Abstract
Chimeric virus-like particles (VLPs) were developed as a candidate for allergen-specific immunotherapy. In this study, hepatitis B core antigen (HBcAg) that genetically fused to Chenopodium album polcalcin (Che a 3)–derived peptide was expressed in E. coli BL21, purified, and VLP formation was evaluated using native agarose gel electrophoresis (NAGE) and transmission electron microscopy (TEM). Chimeric HBc VLPs were characterized in terms of their reactivity to IgE, the induction of blocking IgG and allergen-specific IgE, basophil-activating capacity, and Th1-type immune responses. Results from IgE reactivity and basophil activation test showed that chimeric HBc VLPs lack IgE-binding capacity and basophil degranulation activity. Although chimeric HBc VLPs induced the highest level of efficient polcalcin-specific IgG antibody in comparison to those induced by recombinant Che a 3 (rChe a 3) mixed either with HBc VLPs or alum, they triggered the lowest level of polcalcin-specific IgE in mice following immunization. Furthermore, in comparison to the other antigens, chimeric HBc VLPs produced a polcalcin-specific Th1 cell response. Taken together, genetically fusion of allergen derivatives to HBc VLPs, in comparison to a mix of them, may be a more effective way to induce appropriate immune responses in allergen-specific immunotherapy.
Key points
• The insertion of allergen-derived peptide into major insertion region (MIR) of hepatitis B virus core (HBc) antigen resulted in nanoparticles displaying allergen-derived peptide upon its expression in prokaryotic host.
• The resultant VLPs (chimeric HBc VLPs) did not exhibit IgE reactivity with allergic patients’ sera and were not able to degranulate basophils.
• Chimeric HBc VLPs dramatically improved protective IgG antibody response compared with those induced by allergen mixed either with HBc VLPs or alum.
• Chimeric HBc VLPs induced Th1 responses that were counterparts of Th2 responses (allergic).
• Chimeric HBc VLPs increased IgG2a/ IgG1 ratio and the level of IFN-γ compared to those induced by allergen mixed with either HBc VLPs or alum.
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The authors confirm that the data supporting the findings of this study are available within the manuscript.
References
Ai C, Zhang Q, Ding J, Ren C, Wang G, Liu X, Tian F, Zhao J, Zhang H, Chen YQ, Chen W (2015) Suppression of dust mite allergy by mucosal delivery of a hypoallergenic derivative in a mouse model. Appl Microbiol Biotechnol 99(10):4309–4319
Alexander C, Tarzi M, Larche M, Kay A (2005) The effect of Fel d 1-derived T-cell peptides on upper and lower airway outcome measurements in cat-allergic subjects. Allergy 60(10):1269–1274
Anzaghe M, Schülke S, Scheurer S (2018) Virus-like particles as carrier systems to enhance immunomodulation in allergen immunotherapy. Curr Allergy Asthma Rep 18(12):71
Arora U, Tyagi P, Swaminathan S, Khanna N (2012) Chimeric Hepatitis B core antigen virus-like particles displaying the envelope domain III of dengue virus type 2. J Nanobiotechnol 10(1):30
Bannon GA, Cockrell G, Connaughton C, West CM, Helm R, Stanley JS, King N, Rabjohn P, Sampson HA, Burks AW (2001) Engineering, characterization and in vitro efficacy of the major peanut allergens for use in immunotherapy. Int Arch Allergy Immunol 124(1-3):70–72
Barderas R, Villaba M, Batanero E, Pascual CY, Rodríguez R (2003) Role of profilin and polcalcin in chenopod pollen allergy. Allergy Clin Immunol 111(5):1132–1133
Beeh K-M, Kanniess F, Wagner F, Schilder C, Naudts I, Hammann-Haenni A, Willers J, Stocker H, Mueller P, Bachmann MF (2013) The novel TLR-9 agonist QbG10 shows clinical efficacy in persistent allergic asthma. J Allergy Clin Immunol 131(3):866–874
Burks AW, Calderon MA, Casale T, Cox L, Demoly P, Jutel M, Nelson H, Akdis CA (2013) Update on allergy immunotherapy: American Academy of Allergy, Asthma & Immunology/European Academy of Allergy and Clinical Immunology/PRACTALL consensus report. J Allergy Clin Immunol 131(5):1288–1296. e3
Campana R, Valenta R, Vrtala S, Swoboda I, Focke-Tejkl M, Gieras A, Spitzauer S, Valent P, Blatt K, Linhart B (2014) Hypallergenic mosaic antigens and methods of making same. Google Patents
Care IoLARCo, Animals UoL, Resources NIoHDoR (1985) Guide for the care and use of laboratory animals. National Academies, Washington, DC
Curin M, Khaitov M, Karaulov A, Namazova-Baranova L, Campana R, Garib V, Valenta R (2018) Next-generation of allergen-specific immunotherapies: molecular approaches. Curr Allergy Asthma Rep 18(7):39
Ebert EC, Jabri B (2008) Massive interleukin-12-induced interferon-γ production by interleukin-15-stimulated lamina propria lymphocytes followed by down-regulation of the interleukin-12 receptor. J Immunol 124(4):453–460
Engeroff P, Caviezel F, Storni F, Thoms F, Vogel M, Bachmann MF (2018) Allergens displayed on virus-like particles are highly immunogenic but fail to activate human mast cells. Allergy 73(2):341–349
Ghasemi A, Falak R, Mohamadi M, June Maleki S, Assarezadegan M-A, Jafary M (2020) Incorporation of T-cell epitopes from tetanus and diphtheria toxoids into in-silico-designed hypoallergenic vaccine may enhance the protective immune response against allergens. Iran J Basic Med Sci 23(5):636–644
Gomes AC, Flace A, Saudan P, Zabel F, Cabral-Miranda G, Turabi AE, Manolova V, Bachmann MF (2017) Adjusted particle size eliminates the need of linkage of antigen and adjuvants for appropriated T cell responses in virus-like particle-based vaccines. Front Immunol 8:226
Guo J, Zhou A, Sun X, Sha W, Ai K, Pan G, Zhou C, Zhou H, Cong H, He S (2019) Immunogenicity of a virus-like-particle vaccine containing multiple antigenic epitopes of Toxoplasma gondii against acute and chronic Toxoplasmosis in mice. Front Immunol 10:592
Hauser M, Roulias A, Ferreira F, Egger M (2010) Panallergens and their impact on the allergic patient. Allergy, Asthma Clin Immunol 6(1):1–14
Hedlin G, Graff-Lonnevig V, Heilborn H, Lilja G, Norrlind K, Pegelow K, Sundin B, Lowenstein H (1991) Immunotherapy with cat-and dog-dander extracts: V. Effects of 3 years of treatment. J Allergy Clin Immunol 87(5):955–964
Himly M, Wallner M, Zaborsky N, Hauser M, Gadermaier G, Harrer A, Ferreira F (2012) Physicochemical characterization of recombinant allergens and hypoallergenic variants. Arb Paul Ehrlich Inst Bundesinstitut Impfstoffe Biomed Arzneim Langen Hess 97:101–105
Huber J, Ramos H, Gill M, Farrar J (2010) Type I interferon reverses human Th2 commitment and stability by repressing GATA3. j. Immunol 185(2):813–817
Johnson-Weaver BT, McRitchie S, Mercier KA, Pathmasiri W, Sumner SJ, Chan C, Germolec D, Kulis M, Burks AW, Staats HF (2018) Effect of endotoxin and alum adjuvant vaccine on peanut allergy. Allergy Clin Immunol 141(2):791–794. e8
Kanzler H, Barrat FJ, Hessel EM, Coffman RL (2007) Therapeutic targeting of innate immunity with Toll-like receptor agonists and antagonists. Nat Med 13(5):552–559
Klimek L, Willers J, Hammann-Haenni A, Pfaar O, Stocker H, Mueller P, Renner W, Bachmann M (2011) Assessment of clinical efficacy of CYT003-QbG10 in patients with allergic rhinoconjunctivitis: a phase IIb study. Clin Exp Allergy 41(9):1305–1312
Klimek L, Schendzielorz P, Mueller P, Saudan P, Willers J (2013) Immunotherapy of allergic rhinitis: new therapeutic opportunities with virus-like particles filled with CpG motifs. Am J Rhinol Allergy 27(3):206–212
Klimek L, Kündig T, Kramer MF, Guethoff S, Jensen-Jarolim E, Schmidt-Weber CB, Palomares O, Mohsen MO, Jakob T, Bachmann M (2018) Virus-like particles (VLP) in prophylaxis and immunotherapy of allergic diseases. Allergo J Int 27(8):245–255
Komlósi ZI, Kovács N, Sokolowska M, van de Veen W, Akdis M, Akdis CA (2020) Mechanisms of subcutaneous and sublingual aeroallergen immunotherapy: what is new? Immunol Allergy Clin 40(1):1–14
Kulis M, MacQueen I, Li Y, Guo R, Zhong X-P, Burks AW (2012) Pepsinized cashew proteins are hypoallergenic and immunogenic and provide effective immunotherapy in mice with cashew allergy. J Allergy Clin Immunol 130(3):716–723
Kündig TM, Senti G, Schnetzler G, Wolf C, Vavricka BMP, Fulurija A, Hennecke F, Sladko K, Jennings GT, Bachmann MF (2006) Der p 1 peptide on virus-like particles is safe and highly immunogenic in healthy adults. J Allergy Clin Immunol 117(6):1470–1476
Kündig TM, Klimek L, Schendzielorz P, Renner WA, Senti G, Bachmann MF (2015) Is the allergen really needed in allergy immunotherapy? Curr Treat Options Allergy 2(1):72–82
Ledesma A, Barderas R, Westritschnig K, Quiralte J, Pascual C, Valenta R, Villalba M, Rodriguez R (2006) A comparative analysis of the cross-reactivity in the polcalcin family including Syr v 3, a new member from lilac pollen. Allergy 61(4):477–484
Li JJ, Zhao F, Li J (2011) Polyrotaxanes for applications in life science and biotechnology. Appl Microbiol Biotechnol 90(2):427–443
Linhart B, Valenta R (2005) Molecular design of allergy vaccines. Curr Opin Immunol 17(6):646–655
Mohamadi M, Falak R, Mokhtarian K, Khoramizadeh MR, Sadroddiny E, Kardar GA (2016) Identification and characterization of main allergic proteins in cooked wolf herring fish. Iran J Allergy Asthma Immunol 15(5):363–371
Mohammadi M, Mokhtarian K, Kardar GA, Farrokhi S, Sadroddiny E, Khorramizadeh MR, Falak R (2017) Expression of recombinant parvalbumin from wolf-herring fish and determination of its IgE-binding capability. Food Agric Immunol 28(4):573–585
Mohan T, Berman Z, Luo Y, Wang C, Wang S, Compans RW, Wang B-Z (2017) Chimeric virus-like particles containing influenza HA antigen and GPI-CCL28 induce long-lasting mucosal immunity against H3N2 viruses. Sci Rep 7:40226
Mohsen MO, Speiser DE, Knuth A, Bachmann MF (2020) Virus-like particles for vaccination against cancer. Wiley Interdiscip Rev Nanomed Nanobiotechnol 12(1):e1579
Narisety SD, Keet CA (2012) Sublingual vs oral immunotherapy for food allergy. Drugs 72(15):1977–1989
Nouri HR, Varasteh A, Vahedi F, Chamani J, Afsharzadeh D, Sankian M (2012) Constructing a hybrid molecule with low capacity of IgE binding from Chenopodium album pollen allergens. Immunol Lett 144(1-2):67–77
Oldfield WL, Kay AB, Larché M (2001) Allergen-derived T cell peptide-induced late asthmatic reactions precede the induction of antigen-specific hyporesponsiveness in atopic allergic asthmatic subjects. J Immunol 167(3):1734–1739
Oliveira GA, Wetzel K, Calvo-Calle JM, Nussenzweig R, Schmidt A, Birkett A, Dubovsky F, Tierney E, Gleiter CH, Boehmer G (2005) Safety and enhanced immunogenicity of a hepatitis B core particle Plasmodium falciparum malaria vaccine formulated in adjuvant Montanide ISA 720 in a phase I trial. Infect Immun 73(6):3587–3597
Oliveira M, Amorim MI, Ferreira E, Delgado L, Abreu I (2010) Main airborne Ascomycota spores: characterization by culture, spore morphology, ribosomal DNA sequences and enzymatic analysis. Appl Microbiol Biotechnol 86(4):1171–1181
Pajno GB, Cox L, Caminiti L, Ramistella V, Crisafulli G (2014) Oral immunotherapy for treatment of immunoglobulin E-mediated food allergy: the transition to clinical practice. Pediatr Allergy Immunol Pulmonol 27(2):42–50
Pattenden LK, Middelberg AP, Niebert M, Lipin DI (2005) Towards the preparative and large-scale precision manufacture of virus-like particles. Trends Biotechnol 23(10):523–529
Peyret H, Gehin A, Thuenemann EC, Blond D, El Turabi A, Beales L, Clarke D, Gilbert RJ, Fry EE, Stuart DI, Holmes K, Stonehouse NJ, Whelan M, Rosenberg W, Lomonossoff GP, Rowlands DJ (2015) Tandem fusion of hepatitis B core antigen allows assembly of virus-like particles in bacteria and plants with enhanced capacity to accommodate foreign proteins. PLoS One 10(4):e0120751
Pumpens P, Grens E (2001) HBV core particles as a carrier for B cell/T cell epitopes. Intervirology 44(2–3):98–114
Pumpens P, Borisova G, Crowther R, Grens E (1995) Hepatitis B virus core particles as epitope carriers. Intervirology 38(1–2):63–74
Ren C, Zhang Q, Wang G, Ai C, Hu M, Liu X, Tian F, Zhao J, Chen Y, Wang M, Zhang H, Chen W (2014) Modulation of peanut-induced allergic immune responses by oral lactic acid bacteria-based vaccines in mice. Appl Microbiol Biotechnol 98(14):6353–6364
Salem AK (2014) A promising CpG adjuvant-loaded nanoparticle-based vaccine for treatment of dust mite allergies. Immunotherapy 6(11):1161–1163
Sanz M, Sanchez G, Gamboa P, Vila L, Uasuf C, Chazot M, Dieguez I, De Weck A (2001) Allergen-induced basophil activation: CD63 cell expression detected by flow cytometry in patients allergic to Dermatophagoides pteronyssinus and Lolium perenne. Clin Exp Allergy 31(7):1007–1013
Schmitz N, Dietmeier K, Bauer M, Maudrich M, Utzinger S, Muntwiler S, Saudan P, Bachmann MF (2009) Displaying Fel d1 on virus-like particles prevents reactogenicity despite greatly enhanced immunogenicity: a novel therapy for cat allergy. J Exp Med 206(9):1941–1955
Schödel F, Kelly S, Tinge S, Hopkins S, Peterson D, Milich D, Curtiss III R (1996) Hybrid Hepatitis B virus core antigen as a vaccine carrier moiety. Novel strategies in the design and production of vaccines. Springer, pp 15-21
Senti G, Johansen P, Haug S, Bull C, Gottschaller C, Müller P, Pfister T, Maurer P, Bachmann M, Graf N (2009) Use of a-type CpG oligodeoxynucleotides as an adjuvant in allergen-specific immunotherapy in humans: a phase I/IIa clinical trial. Clin Exp Allergy 39(4):562–570
Sharquie IK, Al-Ghouleh A, Fitton P, Clark MR, Armour KL, Sewell HF, Shakib F, Ghaemmaghami AM (2013) An investigation into IgE-facilitated allergen recognition and presentation by human dendritic cells. BMC Immunol 14(1):54
Siskind GW, Paul WE, Benacerraf B (1966) Studies on the effect of the carrier molecule on antihapten antibody synthesis I. Effect of carrier on the nature of the antibody synthesized. J Exp Med 123(4):673–688
Smarr CB, Bryce PJ, Miller SD (2013) Antigen-specific tolerance in immunotherapy of Th2-associated allergic diseases. Crit Rev Immunol 33(5):389–414
Smith MT, Hawes AK, Bundy BC (2013) Reengineering viruses and virus-like particles through chemical functionalization strategies. Curr Opin Biotechnol 24(4):620–626
Sominskaya I, Skrastina D, Petrovskis I, Dishlers A, Berza I, Mihailova M, Jansons J, Akopjana I, Stahovska I, Dreilina D (2013) A VLP library of C-terminally truncated Hepatitis B core proteins: correlation of RNA encapsidation with a Th1/Th2 switch in the immune responses of mice. PLoS One 8(9):e75938
Storni T, Ruedl C, Schwarz K, Schwendener RA, Renner WA, Bachmann MF (2004) Nonmethylated CG motifs packaged into virus-like particles induce protective cytotoxic T cell responses in the absence of systemic side effects. J Immunol 172(3):1777–1785
Storni F, Zeltins A, Balke I, Heath MD, Kramer MF, Skinner MA, Zha L, Roesti E, Engeroff P, Muri L (2020) Vaccine against peanut allergy based on engineered virus-like particles displaying single major peanut allergens. J Allergy Clin Immunol 145(4):1240–1253. e3
Swoboda I, Bugajska-Schretter A, Verdino P, Keller W, Sperr WR, Valent P, Valenta R, Spitzauer S (2002) Recombinant carp parvalbumin, the major cross-reactive fish allergen: a tool for diagnosis and therapy of fish allergy. J Immunol 168(9):4576–4584
Swoboda I, Bugajska-Schretter A, Linhart B, Verdino P, Keller W, Schulmeister U, Sperr WR, Valent P, Peltre G, Quirce S (2007) A recombinant hypoallergenic parvalbumin mutant for immunotherapy of IgE-mediated fish allergy. J Immunol 178(10):6290–6296
Terhune T, Deth R (2018) Aluminum adjuvant-containing vaccines in the context of the hygiene hypothesis: a risk factor for eosinophilia and allergy in a genetically susceptible subpopulation? Int J Environ Res Public Health 15(5):901
Tinghino R, Twardosz A, Barletta B, Puggioni EM, Iacovacci P, Butteroni C, Afferni C, Mari A, Hayek B, Di Felice G (2002) Molecular, structural, and immunologic relationships between different families of recombinant calcium-binding pollen allergens. J Allergy Clin Immunol 109(2):314–320
Valenta R, Kraft D (2002) From allergen structure to new forms of allergen-specific immunotherapy. Curr Opin Immunol 14(6):718–727
Valenta R, Ferreira F, Focke-Tejkl M, Linhart B, Niederberger V, Swoboda I, Vrtala S (2009) From allergen genes to allergy vaccines. Annu Rev Immunol 28:211–241
Verdino P, Barderas R, Villalba M, Westritschnig K, Valenta R, Rodriguez R, Keller W (2008) Three-dimensional structure of the cross-reactive pollen allergen Che a 3: visualizing cross-reactivity on the molecular surfaces of weed, grass, and tree pollen allergens. J Immunol 180(4):2313–2321
Wang W, Chen X, Xue C, Du Y, Lv L, Liu Q, Li X, Ma Y, Shen H, Cao Y (2012) Production and immunogenicity of chimeric virus-like particles containing porcine reproductive and respiratory syndrome virus GP5 protein. Vaccine 30(49):7072–7077
Westritschnig K, Focke M, Verdino P, Goessler W, Keller W, Twardosz A, Mari A, Horak F, Wiedermann U, Hartl A (2004) Generation of an allergy vaccine by disruption of the three-dimensional structure of the cross-reactive calcium-binding allergen, Phl p 7. J Immunol 172(9):5684–5692
Winther L, Arnved J, Malling HJ, Nolte H, Mosbech H (2006) Side-effects of allergen-specific immunotherapy. A prospective multi-centre study. Clin Exp Allergy 36(3):254–260
Xu J, Guo HC, Wei YQ, Dong H, Han SC, Ao D, Sun DH, Wang HM, Cao SZ, Sun SQ (2014) Self-assembly of virus-like particles of canine parvovirus capsid protein expressed from Escherichia coli and application as virus-like particle vaccine. Appl Microbiol Biotechnol 98(8):3529–3538. https://doi.org/10.1007/s00253-013-5485-6
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The Bushehr University of Medical Sciences provided this work a research budget. This study is the MS thesis of the first author and was supported by a research grant (code: 1218) from Bushehr University of Medical Sciences.
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MM, IN, and SJM conceived and designed research. MZ, AB, NM, PD, JA, SH, Kh H, and FO conducted experiments. MM, SJM, and SA contributed new reagents or analytical tools. AM and GR analyzed data. MM and SJM wrote the manuscript. All authors read and approved the manuscript.
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In case animals: All animal dealings in this study were performed under protocol approved by the Animal Care and Use Committee of Bushehr University of Medical Sciences–Iran (Permit number: IR.BPUMS.REC.1397.011). The animal procedures were in accordance with the Specific National Ethical Guidelines for Biomedical Research issued by the Research and Technology Deputy of Ministry of Health and Medicinal Education (MOHME) of Iran (issued in 2005).
In case human: All procedures performed in studies involving human participants were in accordance with the ethical standards of the Ethics Committee, Bushehr University of Medical Sciences with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.
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Sani, M.Z., Bargahi, A., Momenzadeh, N. et al. Genetically engineered fusion of allergen and viral-like particle induces a more effective allergen-specific immune response than a combination of them. Appl Microbiol Biotechnol 105, 77–91 (2021). https://doi.org/10.1007/s00253-020-11012-0
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DOI: https://doi.org/10.1007/s00253-020-11012-0