1887

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Volume 71, Issue 4

Calcium phosphate adjuvanted nanoparticles of outer membrane proteins of Typhi as a candidate for vaccine development against Typhoid fever No Access

Abstract

The conventional adjuvants used in vaccines have limitations like induction of an imbalanced Th1 and Th2 immune response. To overcome this limitation, novel adjuvants and newer forms of existing adjuvants like calcium phosphate nanoparticles are being tested.

Calcium phosphate adjuvanted outer membrane proteins vaccine may work as an efficient, safe and cost effective vaccine against Typhi.

Our goals were to evaluate the potential of calcium phosphate nanoparticles as an adjuvant using outer membrane proteins (Omps) of Typhi as antigens for immune response, with montanide (commercially available adjuvant) as control, and its toxicity in rats.

Calcium phosphate adjuvanted outer membrane proteins nanoparticles were synthesized and characterized. The efficacy of vaccine formulation in mice and toxicity assay were carried out in rats.

The calcium phosphate nanoparticles varying in size between 20–50 nm had entrapment efficiency of 41.5% and loading capacity of 54%. The calcium phosphate nanoparticle-Omps vaccine formulation (nanoparticle-Omps) induced a strong humoral immune response, which was significantly higher than the control group for the entire period of study. In the montanide-Omps group the initial very high immune response declined steeply and then remained steady. The immune response induced by nanoparticle-Omps did not change appreciably. The cell mediated immune response as measured by lymphocyte proliferation assay and delayed type hypersensitivity test showed a higher response (<0.01) for the nanoparticles-Omps group as compared to montanide-Omps group. The bacterial clearance assay also showed higher clearance in the nanoparticles-Omps group as compared to montanide-Omps group (approx 1.4%). The toxicity analysis in rats showed no difference in the values of toxicity biomarkers and blood chemistry parameters, revealing vaccine formulation was non-toxic in rats.

Calcium phosphate nanoparticles as adjuvant in vaccines is safe, have good encapsulation and loading capacity and induce a strong cell mediated, humoral and protective immune response.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): calcium phosphate nanoparticles , Salmonella Typhi , typhoid fever and vaccine
Funding
This study was supported by the:
  • Department of Biotechnology, Government of West Bengal (Award BT/PR796ADB90/116/2011)
    • Principle Award Recipient: MumteshKumar Saxena
© 2022 The Authors
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2022-04-27
2024-05-05
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References

  1. Saad NJ, Lynch VD, Antillón M, Yang C, Crump JA et al. Seasonal dynamics of typhoid and paratyphoid fever. Sci Rep 2018; 8:1–9 [View Article] [PubMed]
     [Google Scholar]
  2. Das S, Chowdhury R, Pal A, Okamoto K, Das S. Salmonella Typhi outer membrane protein STIV is a potential candidate for vaccine development against typhoid and paratyphoid fever. Immunobiology 2019; 224:371–382 [View Article] [PubMed]
     [Google Scholar]
  3. Britto C, Pollard AJ, Voysey M, Blohmke CJ. An appraisal of the clinical features of pediatric enteric fever: systematic review and meta-analysis of the age-stratified disease occurrence. Clin Infect Dis 2017; 64:1604–1611 [View Article] [PubMed]
     [Google Scholar]
  4. Baker S, Holt KE, Clements ACA, Karkey A, Arjyal A et al. Combined high-resolution genotyping and geospatial analysis reveals modes of endemic urban typhoid fever transmission. Open Biol 2011; 1:110008 [View Article] [PubMed]
     [Google Scholar]
  5. Chandane P, Gandhi A, Bowalekar S. Study of antibiotic susceptibility pattern of Salmonella typhi in children suffering from enteric fever. Ann Trop Med Public Health 2017; 10:440 [View Article]
     [Google Scholar]
  6. Coward C, Restif O, Dybowski R, Grant AJ, Maskell DJ et al. The effects of vaccination and immunity on bacterial infection dynamics in vivo. PLoS Pathog 2014; 10:e1004359 [View Article] [PubMed]
     [Google Scholar]
  7. Fraser A, Paul M, Goldberg E, Acosta CJ, Leibovici L. Typhoid fever vaccines: systematic review and meta-analysis of randomised controlled trials. Vaccine 2007; 25:7848–7857 [View Article] [PubMed]
     [Google Scholar]
  8. Marathe SA, Lahiri A, Negi VD, Chakravortty D. Typhoid fever & vaccine development: a partially answered question. Indian J Med Res 2012; 135:161–169 [PubMed]
     [Google Scholar]
  9. Owais A, Sultana S, Zaman U, Rizvi A, Zaidi AKM. Incidence of typhoid bacteremia in infants and young children in southern coastal Pakistan. Pediatr Infect Dis J 2010; 29:1035–1039 [View Article] [PubMed]
     [Google Scholar]
  10. Hamid N, Jain SK. Characterization of an outer membrane protein of Salmonella enterica serovar typhimurium that confers protection against typhoid. Clin Vaccine Immunol 2008; 15:1461–1471 [View Article] [PubMed]
     [Google Scholar]
  11. Ghosh S, Chakraborty K, Nagaraja T, Basak S, Koley H et al. An adhesion protein of Salmonella enterica serovar Typhi is required for pathogenesis and potential target for vaccine development. Proc Natl Acad Sci U S A 2011; 108:3348–3353 [View Article] [PubMed]
     [Google Scholar]
  12. Carreño JM, Perez-Shibayama C, Gil-Cruz C, Lopez-Macias C, Vernazza P et al. Evolution of Salmonella Typhi outer membrane protein-specific T and B cell responses in humans following oral Ty21a vaccination: A randomized clinical trial. PLoS One 2017; 12:e0178669 [View Article] [PubMed]
     [Google Scholar]
  13. Pérez-Toledo M, Valero-Pacheco N, Pastelin-Palacios R, Gil-Cruz C, Perez-Shibayama C et al. Salmonella Typhi porins OmpC and OmpF are potent adjuvants for T-dependent and T-independent antigens. Front Immunol 2017; 8:230 [View Article] [PubMed]
     [Google Scholar]
  14. Cai W, Kesavan DK, Wan J, Abdelaziz MH, Su Z et al. Bacterial outer membrane vesicles, a potential vaccine candidate in interactions with host cells based. Diagn Pathol 2018; 13:1–2 [View Article] [PubMed]
     [Google Scholar]
  15. Lin Y, Wang X, Huang X, Zhang J, Xia N et al. Calcium phosphate nanoparticles as a new generation vaccine adjuvant. Expert Rev Vaccines 2017; 16:895–906 [View Article] [PubMed]
     [Google Scholar]
  16. Marrack P, McKee AS, Munks MW. Towards an understanding of the adjuvant action of aluminium. Nat Rev Immunol 2009; 9:287–293 [View Article] [PubMed]
     [Google Scholar]
  17. Hayashi M, Aoshi T, Kogai Y, Nomi D, Haseda Y et al. Optimization of physiological properties of hydroxyapatite as a vaccine adjuvant. Vaccine 2016; 34:306–312 [View Article] [PubMed]
     [Google Scholar]
  18. Wang X, Li X, Ito A, Watanabe Y, Sogo Y et al. Rod-shaped and substituted hydroxyapatite nanoparticles stimulating type 1 and 2 cytokine secretion. Colloids Surf B Biointerfaces 2016; 139:10–16 [View Article] [PubMed]
     [Google Scholar]
  19. Joyappa DH, Kumar CA, Banumathi N, Reddy GR, Suryanarayana VVS. Calcium phosphate nanoparticle prepared with foot and mouth disease virus P1-3CD gene construct protects mice and guinea pigs against the challenge virus. Vet Microbiol 2009; 139:58–66 [View Article] [PubMed]
     [Google Scholar]
  20. Koppad S, Raj GD, Gopinath VP, Kirubaharan JJ, Thangavelu A et al. Calcium phosphate coupled Newcastle disease vaccine elicits humoral and cell mediated immune responses in chickens. Res Vet Sci 2011; 91:384–390 [View Article] [PubMed]
     [Google Scholar]
  21. Saeed MI, Omar AR, Hussein MZ, Elkhidir IM, Sekawi Z. Systemic antibody response to nano-size calcium phospate biocompatible adjuvant adsorbed HEV-71 killed vaccine. Clin Exp Vaccine Res 2015; 4:88–98 [View Article] [PubMed]
     [Google Scholar]
  22. Singh Y, Saxena A, Kumar R, Saxena MK. Isolation and Protein Profiling of Outer Membrane Proteins (OMPS) of Salmonella typhi. Int J Curr Microbiol App Sci 2018; 7:2851–2855 [View Article]
     [Google Scholar]
  23. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951; 193:265–275 [View Article] [PubMed]
     [Google Scholar]
  24. He Q, Mitchell A, Morcol T, Bell SJD. Calcium phosphate nanoparticles induce mucosal immunity and protection against herpes simplex virus type 2. Clin Diagn Lab Immunol 2002; 9:1021–1024 [View Article] [PubMed]
     [Google Scholar]
  25. Tamuly S, Kumar Saxena M, Saxena A, Kumar Mishra R, Jha R. Comparative evaluation of humoral immune response generated by calcium phosphate nanoparticle adjuvanted and saponin-adjuvanted recombinant outermembrane protein 87 (Omp87) of Pasteurella multocida (Serotype B:2) in mice. Journal of Nanopharmaceutics and Drug Delivery 2014; 2:80–86 [View Article]
     [Google Scholar]
  26. Wang D, Xuan L, Zhong H, Gong Y, Shi X et al. Incorporation of well-dispersed calcium phosphate nanoparticles into PLGA electrospun nanofibers to enhance the osteogenic induction potential. RSC Adv 2017; 7:23982–23993 [View Article]
     [Google Scholar]
  27. Kalidoss M, Yunus Basha R, Doble M, Sampath Kumar TS. Theranostic calcium phosphate nanoparticles with potential for multimodal imaging and drug delivery. Front Bioeng Biotechnol 2019; 7:126 [View Article] [PubMed]
     [Google Scholar]
  28. Chaudhary JH, Nayak JB, Brahmbhatt MN, Makwana PP. Virulence genes detection of Salmonella serovars isolated from pork and slaughterhouse environment in Ahmedabad, Gujarat. Vet World 2015; 8:121–124 [View Article] [PubMed]
     [Google Scholar]
  29. Bhat NH, Jain SK. Immunogenic evaluation of a recombinant 49-kilodalton outer membrane protein of Salmonella typhi as a candidate for a subunit vaccine against typhoid. J Infect Dis Immun 2010; 2:30–40
     [Google Scholar]
  30. Oliveira AF, Ruas LP, Cardoso SA, Soares SG, Roque-Barreira MC. Vaccination of mice with Salmonella expressing VapA: mucosal and systemic Th1 responses provide protection against Rhodococcus equi infection. PLoS One 2010; 5:e8644 [View Article] [PubMed]
     [Google Scholar]
  31. Das S, Chowdhury R, Ghosh S, Das S. A recombinant protein of Salmonella Typhi induces humoral and cell-mediated immune responses including memory responses. Vaccine 2017; 35:4523–4531 [View Article] [PubMed]
     [Google Scholar]
  32. Wen H, Dan M, Yang Y, Lyu J, Shao A et al. Acute toxicity and genotoxicity of silver nanoparticle in rats. PLoS One 2017; 12:e0185554 [View Article] [PubMed]
     [Google Scholar]
  33. Cai Y, Tang R. Calcium phosphate nanoparticles in biomineralization and biomaterials. J Mater Chem 2008; 18:3775 [View Article]
     [Google Scholar]
  34. Yüksel S, Pekcan M, Puralı N, Esendağlı G, Tavukçuoğlu E et al. Development and in vitro evaluation of a new adjuvant system containing Salmonella Typhi porins and chitosan. Int J Pharm 2020; 578:119129 [View Article] [PubMed]
     [Google Scholar]
  35. Richa J, Anil K, Anjani S, Mamta P, Rajesh K et al. Heterogeneous expression and functional evaluation of in silico characterized recombinant OmpC of Salmonella Typhimurium as a functional poultry vaccine to eradicate zoonotic transmission. Afr J Biotechnol 2015; 14:2862–2870 [View Article]
     [Google Scholar]
  36. Haque S, Swami P, Khan A. S. Typhi derived vaccines and a proposal for outer membrane vesicles (OMVs) as potential vaccine for typhoid fever. Microb Pathog 2021; 158:105082 [View Article] [PubMed]
     [Google Scholar]
  37. Carreño JM, Perez-Shibayama C, Gil-Cruz C, Printz A, Pastelin R et al. PLGA-microencapsulation protects Salmonella typhi outer membrane proteins from acidic degradation and increases their mucosal immunogenicity. Vaccine 2016; 34:4263–4269 [View Article] [PubMed]
     [Google Scholar]
  38. Abdel-Gawad EI, Hassan AI, Awwad SA. Efficiency of calcium phosphate composite nanoparticles in targeting Ehrlich carcinoma cells transplanted in mice. J Adv Res 2016; 7:143–154 [View Article] [PubMed]
     [Google Scholar]
  39. Banik M, Basu T. Calcium phosphate nanoparticles: a study of their synthesis, characterization and mode of interaction with salmon testis DNA. Dalton Trans 2014; 43:3244–3259 [View Article] [PubMed]
     [Google Scholar]
  40. Tang J, Li L, Howard CB, Mahler SM, Huang L et al. Preparation of optimized lipid-coated calcium phosphate nanoparticles for enhanced in vitro gene delivery to breast cancer cells. J Mater Chem B 2015; 3:6805–6812 [View Article] [PubMed]
     [Google Scholar]
  41. Supraja N, Prasad TNVKV, David E. Synthesis, characterization and antimicrobial activity of the micro/nano structured biogenic silver doped calcium phosphate. Appl Nanosci 2015; 6:31–41 [View Article]
     [Google Scholar]
  42. Nopteeranupharp C, Akkarachaneeyakorn K, Songsasaen A. Synthesis of calcium phosphate composite organogels by using emulsion method for dentine occlusion materials. IOP Conf Ser: Mater Sci Eng 2018; 317:012027 [View Article]
     [Google Scholar]
  43. Jones S, Asokanathan C, Kmiec D, Irvine J, Fleck R et al. Protein coated microcrystals formulated with model antigens and modified with calcium phosphate exhibit enhanced phagocytosis and immunogenicity. Vaccine 2014; 32:4234–4242 [View Article]
     [Google Scholar]
  44. Sokolova V, Knuschke T, Kovtun A, Buer J, Epple M et al. The use of calcium phosphate nanoparticles encapsulating Toll-like receptor ligands and the antigen hemagglutinin to induce dendritic cell maturation and T cell activation. Biomaterials 2010; 31:5627–5633 [View Article] [PubMed]
     [Google Scholar]
  45. Dördelmann G, Kozlova D, Karczewski S, Lizio R, Knauer S et al. Calcium phosphate increases the encapsulation efficiency of hydrophilic drugs (proteins, nucleic acids) into poly(d,l-lactide-co-glycolide acid) nanoparticles for intracellular delivery. J Mater Chem B 2014; 2:7250–7259 [View Article] [PubMed]
     [Google Scholar]
  46. Salerno-Gonçalves R, Wahid R, Sztein MB. Immunization of volunteers with Salmonella enterica serovar Typhi strain Ty21a elicits the oligoclonal expansion of CD8+ T cells with predominant Vbeta repertoires. Infect Immun 2005; 73:3521–3530 [View Article] [PubMed]
     [Google Scholar]
  47. Fresnay S, McArthur MA, Magder LS, Darton TC, Jones C et al. Importance of Salmonella Typhi-Responsive CD8+ T Cell Immunity in a Human Typhoid Fever Challenge Model. Front Immunol 2017; 8:208 [View Article] [PubMed]
     [Google Scholar]
  48. Agarwal RK, Porteen K, Dubal ZB, Asha K et al. Evaluation of recombinant outer membrane protein based vaccine against Salmonella Typhimurium in birds. Biologicals 2013; 41:162–168 [View Article] [PubMed]
     [Google Scholar]
  49. Jawale CV, Chaudhari AA, Jeon BW, Nandre RM, Lee JH. Characterization of a novel inactivated Salmonella enterica serovar Enteritidis vaccine candidate generated using a modified cI857/λ PR/gene E expression system. Infect Immun 2012; 80:1502–1509 [View Article] [PubMed]
     [Google Scholar]
  50. Shippy DC, Fadl AA. Immunological characterization of a gidA mutant strain of Salmonella for potential use in a live-attenuated vaccine. BMC Microbiol 2012; 12:1–10 [View Article] [PubMed]
     [Google Scholar]
  51. Takaya A, Yamamoto T, Tokoyoda K. Humoral immunity vs. Salmonella. Front Immunol 2020; 10:3155 [View Article]
     [Google Scholar]
  52. Ortiz V, Isibasi A, García-Ortigoza E, Kumate J. Immunoblot detection of class-specific humoral immune response to outer membrane proteins isolated from Salmonella typhi in humans with typhoid fever. J Clin Microbiol 1989; 27:1640–1645 [View Article]
     [Google Scholar]
  53. Saxena A, Kumar R, Saxena MK. Vaccination with Salmonella Typhi recombinant outer membrane protein 28 induces humoral but non-protective immune response in rabbit. Vet World 2017; 10:946–949 [View Article]
     [Google Scholar]
  54. Yang Y, Wan C, Xu H, Wei H. Identification and characterization of OmpL as a potential vaccine candidate for immune-protection against salmonellosis in mice. Vaccine 2013; 31:2930–2936 [View Article]
     [Google Scholar]
  55. Wahid R, Simon R, Zafar SJ, Levine MM, Sztein MB. Live oral typhoid vaccine Ty21a induces cross-reactive humoral immune responses against Salmonella enterica serovar Paratyphi A and S. Paratyphi B in humans. Clin Vaccine Immunol 2012; 19:825–834 [View Article]
     [Google Scholar]
  56. Pérez C, Calderón GM, Ximénez C, Melendro EI. Human cell-mediated immune responses to antigenic fractions of Salmonella typhi. Immunology 1996; 89:262–267 [View Article] [PubMed]
     [Google Scholar]
  57. Lalk M, Reifenrath J, Angrisani N, Bondarenko A, Seitz J-M et al. Fluoride and calcium-phosphate coated sponges of the magnesium alloy AX30 as bone grafts: A comparative study in rabbits. J Mater Sci Mater Med 2013; 24:417–436 [View Article]
     [Google Scholar]
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Calcium phosphate adjuvanted nanoparticles of outer membrane proteins of Salmonella Typhi as a candidate for vaccine development against Typhoid fever
J. Med. Microbiol. 71, 001529 (2022); https://doi.org/10.1099/jmm.0.001529
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