubMedGoogle Scholar
  • 20 Cheresh DA, Harper JR. Arg-Gly-Asp recognition by a cell adhesion receptor requires its 130-kDa alpha subunit. J Biol Chem 1987; 262 (04) 1434-1437
    CrossrefPubMedGoogle Scholar
  • 21 Hodivala-Dilke KM, McHugh KP, Tsakiris DA. , et al. Beta3-integrin-deficient mice are a model for Glanzmann thrombasthenia showing placental defects and reduced survival. J Clin Invest 1999; 103 (02) 229-238
    CrossrefPubMedGoogle Scholar
  • 22 Reynolds LE, Wyder L, Lively JC. , et al. Enhanced pathological angiogenesis in mice lacking beta3 integrin or beta3 and beta5 integrins. Nat Med 2002; 8 (01) 27-34
    CrossrefPubMedGoogle Scholar
  • 23 Ginsberg MH, Du X, O'Toole TE, Loftus JC. Platelet integrins. Thromb Haemost 1995; 74 (01) 352-359
    Article in Thieme ConnectPubMedGoogle Scholar
  • 24 Phillips DR, Charo IF, Parise LV, Fitzgerald LA. The platelet membrane glycoprotein IIb-IIIa complex. Blood 1988; 71 (04) 831-843
    CrossrefPubMedGoogle Scholar
  • 25 Tronik-Le Roux D, Roullot V, Poujol C, Kortulewski T, Nurden P, Marguerie G. Thrombasthenic mice generated by replacement of the integrin alpha(IIb) gene: demonstration that transcriptional activation of this megakaryocytic locus precedes lineage commitment. Blood 2000; 96 (04) 1399-1408
    CrossrefPubMedGoogle Scholar
  • 26 Zhang J, Varas F, Stadtfeld M, Heck S, Faust N, Graf T. CD41-YFP mice allow in vivo labeling of megakaryocytic cells and reveal a subset of platelets hyperreactive to thrombin stimulation. Exp Hematol 2007; 35 (03) 490-499
    CrossrefPubMedGoogle Scholar
  • 27 Doi T, Homma H, Mezawa S. , et al. Mechanisms for increment of platelet associated IgG and platelet surface IgG and their implications in immune thrombocytopenia associated with chronic viral liver disease. Hepatol Res 2002; 24 (01) 23
    CrossrefPubMedGoogle Scholar
  • 28 McMillan R, Longmire RL, Tavassoli M, Armstrong S, Yelenosky R. In vitro platelet phagocytosis by splenic leukocytes in idiopathic thrombocytopenic purpura. N Engl J Med 1974; 290 (05) 249-251
    CrossrefPubMedGoogle Scholar
  • 29 Kuwana M, Okazaki Y, Kaburaki J, Kawakami Y, Ikeda Y. Spleen is a primary site for activation of platelet-reactive T and B cells in patients with immune thrombocytopenic purpura. J Immunol 2002; 168 (07) 3675-3682
    CrossrefPubMedGoogle Scholar
  • 30 Clarkson SB, Bussel JB, Kimberly RP, Valinsky JE, Nachman RL, Unkeless JC. Treatment of refractory immune thrombocytopenic purpura with an anti-Fc gamma-receptor antibody. N Engl J Med 1986; 314 (19) 1236-1239
    CrossrefPubMedGoogle Scholar
  • 31 Robson HN. Idiopathic thrombocytopenic purpura. Q J Med 1949; 18 (72) 279-297
    PubMedGoogle Scholar
  • 32 Lo E, Deane S. Diagnosis and classification of immune-mediated thrombocytopenia. Autoimmun Rev 2014; 13 (4-5): 577-583
    CrossrefPubMedGoogle Scholar
  • 33 Neschadim A, Branch DR. Mouse models of autoimmune diseases: immune thrombocytopenia. Curr Pharm Des 2015; 21 (18) 2487-2497
    CrossrefPubMedGoogle Scholar
  • 34 Byzova TV, Rabbani R, D'Souza SE, Plow EF. Role of integrin alpha(v)beta3 in vascular biology. Thromb Haemost 1998; 80 (05) 726-734
    Article in Thieme ConnectPubMedGoogle Scholar
  • 35 Leng XH, Hong SY, Larrucea S. , et al. Platelets of female mice are intrinsically more sensitive to agonists than are platelets of males. Arterioscler Thromb Vasc Biol 2004; 24 (02) 376-381
    CrossrefPubMedGoogle Scholar
  • 36 Legorreta-Herrera M, Mosqueda-Romo NA, Nava-Castro KE, Morales-Rodríguez AL, Buendía-González FO, Morales-Montor J. Sex hormones modulate the immune response to Plasmodium berghei ANKA in CBA/Ca mice. Parasitol Res 2015; 114 (07) 2659-2669
    CrossrefPubMedGoogle Scholar
  • 37 Živković I, Bufan B, Petrušić V. , et al. Sexual diergism in antibody response to whole virus trivalent inactivated influenza vaccine in outbred mice. Vaccine 2015; 33 (42) 5546-5552
    CrossrefPubMedGoogle Scholar
  • 38 Živković I, Petrović R, Arsenović-Ranin N. , et al. Sex bias in mouse humoral immune response to influenza vaccine depends on the vaccine type. Biologicals 2018; 52: 18-24
    CrossrefPubMedGoogle Scholar
  • 39 Larcombe AN, Foong RE, Bozanich EM. , et al. Sexual dimorphism in lung function responses to acute influenza A infection. Influenza Other Respir Viruses 2011; 5 (05) 334-342
    CrossrefPubMedGoogle Scholar
  • 40 Laffont S, Rouquié N, Azar P. , et al. X-Chromosome complement and estrogen receptor signaling independently contribute to the enhanced TLR7-mediated IFN-α production of plasmacytoid dendritic cells from women. J Immunol 2014; 193 (11) 5444-5452
    CrossrefPubMedGoogle Scholar
  • 41 Li S, Wang L, Zhao C, Li L, Peng J, Hou M. CD8+ T cells suppress autologous megakaryocyte apoptosis in idiopathic thrombocytopenic purpura. Br J Haematol 2007; 139 (04) 605-611
    CrossrefPubMedGoogle Scholar
  • 42 Kojouri K, Vesely SK, Terrell DR, George JN. Splenectomy for adult patients with idiopathic thrombocytopenic purpura: a systematic review to assess long-term platelet count responses, prediction of response, and surgical complications. Blood 2004; 104 (09) 2623-2634
    CrossrefPubMedGoogle Scholar