Abstract
Low pathogenic H9N2 subtype avian influenza virus (AIV) can lead to moderate respiratory symptoms and low egg production rates in poultry. Due to its immunologic suppression, other various infectious pathogens give rise to the co-infection of hosts, causing heavy economic losses in the commercial poultry industry in both China and worldwide. Therefore, it is time to explore a novel, safe, and effective vaccine. We have already made use of the surface of Lactobacillus plantarum to display AIV HA2 (NC8-pSIP409-pgsA′-HA2), which demonstrated that it has a good immunogenicity. In this study, by evaluating the immune protection effect of NC8-pSIP409-pgsA′-HA2 on chickens, we found that the hemagglutination inhibition (HI) antibodies, specificity IgG antibody in chickens, the sIgA titer in broncho alveolar lavage fluids (BALF), and the T cell response were increased notably after oral vaccination with NC8-pSIP409-pgsA′-HA2. In addition, weight loss, lung titers, and lung pathologies were improved when chickens were orally vaccinated with NC8-pSIP409-pgsA′-HA2 after challenge with H9N2 AIV. This strategy lays the foundation for the development of recombinant L. plantarum oral vaccines in the prevention of AIV.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ahmed MS, Jacques LC, Mahallawi W, Ferrara F, Temperton N, Upile N, Vaughan C, Sharma R, Beer H, Hoschler K, McNamara PS, Zhang Q (2015) Cross-reactive immunity against influenza viruses in children and adults following 2009 pandemic H1N1 infection. Antivir Res 114:106–112. https://doi.org/10.1016/j.antiviral.2014.12.008
Anbazhagan K, Sasikumar P, Gomathi S, Priya HP, Selvam GS (2013) In vitro degradation of oxalate by recombinant Lactobacillus plantarum expressing heterologous oxalate decarboxylase. J Appl Microbiol 115(3):880–887. https://doi.org/10.1111/jam.12269
Caggianiello G, Kleerebezem M, Spano G (2016) Exopolysaccharides produced by lactic acid bacteria: from health-promoting benefits to stress tolerance mechanisms. Appl Microbiol Biotechnol 100(9):3877–3886. https://doi.org/10.1007/s00253-016-7471-2
Cai R, Jiang Y, Yang W, Yang W, Shi S, Shi C, Hu J, Gu W, Ye L, Zhou F, Gong Q, Han W, Yang G, Wang C (2016) Surface-displayed IL-10 by recombinant Lactobacillus plantarum reduces Th1 responses of RAW264.7 cells stimulated with poly(I:C) or LPS. J Microbiol Biotechnol 26(2):421–431. https://doi.org/10.4014/jmb.1509.09030
Chanos P, Mygind T (2016) Co-culture-inducible bacteriocin production in lactic acid bacteria. Appl Microbiol Biotechnol 100(10):4297–4308. https://doi.org/10.1007/s00253-016-7486-8
Chen CC, Chiu CH, Lin TY, Shi HN, Walker WA (2009) Effect of probiotics Lactobacillus acidophilus on Citrobacter rodentium colitis: the role of dendritic cells. Pediatr Res 65(2):169–175. https://doi.org/10.1203/PDR.0b013e31818d5a06
Chen HY, Shang YH, Yao HX, Cui BA, Zhang HY, Wang ZX, Wang YD, Chao AJ, Duan TY (2011) Immune responses of chickens inoculated with a recombinant fowlpox vaccine coexpressing HA of H9N2 avain influenza virus and chicken IL-18. Antivir Res 91(1):50–56. https://doi.org/10.1016/j.antiviral.2011.04.007
Dawood MA, Koshio S, Ishikawa M, Yokoyama S (2015) Interaction effects of dietary supplementation of heat-killed Lactobacillus plantarum and beta-glucan on growth performance, digestibility and immune response of juvenile red sea bream, Pagrus major. Fish Shellfish Immunol 45(1):33–42. https://doi.org/10.1016/j.fsi.2015.01.033
Ekiert DC, Friesen RH, Bhabha G, Kwaks T, Jongeneelen M, Yu W, Ophorst C, Cox F, Korse HJ, Brandenburg B, Vogels R, Brakenhoff JP, Kompier R, Koldijk MH, Cornelissen LA, Poon LL, Peiris M, Koudstaal W, Wilson IA, Goudsmit J (2011) A highly conserved neutralizing epitope on group 2 influenza A viruses. Science 333(6044):843–850. https://doi.org/10.1126/science.1204839
Fan X, Hashem AM, Chen Z, Li C, Doyle T, Zhang Y, Yi Y, Farnsworth A, Xu K, Li Z, He R, Li X, Wang J (2015) Targeting the HA2 subunit of influenza A virus hemagglutinin via CD40L provides universal protection against diverse subtypes. Mucosal Immunol 8(1):211–220. https://doi.org/10.1038/mi.2014.59
Haan L, Verweij WR, Holtrop M, Brands R, van Scharrenburg GJ, Palache AM, Agsteribbe E, Wilschut J (2001) Nasal or intramuscular immunization of mice with influenza subunit antigen and the B subunit of Escherichia coli heat-labile toxin induces IgA- or IgG-mediated protective mucosal immunity. Vaccine 19(20–22):2898–2907. https://doi.org/10.1016/S0264-410X(00)00556-9
Higgins JP, Higgins SE, Wolfenden AD, Henderson SN, Torres-Rodriguez A, Vicente JL, Hargis BM, Tellez G (2010) Effect of lactic acid bacteria probiotic culture treatment timing on Salmonella enteritidis in neonatal broilers. Poult Sci 89(2):243–247. https://doi.org/10.3382/ps.2009-00436
Jonges M, van Leuken J, Wouters I, Koch G, Meijer A, Koopmans M (2015) Wind-mediated spread of low-pathogenic avian influenza virus into the environment during outbreaks at commercial poultry farms. PLoS One 10(5):e0125401. https://doi.org/10.1371/journal.pone.0125401
Krammer F, Pica N, Hai R, Tan GS, Palese P (2012) Hemagglutinin stalk-reactive antibodies are boosted following sequential infection with seasonal and pandemic H1N1 influenza virus in mice. J Virol 86(19):10302–10307. https://doi.org/10.1128/jvi.01336-12
Liu D, Shi W, Shi Y, Wang D, Xiao H, Li W, Bi Y, Wu Y, Li X, Yan J, Liu W, Zhao G, Yang W, Wang Y, Ma J, Shu Y, Lei F, Gao GF (2013) Origin and diversity of novel avian influenza A H7N9 viruses causing human infection: phylogenetic, structural, and coalescent analyses. Lancet 381(9881):1926–1932. https://doi.org/10.1016/s0140-6736(13)60938-1
Liu DQ, Ge JW, Qiao XY, Jiang YP, Liu SM, Li YJ (2012) High-level mucosal and systemic immune responses induced by oral administration with Lactobacillus-expressed porcine epidemic diarrhea virus (PEDV) S1 region combined with Lactobacillus-expressed N protein. Appl Microbiol Biotechnol 93(6):2437–2446. https://doi.org/10.1007/s00253-011-3734-0
Margine I, Krammer F, Hai R, Heaton NS, Tan GS, Andrews SA, Runstadler JA, Wilson PC, Albrecht RA, Garcia-Sastre A, Palese P (2013) Hemagglutinin stalk-based universal vaccine constructs protect against group 2 influenza A viruses. J Virol 87(19):10435–10446. https://doi.org/10.1128/jvi.01715-13
Narita J, Okano K, Tateno T, Tanino T, Sewaki T, Sung MH, Fukuda H, Kondo A (2006) Display of active enzymes on the cell surface of Escherichia coli using PgsA anchor protein and their application to bioconversion. Appl Microbiol Biotechnol 70(5):564–572. https://doi.org/10.1007/s00253-005-0111-x
Peng Y, Xie ZX, Liu JB, Pang YS, Deng XW, Xie ZQ, Xie LJ, Fan Q, Luo SS (2013) Epidemiological surveillance of low pathogenic avian influenza virus (LPAIV) from poultry in Guangxi Province, Southern China. PLoS One 8(10):e77132. https://doi.org/10.1371/journal.pone.0077132
Poo H, Pyo HM, Lee TY, Yoon SW, Lee JS, Kim CJ, Sung MH, Lee SH (2006) Oral administration of human papillomavirus type 16 E7 displayed on Lactobacillus casei induces E7-specific antitumor effects in C57/BL6 mice. Int J Cancer 119(7):1702–1709. https://doi.org/10.1002/ijc.22035
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. https://doi.org/10.1007/s00253-014-5678-7
Shi SH, Yang WT, Yang GL, Cong YL, Huang HB, Wang Q, Cai RP, Ye LP, JT H, Zhou JY, Wang CF, Li Y (2014) Immunoprotection against influenza virus H9N2 by the oral administration of recombinant Lactobacillus plantarum NC8 expressing hemagglutinin in BALB/c mice. Virology 464-465:166–176. https://doi.org/10.1016/j.virol.2014.07.011
Shi SH, Yang WT, Yang GL, Zhang XK, Liu YY, Zhang LJ, Ye LP, JT H, Xing X, Qi C, Li Y, Wang CF (2016) Lactobacillus plantarum vaccine vector expressing hemagglutinin provides protection against H9N2 challenge infection. Virus Res 211:46–57. https://doi.org/10.1016/j.virusres.2015.09.005
Stanekova Z, Adkins I, Kosova M, Janulikova J, Sebo P, Vareckova E (2013) Heterosubtypic protection against influenza A induced by adenylate cyclase toxoids delivering conserved HA2 subunit of hemagglutinin. Antivir Res 97(1):24–35. https://doi.org/10.1016/j.antiviral.2012.09.008
Stanekova Z, Vareckova E (2010) Conserved epitopes of influenza A virus inducing protective immunity and their prospects for universal vaccine development. Virol J 7:351. https://doi.org/10.1186/1743-422x-7-351
Tutykhina IL, Sedova ES, Gribova IY, Ivanova TI, Vasilev LA, Rutovskaya MV, Lysenko AA, Shmarov MM, Logunov DY, Naroditsky BS, Tillib SV, Gintsburg AL (2013) Passive immunization with a recombinant adenovirus expressing an HA (H5)-specific single-domain antibody protects mice from lethal influenza infection. Antivir Res 97(3):318–328. https://doi.org/10.1016/j.antiviral.2012.12.021
Wang Q, Wang ZN, Huang KY, Yao XR, Wang CY, Yang GL, Jiang YL, Wang CF (2017) Construction of novel functional lactic acid bacteria with surface displayed HA2 protein of H9N2 subtype AIV. Chinese. Vet Sci 47(04):476–480. 10.16656/j.issn.1673-4696.2017.04.012 (In Chinese)
Wyszynska A, Kobierecka P, Bardowski J, Jagusztyn-Krynicka EK (2015) Erratum to: lactic acid bacteria--20 years exploring their potential as live vectors for mucosal vaccination. Appl Microbiol Biotechnol 99(10):4531. https://doi.org/10.1007/s00253-015-6569-2
Yang WT, Yang GL, Shi SH, Liu YY, Huang HB, Jiang YL, Wang JZ, Shi CW, Jing YB, Wang CF (2017a) Protection of chickens against H9N2 avian influenza virus challenge with recombinant Lactobacillus plantarum expressing conserved antigens. Appl Microbiol Biotechnol 101(11):4593–4603. https://doi.org/10.1007/s00253-017-8230-8
Yang WT, Yang GL, Wang Q, Huang HB, Jiang YL, Shi CW, Wang JZ, Huang KY, Jin YB, Wang CF (2017b) Protective efficacy of Fc targeting conserved influenza virus M2e antigen expressed by Lactobacillus plantarum. Antivir Res 138:9–21. https://doi.org/10.1016/j.antiviral.2016.11.025
Yang ZF, Mok CK, Peiris JS, Zhong NS (2015) Human infection with a novel avian influenza A (H5N6) virus. N Engl J Med 373(5):487–489. https://doi.org/10.1056/NEJMc1502983
Yoon SW, Lee TY, Kim SJ, Lee IH, Sung MH, Park JS, Poo H (2012) Oral administration of HPV-16 L2 displayed on Lactobacillus casei induces systematic and mucosal cross-neutralizing effects in Balb/c mice. Vaccine 30(22):3286–3294. https://doi.org/10.1016/j.vaccine.2012.03.009
Zhou J, Wang D, Gao R, Zhao B, Song J, Qi X, Zhang Y, Shi Y, Yang L, Zhu W, Bai T, Qin K, Lan Y, Zou S, Guo J, Dong J, Dong L, Zhang Y, Wei H, Li X, Lu J, Liu L, Zhao X, Li X, Huang W, Wen L, Bo H, Xin L, Chen Y, Xu C, Pei Y, Yang Y, Zhang X, Wang S, Feng Z, Han J, Yang W, Gao GF, Wu G, Li D, Wang Y, Shu Y (2013) Biological features of novel avian influenza A (H7N9) virus. Nature 499(7459):500–503. https://doi.org/10.1038/nature12379
Acknowledgments
This work was supported by the National Key Research and Development Program of China (2017YFD0501000), the National High-Tech R&D Program of China (863 program) (2013AA102806), National Natural Science Foundation of China (31672528, 31272541), Science and Technology Development Program of Jilin Province (20160519011JH, 20170204034NY), Special Funds for Industrial Innovation of Jilin Province (2016C063), “Thirteen Five-year Plan” for Sci & Tech Research Program of Jilin Education Department of P.R. China (JJKH20170318KJ), the Doctoral Project sponsored by the Scientific Research Foundation of Jilin Agricultural University of China (201601), and the Undergraduate Innovative Training Program of Jilin Province (301).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that there are no competing interests.
Ethical approval
All applicable international and national guidelines for the care and use of chickens were followed.
Rights and permissions
About this article
Cite this article
Yang, WT., Yang, GL., Yang, X. et al. Recombinant Lactobacillus plantarum expressing HA2 antigen elicits protective immunity against H9N2 avian influenza virus in chickens. Appl Microbiol Biotechnol 101, 8475–8484 (2017). https://doi.org/10.1007/s00253-017-8600-2
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-017-8600-2