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Intranasal Vaccination with Outer-Membrane Protein of Orientia tsutsugamushi induces Protective Immunity Against Scrub Typhus

  • Sung-Moo Park (Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University) ;
  • Min Jeong Gu (Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University) ;
  • Young-Jun Ju (Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University) ;
  • In Su Cheon (Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University) ;
  • Kyu-Jam Hwang (Division of Zoonotic and Vector Borne Disease Research, Center for Infectious Disease Research, National Institute of Health) ;
  • Byoungchul Gill (Division of Zoonotic and Vector Borne Disease Research, Center for Infectious Disease Research, National Institute of Health) ;
  • Byoung-Shik Shim (Laboratory Sciences Division, International Vaccine Institute) ;
  • Hang-Jin Jeong (Division of Zoonotic and Vector Borne Disease Research, Center for Infectious Disease Research, National Institute of Health) ;
  • Young Min Son (Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University) ;
  • Sangho Choi (Division of Zoonotic and Vector Borne Disease Research, Center for Infectious Disease Research, National Institute of Health) ;
  • Woonhee Jeung (Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University) ;
  • Seung Hyun Han (Department of Oral Microbiology and Immunology, DRI, and BK21 Plus Program, School of Dentistry, Seoul National University) ;
  • Hyuk Chu (Division of Zoonotic and Vector Borne Disease Research, Center for Infectious Disease Research, National Institute of Health) ;
  • Cheol-Heui Yun (Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University)
  • Received : 2020.03.16
  • Accepted : 2020.11.02
  • Published : 2021.04.30
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Abstract

Scrub typhus develops after the individual is bitten by a trombiculid mite infected with Orientia tsutsugamushi. Since it has been reported that pneumonia is frequently observed in patients with scrub typhus, we investigated whether intranasal (i.n.) vaccination with the outer membrane protein of O. tsutsugamushi (OMPOT) would induce a protective immunity against O. tsutsugamushi infection. It was particular interest that when mice were infected with O. tsutsugamushi, the bacteria disseminated into the lungs, causing pneumonia. The i.n. vaccination with OMPOT induced IgG responses in serum and bronchoalveolar lavage (BAL) fluid. The anti-O. tsutsugamushi IgA Abs in BAL fluid after the vaccination showed a high correlation of the protection against O. tsutsugamushi. The vaccination induced strong Ag-specific Th1 and Th17 responses in the both spleen and lungs. In conclusion, the current study demonstrated that i.n. vaccination with OMPOT elicited protective immunity against scrub typhus in mouse with O. tsutsugamushi infection causing subsequent pneumonia.

Keywords

Acknowledgement

This work was carried out with the support of Cooperative Research Program for Agriculture Science and Technology Development (PJ01336401), Rural Development Administration, and National Research Foundation of Korea (2018R1A2B2006793), Republic of Korea. This research was also supported by a fund (2010-NI52001-00) from Research of Korea Centers for Disease Control and Prevention, and Korea Institute of Planning and Evaluation for Technology in Food, Agriculture and Forestry (IPET) through Animal Disease Management Technology Development Program, funded by Ministry of Agriculture, Food and Rural Affairs (MAFRA) (319081-03). The current work was also partially supported by the BK21 FOUR Program of the Department of Agricultural Biotechnology, Seoul National University, Seoul, Korea.

References

  1. Seong SY, Choi MS, Kim IS. Orientia tsutsugamushi infection: overview and immune responses. Microbes Infect 2001;3:11-21.
  2. Chattopadhyay S, Richards AL. Scrub typhus vaccines: past history and recent developments. Hum Vaccin 2007;3:73-80. https://doi.org/10.4161/hv.3.3.4009
  3. Watt G, Parola P. Scrub typhus and tropical rickettsioses. Curr Opin Infect Dis 2003;16:429-436. https://doi.org/10.1097/00001432-200310000-00009
  4. Kelly DJ, Fuerst PA, Ching WM, Richards AL. Scrub typhus: the geographic distribution of phenotypic and genotypic variants of Orientia tsutsugamushi. Clin Infect Dis 2009;48 Suppl 3:S203-S230. https://doi.org/10.1086/596576
  5. Watt G, Chouriyagune C, Ruangweerayud R, Watcharapichat P, Phulsuksombati D, Jongsakul K, Teja-Isavadharm P, Bhodhidatta D, Corcoran KD, Dasch GA, et al. Scrub typhus infections poorly responsive to antibiotics in northern Thailand. Lancet 1996;348:86-89. https://doi.org/10.1016/S0140-6736(96)02501-9
  6. Asadi Karam MR, Habibi M, Bouzari S. Use of flagellin and cholera toxin as adjuvants in intranasal vaccination of mice to enhance protective immune responses against uropathogenic Escherichia coli antigens. Biologicals 2016;44:378-386. https://doi.org/10.1016/j.biologicals.2016.06.006
  7. Fulton F, Joyner L. Cultivation of Rickettsia tsutsugamushi in lungs of rodents; preparation of a scrub-typhus vaccine. Lancet 1945;2:729-734. https://doi.org/10.1016/S0140-6736(45)91069-5
  8. Chattopadhyay S, Jiang J, Chan TC, Manetz TS, Chao CC, Ching WM, Richards AL. Scrub typhus vaccine candidate Kp r56 induces humoral and cellular immune responses in cynomolgus monkeys. Infect Immun 2005;73:5039-5047. https://doi.org/10.1128/IAI.73.8.5039-5047.2005
  9. Varghese GM, Trowbridge P, Janardhanan J, Thomas K, Peter JV, Mathews P, Abraham OC, Kavitha ML. Clinical profile and improving mortality trend of scrub typhus in South India. Int J Infect Dis 2014;23:39-43. https://doi.org/10.1016/j.ijid.2014.02.009
  10. Hsu YH, Chen HI. Pulmonary pathology in patients associated with scrub typhus. Pathology 2008;40:268-271. https://doi.org/10.1080/00313020801911488
  11. Moron CG, Popov VL, Feng HM, Wear D, Walker DH. Identification of the target cells of Orientia tsutsugamushi in human cases of scrub typhus. Mod Pathol 2001;14:752-759. https://doi.org/10.1038/modpathol.3880385
  12. Urakami H, Tsuruhara T, Tamura A. Penetration of Rickettsia tsutsugamushi into cultured mouse fibroblasts (L cells): an electron microscopic observation. Microbiol Immunol 1983;27:251-263. https://doi.org/10.1111/j.1348-0421.1983.tb03587.x
  13. Song SW, Kim KT, Ku YM, Park SH, Kim YS, Lee DG, Yoon SA, Kim YO. Clinical role of interstitial pneumonia in patients with scrub typhus: a possible marker of disease severity. J Korean Med Sci 2004;19:668-673. https://doi.org/10.3346/jkms.2004.19.5.668
  14. Chan CH, Lai F, Daly B, Ho AK, Yu CM, Cheng A. Scrub typhus pneumonitis with delayed resolution. J Trop Med Hyg 1995;98:114-116.
  15. Chayakul P, Panich V, Silpapojakul K. Scrub typhus pneumonitis: an entity which is frequently missed. Q J Med 1988;68:595-602.
  16. Smadel JE, Ley HL Jr, Diercks FH, Paterson PY, Wisseman CL Jr, Traub R. Immunization against scrub typhus: duration of immunity in volunteers following combined living vaccine and chemoprophylaxis. Am J Trop Med Hyg 1952;1:87-99. https://doi.org/10.4269/ajtmh.1952.1.87
  17. Ni YS, Chan TC, Chao CC, Richards AL, Dasch GA, Ching WM. Protection against scrub typhus by a plasmid vaccine encoding the 56-KD outer membrane protein antigen gene. Am J Trop Med Hyg 2005;73:936-941. https://doi.org/10.4269/ajtmh.2005.73.936
  18. Trent B, Liang Y, Xing Y, Esqueda M, Wei Y, Cho NH, Kim HI, Kim YS, Shelite TR, Cai J, et al. Polarized lung inflammation and Tie2/angiopoietin-mediated endothelial dysfunction during severe Orientia tsutsugamushi infection. PLoS Negl Trop Dis 2020;14:e0007675.
  19. Trent B, Fisher J, Soong L. Scrub typhus pathogenesis: innate immune response and lung injury during Orientia tsutsugamushi infection. Front Microbiol 2019;10:2065.
  20. Kim HI, Ha NY, Kim G, Min CK, Kim Y, Yen NTH, Choi MS, Cho NH. Immunization with a recombinant antigen composed of conserved blocks from TSA56 provides broad genotype protection against scrub typhus. Emerg Microbes Infect 2019;8:946-958. https://doi.org/10.1080/22221751.2019.1632676
  21. Ha NY, Kim Y, Min CK, Kim HI, Yen NTH, Choi MS, Kang JS, Kim YS, Cho NH. Longevity of antibody and T-cell responses against outer membrane antigens of Orientia tsutsugamushi in scrub typhus patients. Emerg Microbes Infect 2017;6:e116.
  22. Choi S, Jeong HJ, Hwang KJ, Gill B, Ju YR, Lee YS, Lee J. A recombinant 47-kDa outer membrane protein induces an immune response against Orientia tsutsugamushi strain Boryong. Am J Trop Med Hyg 2017;97:30-37. https://doi.org/10.4269/ajtmh.15-0771
  23. Yu Y, Wen B, Wen B, Niu D, Chen M, Qiu L. Induction of protective immunity against scrub typhus with a 56-kilodalton recombinant antigen fused with a 47-kilodalton antigen of Orientia tsutsugamushi Karp. Am J Trop Med Hyg 2005;72:458-464. https://doi.org/10.4269/ajtmh.2005.72.458
  24. Chen HW, Zhang Z, Huber E, Mutumanje E, Chao CC, Ching WM. Kinetics and magnitude of antibody responses against the conserved 47-kilodalton antigen and the variable 56-kilodalton antigen in scrub typhus patients. Clin Vaccine Immunol 2011;18:1021-1027. https://doi.org/10.1128/CVI.00017-11
  25. Hickman CJ, Stover CK, Joseph SW, Oaks EV. Murine T-cell response to native and recombinant protein antigens of Rickettsia tsutsugamushi. Infect Immun 1993;61:1674-1681. https://doi.org/10.1128/iai.61.5.1674-1681.1993
  26. Oaks EV, Rice RM, Kelly DJ, Stover CK. Antigenic and genetic relatedness of eight Rickettsia tsutsugamushi antigens. Infect Immun 1989;57:3116-3122. https://doi.org/10.1128/iai.57.10.3116-3122.1989
  27. Chen HW, Zhang Z, Huber E, Chao CC, Wang H, Dasch GA, Ching WM. Identification of cross-reactive epitopes on the conserved 47-kilodalton antigen of Orientia tsutsugamushi and human serine protease. Infect Immun 2009;77:2311-2319.
  28. Chu H, Park SM, Cheon IS, Park MY, Shim BS, Gil BC, Jeung WH, Hwang KJ, Song KD, Hong KJ, et al. Orientia tsutsugamushi infection induces CD4+ T cell activation via human dendritic cell activity. J Microbiol Biotechnol 2013;23:1159-1166. https://doi.org/10.4014/jmb.1303.03019
  29. Tamura A, Urakami H. Easy method for infectivity titration of Rickettsia tsutsugamushi by infected cell counting (author's transl). Nippon Saikingaku Zasshi 1981;36:783-785. https://doi.org/10.3412/jsb.36.783
  30. Ha NY, Sharma P, Kim G, Kim Y, Min CK, Choi MS, Kim IS, Cho NH. Immunization with an autotransporter protein of Orientia tsutsugamushi provides protective immunity against scrub typhus. PLoS Negl Trop Dis 2015;9:e0003585.
  31. Min CK, Kim HI, Ha NY, Kim Y, Kwon EK, Yen NTH, Youn JI, Jeon YK, Inn KS, Choi MS, et al. A type I interferon and IL-10 induced by Orientia tsutsugamushi infection suppresses antigen-specific T cells and their memory responses. Front Immunol 2018;9:2022.
  32. Aida Y, Pabst MJ. Removal of endotoxin from protein solutions by phase separation using Triton X-114. J Immunol Methods 1990;132:191-195. https://doi.org/10.1016/0022-1759(90)90029-U
  33. Kim DM, Byun JN. Effects of antibiotic treatment on the results of nested PCRs for scrub typhus. J Clin Microbiol 2008;46:3465-3466.
  34. Park SM, Ko HJ, Shim DH, Yang JY, Park YH, Curtiss R 3rd, Kweon MN. MyD88 signaling is not essential for induction of antigen-specific B cell responses but is indispensable for protection against Streptococcus pneumoniae infection following oral vaccination with attenuated Salmonella expressing PspA antigen. J Immunol 2008;181:6447-6455. https://doi.org/10.4049/jimmunol.181.9.6447
  35. Shim BS, Park SM, Quan JS, Jere D, Chu H, Song MK, Kim DW, Jang YS, Yang MS, Han SH, et al. Intranasal immunization with plasmid DNA encoding spike protein of SARS-coronavirus/polyethylenimine nanoparticles elicits antigen-specific humoral and cellular immune responses. BMC Immunol 2010;11:65.
  36. Chen CH, Liao WC, Tu CY. Scrub typhus pneumonitis. QJM 2011;104:537-538. https://doi.org/10.1093/qjmed/hcq073
  37. Premaratna R, Loftis AD, Chandrasena TG, Dasch GA, de Silva HJ. Rickettsial infections and their clinical presentations in the Western Province of Sri Lanka: a hospital-based study. Int J Infect Dis 2008;12:198-202. https://doi.org/10.1016/j.ijid.2007.06.009
  38. Fukuyama Y, Okada K, Yamaguchi M, Kiyono H, Mori K, Yuki Y. Nasal administration of cholera toxin as a mucosal adjuvant damages the olfactory system in mice. PLoS One 2015;10:e0139368.
  39. Hanson BA. Effect of immune serum on infectivity of Rickettsia tsutsugamushi. Infect Immun 1983;42:341-349. https://doi.org/10.1128/iai.42.1.341-349.1983
  40. Hauptmann M, Kolbaum J, Lilla S, Wozniak D, Gharaibeh M, Fleischer B, Keller CA. Protective and pathogenic roles of CD8+ T lymphocytes in murine Orientia tsutsugamushi infection. PLoS Negl Trop Dis 2016;10:e0004991.
  41. Jerrells TR, Eisemann CS. Role of T-lymphocytes in production of antibody to antigens of Rickettsia tsutsugamushi and other Rickettsia species. Infect Immun 1983;41:666-674. https://doi.org/10.1128/iai.41.2.666-674.1983
  42. Forbes EK, Sander C, Ronan EO, McShane H, Hill AV, Beverley PC, Tchilian EZ. Multifunctional, high-level cytokine-producing Th1 cells in the lung, but not spleen, correlate with protection against Mycobacterium tuberculosis aerosol challenge in mice. J Immunol 2008;181:4955-4964. https://doi.org/10.4049/jimmunol.181.7.4955
  43. Lindenstrom T, Agger EM, Korsholm KS, Darrah PA, Aagaard C, Seder RA, Rosenkrands I, Andersen P. Tuberculosis subunit vaccination provides long-term protective immunity characterized by multifunctional CD4 memory T cells. J Immunol 2009;182:8047-8055. https://doi.org/10.4049/jimmunol.0801592
  44. Huster KM, Stemberger C, Busch DH. Protective immunity towards intracellular pathogens. Curr Opin Immunol 2006;18:458-464. https://doi.org/10.1016/j.coi.2006.05.008
  45. Neutra MR, Kozlowski PA. Mucosal vaccines: the promise and the challenge. Nat Rev Immunol 2006;6:148-158. https://doi.org/10.1038/nri1777
  46. Peter JV, Sudarsan TI, Prakash JA, Varghese GM. Severe scrub typhus infection: clinical features, diagnostic challenges and management. World J Crit Care Med 2015;4:244-250. https://doi.org/10.5492/wjccm.v4.i3.244
  47. Seong SY, Huh MS, Jang WJ, Park SG, Kim JG, Woo SG, Choi MS, Kim IS, Chang WH. Induction of homologous immune response to Rickettsia tsutsugamushi Boryong with a partial 56-kilodalton recombinant antigen fused with the maltose-binding protein MBP-Bor56. Infect Immun 1997;65:1541-1545. https://doi.org/10.1128/iai.65.4.1541-1545.1997
  48. Nacy CA, Meltzer MS. Macrophages in resistance to rickettsial infection: macrophage activation in vitro for killing of Rickettsia tsutsugamushi. J Immunol 1979;123:2544-2549. https://doi.org/10.4049/jimmunol.123.6.2544
  49. Hanson B. Susceptibility of Rickettsia tsutsugamushi Gilliam to gamma interferon in cultured mouse cells. Infect Immun 1991;59:4125-4133. https://doi.org/10.1128/iai.59.11.4125-4133.1991
  50. Walker DH, Popov VL, Crocquet-Valdes PA, Welsh CJ, Feng HM. Cytokine-induced, nitric oxide-dependent, intracellular antirickettsial activity of mouse endothelial cells. Lab Invest 1997;76:129-138.
  51. Kodama K, Kawamura S, Yasukawa M, Kobayashi Y. Establishment and characterization of a T-cell line specific for Rickettsia tsutsugamushi. Infect Immun 1987;55:2490-2495. https://doi.org/10.1128/iai.55.10.2490-2495.1987
  52. Woolard MD, Hensley LL, Kawula TH, Frelinger JA. Respiratory Francisella tularensis live vaccine strain infection induces Th17 cells and prostaglandin E2, which inhibits generation of gamma interferon-positive T cells. Infect Immun 2008;76:2651-2659. https://doi.org/10.1128/IAI.01412-07
  53. Zhang X, Gao L, Lei L, Zhong Y, Dube P, Berton MT, Arulanandam B, Zhang J, Zhong G. A MyD88-dependent early IL-17 production protects mice against airway infection with the obligate intracellular pathogen Chlamydia muridarum. J Immunol 2009;183:1291-1300. https://doi.org/10.4049/jimmunol.0803075
  54. Mortensen R, Christensen D, Hansen LB, Christensen JP, Andersen P, Dietrich J. Local Th17/IgA immunity correlate with protection against intranasal infection with Streptococcus pyogenes. PLoS One 2017;12:e0175707.
  55. Lindenstrom T, Woodworth J, Dietrich J, Aagaard C, Andersen P, Agger EM. Vaccine-induced th17 cells are maintained long-term postvaccination as a distinct and phenotypically stable memory subset. Infect Immun 2012;80:3533-3544. https://doi.org/10.1128/IAI.00550-12
  56. Valbuena G, Walker DH. Approaches to vaccines against Orientia tsutsugamushi. Front Cell Infect Microbiol 2013;2:170.
  57. Kundin WD, Liu C, Harmon P, Rodina P. Pathogenesis of scrub typhus infection (Rickettsia tsutsugamushi) as studied by immunofluorescence. J Immunol 1964;93:772-781. https://doi.org/10.4049/jimmunol.93.5.772
  58. Shelite TR, Saito TB, Mendell NL, Gong B, Xu G, Soong L, Valbuena G, Bouyer DH, Walker DH. Hematogenously disseminated Orientia tsutsugamushi-infected murine model of scrub typhus [corrected]. PLoS Negl Trop Dis 2014;8:e2966.
  59. Boyaka PN. Inducing mucosal IgA: a challenge for vaccine adjuvants and delivery systems. J Immunol 2017;199:9-16. https://doi.org/10.4049/jimmunol.1601775