Applied Microbiology and Biotechnology

, Volume 97, Issue 10, pp 4333–4342 | Cite as

Expression of a hepatitis A virus antigen in Lactococcus lactis and Escherichia coli and evaluation of its immunogenicity

  • Aleš BerlecEmail author
  • Tadej Malovrh
  • Petra Zadravec
  • Andrej Steyer
  • Matjaž Ravnikar
  • Jerica Sabotič
  • Mateja Poljšak-Prijatelj
  • Borut Štrukelj
Biotechnological products and process engineering


An epidemic shift in Hepatitis A virus (HAV) infection has been observed in recent years in rapidly developing countries, with increasing numbers of severe adult cases which has led to renewed interest in vaccination. Our approach in vaccine development uses recombinant expression of the highly immunogenic HAV antigen VP1-P2a in food-grade lactic acid bacterium Lactococcus lactis and in Escherichia coli. We used genetic constructs that enable nisin-controlled expression of the antigen in L. lactis in three different forms: (a) intracellularly, (b) on the bacterial surface and (c) on the bacterial surface fused with the fragment of the E. coli flagellin molecule that can act as a molecular adjuvant. Expression of the two surface forms of the antigen was achieved in L. lactis, and the resulting antigen-displaying bacteria were administered orally to mice. Half the animals in each of the two groups developed specific IgGs, with titers increasing over time and reaching 1:422 without flagellin and 1:320 with flagellin. A much higher titer 1:25,803 was observed with the parenterally administered antigen, which was purified from E. coli. With the latter, a significant mucosal IgA response was also observed. Despite significant titers, the IgGs elicited with oral or parenteral administration could not prevent HAV from infecting cells in a virus neutralization assay, suggesting that the antibodies cannot recognize viral surface epitopes. Nevertheless, orally administered HAV antigen expressed in L. lactis elicited significant systemic humoral immune response showing the feasibility for development of effective HAV vaccine for mucosal delivery.


Hepatitis A Vaccine Oral delivery Lactococcus lactis Immune response 



This study was supported by the Slovenian Research Agency Grant No. P4-0127. The authors are grateful to Prof. Roger Pain for critical reading of the manuscript.

Supplementary material

253_2013_4722_MOESM1_ESM.pdf (68 kb)
ESM 1 (PDF 68 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Aleš Berlec
    • 1
    Email author
  • Tadej Malovrh
    • 2
  • Petra Zadravec
    • 1
    • 3
  • Andrej Steyer
    • 4
  • Matjaž Ravnikar
    • 3
  • Jerica Sabotič
    • 1
  • Mateja Poljšak-Prijatelj
    • 4
  • Borut Štrukelj
    • 1
    • 3
  1. 1.Department of BiotechnologyJožef Stefan InstituteLjubljanaSlovenia
  2. 2.Institute of Microbiology and Parasitology, Veterinary FacultyUniversity of LjubljanaLjubljanaSlovenia
  3. 3.Faculty of PharmacyUniversity of LjubljanaLjubljanaSlovenia
  4. 4.Institute of Microbiology and Immunology, Faculty of MedicineUniversity of LjubljanaLjubljanaSlovenia

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