Parasitology Research

, Volume 117, Issue 8, pp 2623–2633 | Cite as

Brain proteomic differences between wild-type and CD44- mice induced by chronic Toxoplasma gondii infection

  • Jing Yang
  • Fen Du
  • Xiaoliu Zhou
  • Lixia Wang
  • Senyang Li
  • Rui Fang
  • Junlong Zhao
Original Paper


Chronic clinical Toxoplasma gondii (T. gondii) infection is the primary disease state that causes severe encephalitis. CD44 is a member of the cell adhesion molecule family and plays an important role in T. gondii infection. However, proteomic changes in CD44 during chronic T. gondii infection have rarely been reported. Thus, an iTRAQ-based proteomic study coupled with 2D-LC-MS/MS analysis was performed to screen CD44-related proteins during chronic T. gondii infection. As a result, a total of 2612 proteins were reliably identified and quantified. Subsequently, 259, 106, and 249 differentially expressed proteins (DEPs) were compared between CD44- mice (A) vs wild-type mice (B), B vs wild-type mice infected with T. gondii (C), and C vs CD44- mice infected with T. gondii (D). Gene ontology, KEGG pathway, and protein-protein interaction analyses were performed on the DEPs. According to the results, immune-related proteins were altered significantly among the A vs B, B vs C, and C vs D comparisons, which might indicate that chronic T.  gondii infection caused changes in the host immune response. Additionally, Ca2+- and metabolism-related proteins were upregulated in C vs D, which supported the hypothesis that CD44 mediated the production of host Ca2+ and IFN-γ and that the parasite preferentially invaded cells expressing high levels of CD44. The present findings validate and enable a more comprehensive knowledge of the role of CD44 in hosts chronically infected with T. gondii, thus providing new ideas for future studies on the specific functions of CD44 in latent toxoplasmosis.


Toxoplasma gondii CD44- iTRAQ Proteomic Differentially expressed protein (DEP) 



This work was supported by the National Natural Science Foundation of China (R.F., No. 31572510), the Natural Science Foundation of Hubei Province (Grant No. 2017CFA020), and Da Bei Nong Group Promoted Project for Young Scholar of HZAU (Grant No. 2017DBN001).

Compliance with ethical standards

Competing interests

The authors declare that they have no competing interests.

Supplementary material

436_2018_5954_MOESM1_ESM.pdf (33 kb)
Table S1 Mouse proteins identified at critical false discovery rates (Critical FDR < 0.01). (PDF 33 kb)
436_2018_5954_MOESM2_ESM.xls (2.5 mb)
Table S2 List of 2612 trusted proteins identified and quantified in this study. (XLS 2572 kb)
436_2018_5954_MOESM3_ESM.xls (137 kb)
Table S3 List of differentially expressed proteins (DEPs) between CD44- and wild-type mice (fold change > 2 or fold change < 0.5). (XLS 137 kb)
436_2018_5954_MOESM4_ESM.xls (66 kb)
Table S4 List of differentially expressed proteins (DEPs) between wild-type mice and wild-type mice infected with T. gondii (fold change > 2 or fold change < 0.5). (XLS 66 kb)
436_2018_5954_MOESM5_ESM.xls (130 kb)
Table S5 List of differentially expressed proteins (DEPs) between wild-type mice infected with T. gondii and CD44- mice infected with T. gondii(fold change > 2 or fold change < 0.5). (XLS 129 kb)


  1. Alstergren P, Zhu BQ, Glougauer M, Mak TW, Ellen RP, Sodek J (2004) Polarization and directed migration of murine neutrophils is dependent on cell surface expression of CD44. Cell Immunol 231:146–157. CrossRefPubMedGoogle Scholar
  2. Bautista JM, Marin-Garcia P, Diez A, Azcarate IG, Puyet A (2014) Malaria proteomics: insights into the parasite-host interactions in the pathogenic space. J Proteome 97:107–125. CrossRefGoogle Scholar
  3. Blass SL, Pure E, Hunter CA (2001) A role for CD44 in the production of IFN-gamma and immunopathology during infection with Toxoplasma gondii. J Immunol 166:5726–5732CrossRefPubMedGoogle Scholar
  4. Blume M, Nitzsche R, Sternberg U, Gerlic M, Masters SL, Gupta N, McConville MJ (2015) A Toxoplasma gondii gluconeogenic enzyme contributes to robust central carbon metabolism and is essential for replication and virulence. Cell Host Microbe 18:210–220. CrossRefPubMedGoogle Scholar
  5. Brouns MR, Matheson SF, Hu KQ, Delalle I, Caviness VS, Silver J, Bronson RT, Settleman J (2000) The adhesion signaling molecule p190 RhoGAP is required for morphogenetic processes in neural development. Development 127:4891–4903PubMedGoogle Scholar
  6. Collazo CM, Miller C, Yap G, Hieny S, Caspar P, Schwartz RH, Sher A (2000) Host resistance and immune deviation in pigeon cytochrome c T-cell receptor transgenic mice infected with Toxoplasma gondii. Infect Immun 68:2713–2719CrossRefPubMedPubMedCentralGoogle Scholar
  7. Doskaya M et al (2014) Diagnostic value of a Rec-ELISA using Toxoplasma gondii recombinant SporoSAG, BAG1, and GRA1 proteins in murine models infected orally with tissue cysts and oocysts. PLoS One 9:e108329. CrossRefPubMedPubMedCentralGoogle Scholar
  8. Egan ES, Jiang RHY, Moechtar MA, Barteneva NS, Weekes MP, Nobre LV, Gygi SP, Paulo JA, Frantzreb C, Tani Y, Takahashi J, Watanabe S, Goldberg J, Paul AS, Brugnara C, Root DE, Wiegand RC, Doench JG, Duraisingh MT (2015) Malaria. A forward genetic screen identifies erythrocyte CD55 as essential for Plasmodium falciparum invasion. Science 348:711–714. CrossRefPubMedPubMedCentralGoogle Scholar
  9. Foger N, Marhaba R, Zoller M (2000) CD44 supports T cell proliferation and apoptosis by apposition of protein kinases. Eur J Immunol 30:2888–2899.;2888::AID-IMMU2888&#62;3.0.CO;2–4Google Scholar
  10. Freitas do Rosario AP, Lamb T, Spence P, Stephens R, Lang A, Roers A, Muller W, O'Garra A, Langhorne J (2012) IL-27 promotes IL-10 production by effector Th1 CD4+ T cells: a critical mechanism for protection from severe immunopathology during malaria infection. J Immunol 188:1178–1190. CrossRefPubMedGoogle Scholar
  11. Gee K, Lim W, Ma W, Nandan D, Diaz-Mitoma F, Kozlowski M, Kumar A (2002) Differential regulation of CD44 expression by lipopolysaccharide (LPS) and TNF-alpha in human monocytic cells: distinct involvement of c-Jun N-terminal kinase in LPS-induced CD44 expression. J Immunol 169:5660–5672CrossRefPubMedGoogle Scholar
  12. Gitau EN, Kokwaro GO, Newton CRJC, Ward SA (2011) Global proteomic analysis of plasma from mice infected with Plasmodium berghei ANKA using two dimensional gel electrophoresis and matrix assisted laser desorption ionization-time of flight mass spectrometry. Malar J 10:Artn 205. CrossRefGoogle Scholar
  13. Hayashi T, Unno A, Baba M, Ohno T, Kitoh K, Takashima Y (2014) CD44 mediated Hyaluronan adhesion of Toxoplasma gondii-infected leukocytes. Parasitol Int 63:479–484. CrossRefPubMedGoogle Scholar
  14. Henry SC, Traver M, Daniell X, Indaram M, Oliver T, Taylor GA (2010) Regulation of macrophage motility by Irgm1. J Leukoc Biol 87:333–343. CrossRefPubMedGoogle Scholar
  15. Jurzynski C, Gysin J, Pouvelle B (2007) CD44, a signal receptor for the inhibition of the cytoadhesion of CD36-binding Plasmodium falciparum-infected erythrocytes by CSA-binding infected erythrocytes. Microbes Infect 9:1463–1470. CrossRefPubMedGoogle Scholar
  16. Krishnamurthy S, Konstantinou EK, Young LH, Gold DA, Saeij JPJ (2017) The human immune response to Toxoplasma: autophagy versus cell death. PLoS Pathog 13:e1006176. CrossRefPubMedPubMedCentralGoogle Scholar
  17. Li C, Wu Y, Orian-Rousseau V, Zhang Y, Gulbins E, Grassme H (2017) Regulation of Staphylococcus aureus infection of macrophages by CD44, reactive oxygen species and acid sphingomyelinase. Antioxid Redox Signal.
  18. Ling XB, Cohen H, Jin J, Lau I, Schilling J (2009) FDR made easy in differential feature discovery and correlation analyses. Bioinformatics (Oxford, England) 25:1461–1462. CrossRefGoogle Scholar
  19. Londono DP, Alvarez JI, Trujillo J, Jaramillo MM, Restrepo BI (2002) The inflammatory cell infiltrates in porcine cysticercosis: immunohistochemical analysis during various stages of infection. Vet Parasitol 109:249–259CrossRefPubMedGoogle Scholar
  20. Lv L, Wang Y, Feng W, Hernandez JA, Huang W, Zheng Y, Zhou X, Lv S, Chen Y, Yuan ZG (2017) iTRAQ-based differential proteomic analysis in Mongolian gerbil brains chronically infected with Toxoplasma gondii. J Proteome 160:74–83. CrossRefGoogle Scholar
  21. Mahmoudvand H, Saedi Dezaki E, Soleimani S, Baneshi MR, Kheirandish F, Ezatpour B, Zia-Ali N (2015) Seroprevalence and risk factors of Toxoplasma gondii infection among healthy blood donors in south-east of Iran. Parasite Immunol 37:362–367. CrossRefPubMedGoogle Scholar
  22. Masek KS, Fiore J, Leitges M, Yan SF, Freedman BD, Hunter CA (2006) Host cell Ca2+ and protein kinase C regulate innate recognition of Toxoplasma gondii. J Cell Sci 119:4789–4789. CrossRefGoogle Scholar
  23. Mercier A, Ajzenberg D, Devillard S, Demar MP, de Thoisy B, Bonnabau H, Collinet F, Boukhari R, Blanchet D, Simon S, Carme B, Dardé ML (2011) Human impact on genetic diversity of Toxoplasma gondii: example of the anthropized environment from French Guiana. Infect Genet Evol 11:1378–1387. CrossRefPubMedGoogle Scholar
  24. Miwa T, Nagata T, Kojima H, Sekine S, Okumura T (2017) Isoform switch of CD44 induces different chemotactic and tumorigenic ability in gallbladder cancer. Int J Oncol 51:771–780. CrossRefPubMedPubMedCentralGoogle Scholar
  25. Nicolle C, Manceaux LH (2009) On a leishman body infection (or related organisms) of the gondi. 1908. Int J Parasitol 39:863–864CrossRefPubMedGoogle Scholar
  26. Nitzsche R, Zagoriy V, Lucius R, Gupta N (2016) Metabolic cooperation of glucose and glutamine is essential for the lytic cycle of obligate intracellular parasite Toxoplasma gondii. J Biol Chem 291:126–141. CrossRefPubMedGoogle Scholar
  27. Ohshima J, Lee Y, Sasai M, Saitoh T, Su Ma J, Kamiyama N, Matsuura Y, Pann-Ghill S, Hayashi M, Ebisu S, Takeda K, Akira S, Yamamoto M (2014) Role of mouse and human autophagy proteins in IFN-gamma-induced cell-autonomous responses against Toxoplasma gondii. J Immunol 192:3328–3335. CrossRefPubMedGoogle Scholar
  28. Ondriska F, Catar G, Vozarova G (2003) The significance of complement fixation test in clinical diagnosis of toxoplasmosis. Bratisl Lek Listy 104:189–196PubMedGoogle Scholar
  29. Pappas G, Roussos N, Falagas ME (2009) Toxoplasmosis snapshots: global status of Toxoplasma gondii seroprevalence and implications for pregnancy and congenital toxoplasmosis. Int J Parasitol 39:1385–1394. CrossRefPubMedGoogle Scholar
  30. Singleton PA, Bourguignon LY (2004) CD44 interaction with ankyrin and IP3 receptor in lipid rafts promotes hyaluronan-mediated Ca2+ signaling leading to nitric oxide production and endothelial cell adhesion and proliferation. Exp Cell Res 295:102–118. CrossRefPubMedGoogle Scholar
  31. Smith NL, Abdallah DSA, Butcher BA, Denkers EY, Baird B, Holowka D (2013) Toxoplasma gondii inhibits mast cell degranulation by suppressing phospholipase C gamma-mediated Ca2+ mobilization. Front Microbiol 4:Artn 179. CrossRefGoogle Scholar
  32. Stephens R, Langhorne J (2010) Effector memory Th1 CD4 T cells are maintained in a mouse model of chronic malaria. PLoS Pathog 6:e1001208. CrossRefPubMedPubMedCentralGoogle Scholar
  33. Unno A, Kitoh K, Takashima Y (2010) Up-regulation of hyaluronan receptors in Toxoplasma gondii-infected monocytic cells. Biochem Biophys Res Commun 391:477–480. CrossRefPubMedGoogle Scholar
  34. Vuorio J, Vattulainen I, Martinez-Seara H (2017) Atomistic fingerprint of hyaluronan-CD44 binding. PLoS Comput Biol 13:e1005663. CrossRefPubMedPubMedCentralGoogle Scholar
  35. Wang LQ, Dai Y, Qi SW, Sun BD, Wen JL, Zhang L, Tu ZG (2012) Comparative proteome analysis of peripheral blood mononuclear cells in systemic lupus erythematosus with iTRAQ quantitative proteomics. Rheumatol Int 32:585–593. CrossRefPubMedGoogle Scholar
  36. Wang ZX, Zhou CX, Elsheikha HM, He S, Zhou DH, Zhu XQ (2017) Proteomic differences between developmental stages of Toxoplasma gondii revealed by iTRAQ-based quantitative proteomics. Front Microbiol 8:Artn 985. CrossRefGoogle Scholar
  37. Wilson RA (2012) Proteomics at the schistosome-mammalian host interface: any prospects for diagnostics or vaccines? Parasitology 139:1178–1194. CrossRefPubMedGoogle Scholar
  38. Wong NKY, Lai JCY, Maeshima N, Johnson P (2011) CD44-mediated elongated T cell spreading requires Pyk2 activation by Src family kinases, extracellular calcium, phospholipase C and phosphatidylinositol-3 kinase. Cell Signal 23:812–819. CrossRefPubMedGoogle Scholar
  39. Zhao Y, Ferguson DJP, Wilson DC, Howard JC, Sibley LD, Yap GS (2009) Virulent Toxoplasma gondii evade immunity-related GTPase-mediated parasite vacuole disruption within primed macrophages. J Immunol 182:3775–3781. CrossRefPubMedPubMedCentralGoogle Scholar
  40. Zhou DH, Zhao FR, Huang SY, Xu MJ, Song HQ, Su C, Zhu XQ (2013) Changes in the proteomic profiles of mouse brain after infection with cyst-forming Toxoplasma gondii. Parasit Vectors 6:96. CrossRefPubMedPubMedCentralGoogle Scholar
  41. Zimmermann S, Murray PJ, Heeg K, Dalpke AH (2006) Induction of suppressor of cytokine signaling-1 by Toxoplasma gondii contributes to immune evasion in macrophages by blocking IFN-gamma signaling. J Immunol 176:1840–1847CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.State Key Laboratory of Agricultural Microbiology, College of Veterinary MedicineHuazhong Agricultural UniversityWuhanPeople’s Republic of China
  2. 2.Hubei Centre for Animal Diseases Control and PreventionWuhanPeople’s Republic of China
  3. 3.Hubei Provincial Centre for Diseases Control and PreventionWuhanPeople’s Republic of China

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