Epitope Capsid-Incorporation: New Effective Approach for Vaccine Development for Chagas Disease

Qiana L. Matthews, Anitra L. Farrow, Girish Rachakonda, Linlin Gu, Pius Nde, Alexandre Krendelchtchikov, Siddarth Pratap, Shruti S. Sakhare, Steffanie Sabbaj, Maria F. Lima, Fernando Villalta

Abstract


Background: Previously we reported that a hexon-modified adenovirus (Ad) vector containing the invasive neutralizing epitope of Trypanosoma cruzi (T. cruzi) trypomastigote gp83 (Ad5-gp83) provided immunoprotection against T. cruzi infection. The purpose of this work was to design an improved vaccine for T. cruzi using a novel epitope capsid incorporation strategy. Thus, we evaluated the immunoprotection raised by co-immunization with Ad5-gp83 and an Ad vector containing an epitope (ASP-M) of the T. cruzi amastigote surface protein 2.

Methods: Protein IX (pIX)-modified Ad vector (Ad5-pIX-ASP-M) was generated, characterized, and validated. C3H/He mice were immunized with Ad5-pIX-ASP-M and Ad5-gp83 and the cell-mediated responses were evaluated by enzyme-linked immunospot (ELISPOT) assay and intracellular staining. Immunized mice were challenged with T. cruzi to evaluate the vaccine efficacy.


Results: Our findings indicate that Ad5-pIX-ASP-M was viable. Specific CD8+ T-cell mediated responses prior to the challenge show an increase in IFNγ and TNFα production. A single immunization with Ad5-pIX-ASP-M provided protection from T. cruzi infection, but co-immunizations with Ad5-pIX-ASP-M and Ad5-gp83 provided a higher immunoprotection and increased survival rate of mice.

Conclusions: Overall, these results suggest that the combination of gp83 and ASP-M specific epitopes onto the capsid-incorporated adenoviruses would provide superior protection against Chagas disease as compared with Ad5-gp83 alone.


Keywords


Chagas disease; T. cruzi vaccine constructs; epitope-capsid incorporation; trypomastigote gp83 neutralizing epitope; amastigote surface protein 2 epitope; co-immunization; immunoprotection

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References


1. Coura JR, Vinas PA. Chagas disease: a new worldwide challenge. Nature. 2010;465(7301):S6-7. PubMed PMID: 20571554. doi: 10.1038/nature09221

2. Tanowitz HB, Weiss LM, Montgomery SP. Chagas disease has now gone global. PLoS Negl Trop Dis. 2011;5(4):e1136. PubMed PMID: 21572510. Pubmed Central PMCID: 3082509. 10.1371/journal.pntd.0001136

3. Hotez PJ, Dumonteil E, Betancourt Cravioto M, Bottazzi ME, Tapia-Conyer R, Meymandi S, Karunakara U, Ribeiro I, Cohen RM, Pecoul B. An unfolding tragedy of Chagas disease in North America. PLoS Negl Trop Dis. 2013;7(10):e2300. PubMed PMID: 24205411. Pubmed Central PMCID: 3814410. 10.1371/journal.pntd.0002300

4. Lee BY, Bacon KM, Bottazzi ME, Hotez PJ. Global economic burden of Chagas disease: a computational simulation model. Lancet Infect Dis. 2013;13(4):342-8. PubMed PMID: 23395248. Pubmed Central PMCID: 3763184. doi: 10.1016/S1473-3099(13)70002-1

5. Chatelain E. Chagas disease drug discovery: toward a new era. J Biomol Screen. 2015;20(1):22-35. PubMed PMID: 25245987. doi: 10.1177/1087057114550585

6. Molina I, Gomez i Prat J, Salvador F, Trevino B, Sulleiro E, Serre N, Pou D, Roure S, Cabezos J, Valerio L, Blanco-Grau A, Sanchez-Montalva A, Vidal X, Pahissa A. Randomized trial of posaconazole and benznidazole for chronic Chagas' disease. N Engl J Med. 2014;370(20):1899-908. PubMed PMID: 24827034. doi: 10.1056/NEJMoa1313122

7. Villalta F, Dobish MC, Nde PN, Kleshchenko YY, Hargrove TY, Johnson CA, Waterman MR, Johnston JN, Lepesheva GI. VNI cures acute and chronic experimental Chagas disease. J Infect Dis. 2013;208(3):504-11. PubMed PMID: 23372180. Pubmed Central PMCID: 3698996. doi: 10.1093/infdis/jit042

8. Lepesheva GI, Hargrove TY, Rachakonda G, Wawrzak Z, Pomel S, Cojean S, Nde PN, Nes WD, Locuson CW, Calcutt MW, Waterman MR, Daniels JS, Loiseau PM, Villalta F. VFV as a New Effective CYP51 Structure-Derived Drug Candidate for Chagas Disease and Visceral Leishmaniasis. J Infect Dis. 2015;212(9):1439-48. PubMed PMID: 25883390. Pubmed Central PMCID: PMC4601915. doi: 10.1093/infdis/jiv228

9. Teixeira AR, Hecht MM, Guimaro MC, Sousa AO, Nitz N. Pathogenesis of chagas' disease: parasite persistence and autoimmunity. Clin Microbiol Rev. 2011;24(3):592-630. PubMed PMID: 21734249. Pubmed Central PMCID: 3131057. doi: 10.1128/CMR.00063-10

10. Rassi A, Jr., Rassi A, Marin-Neto JA. Chagas disease. Lancet. 2010;375(9723):1388-402. PubMed PMID: 20399979. doi: 10.1016/S0140-6736(10)60061-X

11. Marin-Neto JA, Cunha-Neto E, Maciel BC, Simoes MV. Pathogenesis of chronic Chagas heart disease. Circulation. 2007;115(9):1109-23. PubMed PMID: 17339569. doi:10.1161/CIRCULATIONAHA.106.624296

12. Gutierrez FR, Guedes PM, Gazzinelli RT, Silva JS. The role of parasite persistence in pathogenesis of Chagas heart disease. Parasite Immunol. 2009;31(11):673-85. PubMed PMID: 19825107. doi: 10.1111/j.1365-3024.2009.01108.x

13. Girones N, Cuervo H, Fresno M. Trypanosoma cruzi-induced molecular mimicry and Chagas' disease. Curr Top Microbiol Immunol. 2005;296:89-123. PubMed PMID: 16323421.

14. Iwai LK, Juliano MA, Juliano L, Kalil J, Cunha-Neto E. T-cell molecular mimicry in Chagas disease: identification and partial structural analysis of multiple cross-reactive epitopes between Trypanosoma cruzi B13 and cardiac myosin heavy chain. J Autoimmun. 2005;24(2):111-7. PubMed PMID: 15829403. doi: 10.1016/j.jaut.2005.01.006

15. Cunha-Neto E, Bilate AM, Hyland KV, Fonseca SG, Kalil J, Engman DM. Induction of cardiac autoimmunity in Chagas heart disease: a case for molecular mimicry. Autoimmunity. 2006;39(1):41-54. PubMed PMID: 16455581. doi: 10.1080/08916930500485002

16. Lopes MF, DosReis GA. Trypanosoma cruzi-induced immunosuppression: selective triggering of CD4+ T-cell death by the T-cell receptor-CD3 pathway and not by the CD69 or Ly-6 activation pathway. Infect Immun. 1996;64(5):1559-64. PubMed PMID: 8613360. Pubmed Central PMCID: PMC173961.

17. Ouaissi A, Da Silva AC, Guevara AG, Borges M, Guilvard E. Trypanosoma cruzi-Induced Host Immune System Dysfunction: A Rationale for Parasite Immunosuppressive Factor(s) Encoding Gene Targeting. J Biomed Biotechnol. 2001;1(1):11-7. PubMed PMID: 12488621. Pubmed Central PMCID: PMC79673. doi: 10.1155/S1110724301000055

18. Villalta F, Kierszenbaum F. Immunization against a challenge with insect vector, metacyclic forms of Trypanosoma cruzi simulating a natural infection. Am J Trop Med Hyg. 1983;32(2):273-6. PubMed PMID: 6404187.

19. Cazorla SI, Frank FM, Malchiodi EL. Vaccination approaches against Trypanosoma cruzi infection. Expert Rev Vaccines. 2009;8(7):921-35. PubMed PMID: 19538117. doi:10.1586/erv.09.45

20. Dumonteil E, Bottazzi ME, Zhan B, Heffernan MJ, Jones K, Valenzuela JG, Kamhawi S, Ortega J, Rosales SP, Lee BY, Bacon KM, Fleischer B, Slingsby BT, Cravioto MB, Tapia-Conyer R, Hotez PJ. Accelerating the development of a therapeutic vaccine for human Chagas disease: rationale and prospects. Expert Rev Vaccines. 2012;11(9):1043-55. PubMed PMID: 23151163. Pubmed Central PMCID: PMC3819810. doi: 10.1586/erv.12.85

21. Quijano-Hernandez I, Dumonteil E. Advances and challenges towards a vaccine against Chagas disease. Hum Vaccin. 2011;7(11):1184-91. PubMed PMID: 22048121. Pubmed Central PMCID: PMC3323496. doi: 10.4161/hv.7.11.17016

22. Farrow AL, Rachakonda G, Gu L, Krendelchtchikova V, Nde PN, Pratap S, Lima MF, Villalta F, Matthews QL. Immunization with Hexon modified adenoviral vectors integrated with gp83 epitope provides protection against Trypanosoma cruzi infection. PLoS Negl Trop Dis. 2014;8(8):e3089. PubMed PMID: 25144771. Pubmed Central PMCID: 4140675. doi: 10.1371/journal.pntd.0003089

23. Villalta F, Smith CM, Ruiz-Ruano A, Lima MF. A ligand that Trypanosoma cruzi uses to bind to mammalian cells to initiate infection. FEBS Lett. 2001;505(3):383-8. PubMed PMID: 11576533.

24. Villalta F, Lima MF, Ruiz-Ruano A, Zhou L. Attachment of Trypanosoma cruzi to host cells: a monoclonal antibody recognizes a trypomastigote stage-specific epitope on the gp 83 required for parasite attachment. Biochem Biophys Res Commun. 1992;182(1):6-13. PubMed PMID: 1370617.

25. Pan AA, McMahon-Pratt D. Amastigote and epimastigote stage-specific components of Trypanosoma cruzi characterized by using monoclonal antibodies. Purification and molecular characterization of an 83-kilodalton amastigote protein. J Immunol. 1989;143(3):1001-8. PubMed PMID: 2501384.

26. Low HP, Tarleton RL. Molecular cloning of the gene encoding the 83 kDa amastigote surface protein and its identification as a member of the Trypanosoma cruzi sialidase superfamily. Mol Biochem Parasitol. 1997;88(1-2):137-49. PubMed PMID: 9274875.

27. Low HP, Santos MA, Wizel B, Tarleton RL. Amastigote surface proteins of Trypanosoma cruzi are targets for CD8+ CTL. J Immunol. 1998;160(4):1817-23. PubMed PMID: 9469442.

28. Machado AV, Cardoso JE, Claser C, Rodrigues MM, Gazzinelli RT, Bruna-Romero O. Long-term protective immunity induced against Trypanosoma cruzi infection after vaccination with recombinant adenoviruses encoding amastigote surface protein-2 and trans-sialidase. Hum Gene Ther. 2006;17(9):898-908. PubMed PMID: 16972758. doi:10.1089/hum.2006.17.898

29. Barbosa RP, Filho BG, Dos Santos LI, Junior PA, Marques PE, Pereira RV, Cara DC, Bruna-Romero O, Rodrigues MM, Gazzinelli RT, Machado AV. Vaccination using recombinants influenza and adenoviruses encoding amastigote surface protein-2 are highly effective on protection against Trypanosoma cruzi infection. PLoS One. 2013;8(4):e61795. PubMed PMID: 23637908. Pubmed Central PMCID: 3634828. doi: 10.1371/journal.pone.0061795

30. Lima MF, Villalta F. Trypanosoma cruzi trypomastigote clones differentially express a parasite cell adhesion molecule. Mol Biochem Parasitol. 1989;33(2):159-70. PubMed PMID: 2657421.

31. Lepesheva GI, Hargrove TY, Anderson S, Kleshchenko Y, Furtak V, Wawrzak Z, Villalta F, Waterman MR. Structural insights into inhibition of sterol 14alpha-demethylase in the human pathogen Trypanosoma cruzi. J Biol Chem. 2010;285(33):25582-90. PubMed PMID: 20530488. Pubmed Central PMCID: 2919122. doi: 10.1074/jbc.M110.133215

32. Matthews QL, Sibley DA, Wu H, Li J, Stoff-Khalili MA, Waehler R, Mathis JM, Curiel DT. Genetic incorporation of a herpes simplex virus type 1 thymidine kinase and firefly luciferase fusion into the adenovirus protein IX for functional display on the virion. Mol Imaging. 2006;5(4):510-9. PubMed PMID: 17150163. Pubmed Central PMCID: 1781529.

33. Gu L, Li ZC, Krendelchtchikov A, Krendelchtchikova V, Wu H, Matthews QL. Using multivalent adenoviral vectors for HIV vaccination. PLoS One. 2013;8(3):e60347. PubMed PMID: 23555957. Pubmed Central PMCID: 3610663. doi: 10.1371/journal.pone.0060347

34. Wu H, Han T, Belousova N, Krasnykh V, Kashentseva E, Dmitriev I, Kataram M, Mahasreshti PJ, Curiel DT. Identification of sites in adenovirus hexon for foreign peptide incorporation. J Virol. 2005;79(6):3382-90. PubMed PMID: 15731232. Pubmed Central PMCID: 1075677. doi: 10.1128/JVI.79.6.3382-3390.2005

35. Johnson CA, Rachakonda G, Kleshchenko YY, Nde PN, Madison MN, Pratap S, Cardenas TC, Taylor C, Lima MF, Villalta F. Cellular response to Trypanosoma cruzi infection induces secretion of defensin alpha-1, which damages the flagellum, neutralizes trypanosome motility, and inhibits infection. Infect Immun. 2013;81(11):4139-48. PubMed PMID: 23980110. Pubmed Central PMCID: 3811821. doi: 10.1128/IAI.01459-12

36. Araujo AF, de Alencar BC, Vasconcelos JR, Hiyane MI, Marinho CR, Penido ML, Boscardin SB, Hoft DF, Gazzinelli RT, Rodrigues MM. CD8+-T-cell-dependent control of Trypanosoma cruzi infection in a highly susceptible mouse strain after immunization with recombinant proteins based on amastigote surface protein 2. Infect Immun. 2005;73(9):6017-25. PubMed PMID: 16113322. Pubmed Central PMCID: PMC1231112. doi: 10.1128/IAI.73.9.6017-6025.2005

37. Sargent KL, Meulenbroek RA, Parks RJ. Activation of adenoviral gene expression by protein IX is not required for efficient virus replication. J Virol. 2004;78(10):5032-7. PubMed PMID: 15113884. Pubmed Central PMCID: PMC400331.

38. Guidotti LG, Ishikawa T, Hobbs MV, Matzke B, Schreiber R, Chisari FV. Intracellular inactivation of the hepatitis B virus by cytotoxic T lymphocytes. Immunity. 1996;4(1):25-36. PubMed PMID: 8574849.

39. Chyuan IT, Tsai HF, Tzeng HT, Sung CC, Wu CS, Chen PJ, Hsu PN. Tumor necrosis factor-alpha blockage therapy impairs hepatitis B viral clearance and enhances T-cell exhaustion in a mouse model. Cell Mol Immunol. 2015;12(3):317-25. PubMed PMID: 25661729. doi: 10.1038/cmi.2015.01

40. Kasahara S, Ando K, Saito K, Sekikawa K, Ito H, Ishikawa T, Ohnishi H, Seishima M, Kakumu S, Moriwaki H. Lack of tumor necrosis factor alpha induces impaired proliferation of hepatitis B virus-specific cytotoxic T lymphocytes. J Virol. 2003;77(4):2469-76. PubMed PMID: 12551985. Pubmed Central PMCID: 141095.

41. Thacker EE, Timares L, Matthews QL. Strategies to overcome host immunity to adenovirus vectors in vaccine development. Expert Rev Vaccines. 2009;8(6):761-77. PubMed PMID: 19485756. Pubmed Central PMCID: 3700409. doi: 10.1586/erv.09.29

42. Chirmule N, Propert K, Magosin S, Qian Y, Qian R, Wilson J. Immune responses to adenovirus and adeno-associated virus in humans. Gene Ther. 1999;6(9):1574-83. PubMed PMID: 10490767. doi: 10.1038/sj.gt.3300994

43. Schagen FH, Ossevoort M, Toes RE, Hoeben RC. Immune responses against adenoviral vectors and their transgene products: a review of strategies for evasion. Crit Rev Oncol Hematol. 2004;50(1):51-70. PubMed PMID: 15094159. doi: 10.1016/S1040-8428(03)00172-0

44. Zaiss AK, Machado HB, Herschman HR. The influence of innate and pre-existing immunity on adenovirus therapy. J Cell Biochem. 2009;108(4):778-90. PubMed PMID: 19711370. Pubmed Central PMCID: 2822460. doi: 10.1002/jcb.22328

45. Tian X, Su X, Li H, Li X, Zhou Z, Liu W, Zhou R. Construction and characterization of human adenovirus serotype 3 packaged by serotype 7 hexon. Virus Res. 2011;160(1-2):214-20. PubMed PMID: 21740937. doi: 10.1016/j.virusres.2011.06.017

46. Soloff AC, Liu X, Gao W, Day RD, Gambotto A, Barratt-Boyes SM. Adenovirus 5- and 35-based immunotherapy enhances the strength but not breadth or quality of immunity during chronic SIV infection. Eur J Immunol. 2009;39(9):2437-49. PubMed PMID: 19670380. doi: 10.1002/eji.200839130

47. Abbink P, Lemckert AA, Ewald BA, Lynch DM, Denholtz M, Smits S, Holterman L, Damen I, Vogels R, Thorner AR, O'Brien KL, Carville A, Mansfield KG, Goudsmit J, Havenga MJ, Barouch DH. Comparative seroprevalence and immunogenicity of six rare serotype recombinant adenovirus vaccine vectors from subgroups B and D. J Virol. 2007;81(9):4654-63. PubMed PMID: 17329340. Pubmed Central PMCID: 1900173. doi: 10.1128/JVI.02696-06


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Copyright (c) 2016 Qiana L. Matthews, Anitra L. Farrow, Girish Rachakonda, Linlin Gu, Pius Nde, Alexandre Krendelchtchikov, Siddharth Pratap, Shruti S. Sakhare, Steffanie Sabbaj, Maria F. Lima, Fernando Villalta

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