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Production and characterization of monoclonal antibodies against Toxoplasma gondii ROP18 with strain-specific reactivity

Published online by Cambridge University Press:  12 February 2020

Famin Zhang ,
Rui Su ,
Chengjian Han ,
Yang Wang ,
Jingyang Li ,
Jinjin Zhu ,
Qingli Luo ,
Lingzhi Chen ,
Junling Zhang  and
Xiaojuan Ding
...Show all authors
Famin Zhang
Affiliation:
Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Provincial Laboratory of Zoonoses of High Institutions, School of Basic Medical Sciences, Anhui Medical University, Hefei, China
Rui Su
Affiliation:
Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Provincial Laboratory of Zoonoses of High Institutions, School of Basic Medical Sciences, Anhui Medical University, Hefei, China
Chengjian Han
Affiliation:
Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Provincial Laboratory of Zoonoses of High Institutions, School of Basic Medical Sciences, Anhui Medical University, Hefei, China
Yang Wang
Affiliation:
Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Provincial Laboratory of Zoonoses of High Institutions, School of Basic Medical Sciences, Anhui Medical University, Hefei, China
Jingyang Li
Affiliation:
Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Provincial Laboratory of Zoonoses of High Institutions, School of Basic Medical Sciences, Anhui Medical University, Hefei, China The Clinical Laboratory of the Third People's Hospital of Heifei, Hefei, China
Jinjin Zhu
Affiliation:
Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Provincial Laboratory of Zoonoses of High Institutions, School of Basic Medical Sciences, Anhui Medical University, Hefei, China
Qingli Luo
Affiliation:
Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Provincial Laboratory of Zoonoses of High Institutions, School of Basic Medical Sciences, Anhui Medical University, Hefei, China
Lingzhi Chen
Affiliation:
Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Provincial Laboratory of Zoonoses of High Institutions, School of Basic Medical Sciences, Anhui Medical University, Hefei, China
Junling Zhang
Affiliation:
Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Provincial Laboratory of Zoonoses of High Institutions, School of Basic Medical Sciences, Anhui Medical University, Hefei, China
Xiaojuan Ding
Affiliation:
Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Provincial Laboratory of Zoonoses of High Institutions, School of Basic Medical Sciences, Anhui Medical University, Hefei, China
Jian Du
Affiliation:
Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Provincial Laboratory of Zoonoses of High Institutions, School of Basic Medical Sciences, Anhui Medical University, Hefei, China Department of Biochemistry and Molecular Biology, Anhui Medical University, Hefei, China
Deyong Chu
Affiliation:
Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Provincial Laboratory of Zoonoses of High Institutions, School of Basic Medical Sciences, Anhui Medical University, Hefei, China
Yihong Cai
Affiliation:
Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Provincial Laboratory of Zoonoses of High Institutions, School of Basic Medical Sciences, Anhui Medical University, Hefei, China Department of Health Inspection and Quarantine, School of Public Health, Anhui Medical University, Hefei, China
Jilong Shen
Affiliation:
Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Provincial Laboratory of Zoonoses of High Institutions, School of Basic Medical Sciences, Anhui Medical University, Hefei, China
Li Yu*
Affiliation:
Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Provincial Laboratory of Zoonoses of High Institutions, School of Basic Medical Sciences, Anhui Medical University, Hefei, China
*
Author for correspondence: Li Yu, E-mail: [email protected]
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Abstract

The rhoptry kinase 18 of Toxoplasma gondii (TgROP18) has been identified as a key virulence factor that allows the parasite to escape from host immune defences and promotes its proliferation in host cells. Although much research is focused on the interaction between host cells and TgROP18, the development of monoclonal antibodies (mAbs) against TgROP18 has not been reported till date. To produce mAbs targeting TgROP18, two hybridomas secreting mAbs against TgROP18, designated as A1 and T2, were generated using cell fusion technology. The subtypes of the A1 and T2 mAbs were identified as IgG3 λ and IgM κ, and peptide scanning revealed that the core sequences of the antigenic epitopes were 180LRAQRRRSELVFE192 and 351NYFLLMMRAEADM363, respectively. The T2 mAb specifically reacted with both T. gondii type I and Chinese I, but not with T. gondii type II, Plasmodium falciparum or Schistosoma japonicum. Finally, the sequences of heavy chain and light chain complementarity-determining regions of T2 were amplified, cloned and characterized, making the modification of the mAb feasible in the future. The development of mAbs against TgROP18 would aid the investigation of the molecular mechanisms underlying the modulation of host cellular functions by TgROP18, and in the development of strategies to diagnose and treat Toxoplasmosis.

Keywords

Antigenic epitopeApicomplexacomplementarity-determining regionsrhoptryTgROP18Toxoplasma gondiitoxoplasmosis
Type
Research Article
Information
Parasitology , Volume 147 , Issue 9 , August 2020 , pp. 940 - 948
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Copyright
Copyright © The Author(s), 2020. Published by Cambridge University Press

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Footnotes

†

These authors contributed equally to this work.

References

Boothroyd, JC (2009) Toxoplasma gondii: 25 years and 25 major advances for the field. International Journal for Parasitology 39, 935–946.CrossRefGoogle ScholarPubMed
Boothroyd, JC and Dubremetz, JF (2008) Kiss and spit: the dual roles of Toxoplasma rhoptries. Nature Reviews: Microbiology 6, 79–88.Google ScholarPubMed
Bradley, PJ and Sibley, LD (2007) Rhoptries: an arsenal of secreted virulence factors. Current Opinion in Microbiology 10, 582–587.CrossRefGoogle ScholarPubMed
Canepa, GE, Molina-Cruz, A, Yenkoidiok-Douti, L, Calvo, E, Williams, AE, Burkhardt, M, Peng, F, Narum, D, Boulanger, MJ, Valenzuela, JG and Barillas-Mury, C (2018) Antibody targeting of a specific region of Pfs47 blocks Plasmodium falciparum malaria transmission. NPJ Vaccines 3, 26.CrossRefGoogle ScholarPubMed
Carias, LL, Dechavanne, S, Nicolete, VC, Sreng, S, Suon, S, Amaratunga, C, Fairhurst, RM, Dechavanne, C, Barnes, S, Witkowski, B, Popovici, J, Roesch, C, Chen, E, Ferreira, MU, Tolia, NH, Adams, JH and King, CL (2019) Identification and characterization of functional human monoclonal antibodies to plasmodium vivax Duffy-binding protein. Journal of Immunology 202, 2648–2660.CrossRefGoogle ScholarPubMed
Chaichan, P, Mercier, A, Galal, L, Mahittikorn, A, Ariey, F, Morand, S, Boumediene, F, Udonsom, R, Hamidovic, A, Murat, JB, Sukthana, Y and Darde, ML (2017) Geographical distribution of Toxoplasma gondii genotypes in Asia: a link with neighboring continents. Infection Genetics and Evolution 53, 227–238.CrossRefGoogle ScholarPubMed
Chen, CW and Chang, CY (2017) Peptide scanning-assisted identification of a monoclonal antibody-recognized linear B-cell epitope. Journal of Visualized Experiments: JoVE 121, e55417.Google Scholar
Chen, ZW, Gao, JM, Huo, XX, Wang, L, Yu, L, Halm-Lai, F, Xu, YH, Song, WJ, Hide, G, Shen, JL and Lun, ZR (2011) Genotyping of Toxoplasma gondii isolates from cats in different geographic regions of China. Veterinary Parasitology 183, 166–170.CrossRefGoogle ScholarPubMed
Chiou, SP, Kitoh, K, Igarashi, I and Takashima, Y (2014) Generation of monoclonal autoantibodies from Babesia rodhaini-infected mice. Journal of Veterinary Medical Science 76, 1281–1284.CrossRefGoogle ScholarPubMed
Cunha-Junior, JP, Silva, DA, Silva, NM, Souza, MA, Souza, GR, Prudencio, CR, Pirovani, CP, Cezar, M, Cascardo, J, Barbosa, BF, Goulart, LR and Mineo, JR (2010) A4d12 monoclonal antibody recognizes a new linear epitope from SAG2A Toxoplasma gondii tachyzoites, identified by phage display bioselection. Immunobiology 215, 26–37.CrossRefGoogle ScholarPubMed
Dubey, JP (2008) The history of Toxoplasma gondii – the first 100 years. Journal of Eukaryotic Microbiology 55, 467–475.CrossRefGoogle ScholarPubMed
Elsheikha, HM (2008) Congenital toxoplasmosis: priorities for further health promotion action. Public Health 122, 335–353.CrossRefGoogle ScholarPubMed
Fentress, SJ, Behnke, MS, Dunay, IR, Mashayekhi, M, Rommereim, LM, Fox, BA, Bzik, DJ, Taylor, GA, Turk, BE, Lichti, CF, Townsend, RR, Qiu, W, Hui, R, Beatty, WL and Sibley, LD (2010) Phosphorylation of immunity-related GTPases by a Toxoplasma gondii-secreted kinase promotes macrophage survival and virulence. Cell Host & Microbe 8, 484–495.CrossRefGoogle ScholarPubMed
Gao, Y, Huang, X, Zhu, Y and Lv, Z (2018) A brief review of monoclonal antibody technology and its representative applications in immunoassays. Journal of Immunoassay & Immunochemistry 39, 351–364.CrossRefGoogle ScholarPubMed
Gondim, LF, Wolf, A, Vrhovec, MG, Pantchev, N, Bauer, C, Langenmayer, MC, Bohne, W, Teifke, JP, Dubey, JP, Conraths, FJ and Schares, G (2016) Characterization of an IgG monoclonal antibody targeted to both tissue cyst and sporocyst walls of Toxoplasma gondii. Experimental Parasitology 163, 46–56.CrossRefGoogle Scholar
Hakimi, H, Goto, Y, Suganuma, K, Angeles, JM, Kawai, S, Inoue, N and Kawazu, S (2015) Development of monoclonal antibodies against Plasmodium falciparum thioredoxin peroxidase 1 and its possible application for malaria diagnosis. Experimental Parasitology 154, 62–66.CrossRefGoogle ScholarPubMed
Hill, D and Dubey, JP (2002) Toxoplasma gondii: transmission, diagnosis and prevention. Clinical Microbiology and Infection 8, 634–640.CrossRefGoogle ScholarPubMed
Jensen, KD, Camejo, A, Melo, MB, Cordeiro, C, Julien, L, Grotenbreg, GM, Frickel, EM, Ploegh, HL, Young, L and Saeij, JP (2015) Toxoplasma gondii superinfection and virulence during secondary infection correlate with the exact ROP5/ROP18 allelic combination. MBio 6, e02280.CrossRefGoogle ScholarPubMed
Jones, JL and Dubey, JP (2012) Foodborne toxoplasmosis. Clinical Infectious Diseases 55, 845–851.CrossRefGoogle ScholarPubMed
Jones, M, McLoughlin, V, Connolly, JG, Farquhar, CF, MacGregor, IR and Head, MW (2009) Production and characterization of a panel of monoclonal antibodies against native human cellular prion protein. Hybridoma (Larchmt) 28, 13–20.CrossRefGoogle ScholarPubMed
Kohler, G and Milstein, C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256, 495–497.CrossRefGoogle ScholarPubMed
Kong, JT, Grigg, ME, Uyetake, L, Parmley, S and Boothroyd, JC (2003) Serotyping of Toxoplasma gondii infections in humans using synthetic peptides. Journal of Infectious Diseases 187, 1484–1495.CrossRefGoogle ScholarPubMed
Li, YS, Ross, AG, Sleigh, AC, Li, Y, Waine, GJ, Williams, GJ, Tanner, M and McManus, DP (1999) Antibody isotype responses, infection and re-infection for Schistosoma japonicum in a marshland area of China. Acta Tropica 73, 79–92.CrossRefGoogle Scholar
Liu, Q, Chen, Z, Shi, W, Sun, H, Zhang, J, Li, H, Xiao, Y, Wang, F and Zhao, X (2014) Preparation and initial application of monoclonal antibodies that recognize Eimeria tenella microneme proteins 1 and 2. Parasitology Research 113, 4151–4161.CrossRefGoogle ScholarPubMed
Liu, GY, Mei, XJ, Hu, MJ, Yang, Y, Liu, M, Li, MS, Zhang, ML, Cao, MJ and Liu, GM (2018) Analysis of the allergenic epitopes of tropomyosin from mud crab using phage display and site-directed mutagenesis. Journal of Agricultural and Food Chemistry 66, 9127–9137.CrossRefGoogle ScholarPubMed
Mei, S, Li, F, Leier, A, Marquez-Lago, TT, Giam, K, Croft, NP, Akutsu, T, Smith, AI, Li, J, Rossjohn, J, Purcell, AW and Song, J (2019) A comprehensive review and performance evaluation of bioinformatics tools for HLA class I peptide-binding prediction. Briefings in Bioinformatics 00, 1–17.Google Scholar
Montoya, JG and Remington, JS (2008) Management of Toxoplasma gondii infection during pregnancy. Clinical Infectious Diseases 47, 554–566.CrossRefGoogle ScholarPubMed
Parmley, SF, Gross, U, Sucharczuk, A, Windeck, T, Sgarlato, GD and Remington, JS (1994) Two alleles of the gene encoding surface antigen P22 in 25 strains of Toxoplasma gondii. Journal of Parasitology 80, 293–301.CrossRefGoogle ScholarPubMed
Purcell, AW and Gorman, JJ (2004) Immunoproteomics: mass spectrometry-based methods to study the targets of the immune response. Molecular and Cellular Proteomics 3, 193–208.CrossRefGoogle Scholar
Ratcliffe, EC and Wilson, RA (1991) The magnitude and kinetics of delayed-type hypersensitivity responses in mice vaccinated with irradiated cercariae of Schistosoma mansoni. Parasitology 103(Pt 1), 65–75.CrossRefGoogle ScholarPubMed
Robbins, JR, Zeldovich, VB, Poukchanski, A, Boothroyd, JC and Bakardjiev, AI (2012) Tissue barriers of the human placenta to infection with Toxoplasma gondii. Infection and Immunity 80, 418–428.CrossRefGoogle ScholarPubMed
Saeij, JP, Boyle, JP, Coller, S, Taylor, S, Sibley, LD, Brooke-Powell, ET, Ajioka, JW and Boothroyd, JC (2006) Polymorphic secreted kinases are key virulence factors in toxoplasmosis. Science 314, 1780–1783.CrossRefGoogle ScholarPubMed
Sibley, LD, Qiu, W, Fentress, S, Taylor, SJ, Khan, A and Hui, R (2009) Forward genetics in Toxoplasma gondii reveals a family of rhoptry kinases that mediates pathogenesis. Eukaryotic Cell 8, 1085–1093.CrossRefGoogle ScholarPubMed
Sinai, AP (2007) The toxoplasma kinase ROP18: an active member of a degenerate family. PLoS Pathogens 3, e16.CrossRefGoogle ScholarPubMed
Steinfeldt, T, Konen-Waisman, S, Tong, L, Pawlowski, N, Lamkemeyer, T, Sibley, LD, Hunn, JP and Howard, JC (2010) Phosphorylation of mouse immunity-related GTPase (IRG) resistance proteins is an evasion strategy for virulent Toxoplasma gondii. PLoS Biology 8, e1000576.CrossRefGoogle ScholarPubMed
Taylor, S, Barragan, A, Su, C, Fux, B, Fentress, SJ, Tang, K, Beatty, WL, Hajj, HE, Jerome, M, Behnke, MS, White, M, Wootton, JC and Sibley, LD (2006) A secreted serine-threonine kinase determines virulence in the eukaryotic pathogen Toxoplasma gondii. Science 314, 1776–1780.CrossRefGoogle ScholarPubMed
Trager, W and Jensen, JB (1976) Human malaria parasites in continuous culture. Science 193, 673–675.CrossRefGoogle ScholarPubMed
Virreira Winter, S, Niedelman, W, Jensen, KD, Rosowski, EE, Julien, L, Spooner, E, Caradonna, K, Burleigh, BA, Saeij, JP, Ploegh, HL and Frickel, EM (2011) Determinants of GBP recruitment to Toxoplasma gondii vacuoles and the parasitic factors that control it. PLoS ONE 6, e24434.CrossRefGoogle ScholarPubMed
Xia, J, Kong, L, Zhou, LJ, Wu, SZ, Yao, LJ, He, C, He, CY and Peng, HJ (2018) Genome-wide bimolecular fluorescence complementation-based proteomic analysis of Toxoplasma gondii ROP18's human interactome shows its key role in regulation of cell immunity and apoptosis. Frontiers in Immunology 9, 61.CrossRefGoogle ScholarPubMed
Yamamoto, M and Takeda, K (2012) Inhibition of ATF6beta-dependent host adaptive immune response by a Toxoplasma virulence factor ROP18. Virulence 3, 77–80.CrossRefGoogle ScholarPubMed
Yamamoto, M, Ma, JS, Mueller, C, Kamiyama, N, Saiga, H, Kubo, E, Kimura, T, Okamoto, T, Okuyama, M, Kayama, H, Nagamune, K, Takashima, S, Matsuura, Y, Soldati-Favre, D and Takeda, K (2011) ATF6beta Is a host cellular target of the Toxoplasma gondii virulence factor ROP18. Journal of Experimental Medicine 208, 1533–1546.CrossRefGoogle ScholarPubMed
Yang, Z, Hou, Y, Hao, T, Rho, HS, Wan, J, Luan, Y, Gao, X, Yao, J, Pan, A, Xie, Z, Qian, J, Liao, W, Zhu, H and Zhou, X (2017) A human proteome array approach to identifying key host proteins targeted by toxoplasma kinase ROP18. Molecular & Cellular Proteomics 16, 469–484.CrossRefGoogle ScholarPubMed
Yin, K, Zhao, G, Xu, C, Qiu, X, Wen, B, Sun, H, Liu, G, Liu, Y, Zhao, Q, Wei, Q, Huang, B, Yan, G and Cao, J (2019) Prediction of Toxoplasma gondii virulence factor ROP18 competitive inhibitors by virtual screening. Parasites & Vectors 12, 98.CrossRefGoogle ScholarPubMed
Yuan, ZG, Zhang, XX, Lin, RQ, Petersen, E, He, S, Yu, M, He, XH, Zhou, DH, He, Y, Li, HX, Liao, M and Zhu, XQ (2011) Protective effect against toxoplasmosis in mice induced by DNA immunization with gene encoding Toxoplasma gondii ROP18. Vaccine 29, 6614–6619.CrossRefGoogle ScholarPubMed
Zhu, W, Li, J, Pappoe, F, Shen, J and Yu, L (2019) Strategies developed by Toxoplasma gondii to survive in the host. Frontiers in Microbiology 10, 899.CrossRefGoogle ScholarPubMed
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