Expression of avian influenza haemagglutinin (H5) and chicken interleukin 2 (chIL-2) under control of the ptcB promoter in Lactococcus lactis

• Authors: Katarzyna Szatraj , Agnieszka Szczepankowska , Violetta Sączyńska , Katarzyna Florys , Beata Gromadzka , Krzysztof Łepek , Grażyna Płucienniczak , Bogusław Szewczyk , Włodzimierz Zagórski-Ostoja , Jacek Bardowski
• Languages of publication: EN

Abstracts

EN

Gram-positive and nonpathogenic lactic acid bacteria (LAB) are considered to be promising candidates for the development of new, safe systems of heterologous protein expression. Recombinant LAB has been shown to induce specific local and systemic immune response against selected pathogens, and could be a good alternative to classical attenuated carriers. The main goal of our study was to express the avian influenza haemagglutinin (H5) and chicken interleukin 2 (chIL-2) in Lactococcus lactis. Results of this study were anticipated to lead to construction of lactococcal strain(s) with potential vaccine properties against the avian influenza A (H5N1) virus. Expression of the cloned H5 gene, its His-tagged variant and chIL-2 gene, under the control of the ptcB gene promoter was attested by RT-PCR on transcriptional level and Western or dot blot analysis on translational level, demonstrating that system can be an attractive solution for production of heterologous proteins. The results of the preliminary animal trial conducted in mice are a promising step toward development of a vaccine against avian bird flu using Lactococcus lactis cells as antigen carriers.

Keywords

EN

Lactococcus lactis; ptcB promoter; heterologous gene expression; avian influenza H5N1; H5 haemagglutinin; chicken interleukin-2

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2014
• Volume: 61
• Issue: 3
• Pages: 609-614

Dates

published

2014

received

2014-05-30

revised

2014-09-08

accepted

2014-09-10

(unknown)

2014-09-18

Contributors

author

Katarzyna Szatraj

• Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland

author

Agnieszka Szczepankowska

• Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland

author

Violetta Sączyńska

• Institute of Biotechnology and Antibiotics, Department of Bioengineering, Warsaw, Poland

author

Katarzyna Florys

• Institute of Biotechnology and Antibiotics, Department of Bioengineering, Warsaw, Poland

author

Beata Gromadzka

• University of Gdansk and Medical University of Gdańsk, Intercollegiate Faculty of Biotechnology, Department of Recombinant Vaccines, Gdańsk, Poland

author

Krzysztof Łepek

• University of Gdansk and Medical University of Gdańsk, Intercollegiate Faculty of Biotechnology, Department of Recombinant Vaccines, Gdańsk, Poland

author

Grażyna Płucienniczak

• Institute of Biotechnology and Antibiotics, Department of Bioengineering, Warsaw, Poland

author

Bogusław Szewczyk

• University of Gdańsk, Institute of Biotechnology, Department of Molecular Virology, Gdańsk, Poland

author

Włodzimierz Zagórski-Ostoja

• Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland

author

Jacek Bardowski

• Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland

References

• Aleksandrzak-Piekarczyk T, Kok J, Renault P, Bardowski J (2005) Alternative lactose catabolic pathway in Lactococcus lactis IL1403. Appl Environ Microbiol 71: 6060-6069.
• Aleksandrzak-Piekarczyk T, Polak J, Jezierska B, Renault P, Bardowski J (2011) Genetic characterization of the CcpA-dependent, cellobiose-specific PTS system comprising CelB, PtcB and PtcA that transports lactose in Lactococcus lactis IL1403. Int J Food Microbiol 145: 186-194.
• Bermúdez-Humarán LG, Langella P, Miyoshi A, Gruss A, Guerra RT, Montes de Oca-Luna R, Le Loir Y (2002) Production of Human Papillomavirus Type 16 E7 Protein in Lactococcus lactis. Appl Environ Microbiol 68: 917-922.
• Chopin A, Chopin MC, Moillo-Batt A, Langella P (1984) Two plasmid-determined restriction and modification systems in Streptococcus lactis. Plasmid 11: 260-263.
• Drouault S, Corthier G, Ehrlich SD, Renault P (1999) Survival, physiology, and lysis of Lactococcus lactis in the digestive tract. Appl Environ Microbiol 65: 4881-4886.
• Enouf V, Langella P, Commissaire J, Cohen J, Corthier G (2001) Bovine rotavirus nonstructural protein 4 produced by Lactococcus lactis is antigenic and immunogenic. Appl Environ Microbiol 67: 1423-1428.
• Gasson MJ, de Vos WM. (1994) Genetics and biotechnology of lactic acid bacteria. Blackie Academic and Professional, Glasgow.
• Geoffroy MC, Guyard C, Quatannens B, Pavan S, Lange M, Mercenier A (2000) Use of green fluorescent protein to tag lactic acid bacterium strains under development as live vaccine vectors. Appl Environ Microbiol 66: 383-391.
• Guillemin N, Meunier B, Jurie C, Cassar-Malek I, Hocquette JF, Leveziel H, Picard B (2009) Validation of a dot-blot quantitative technique for large scale analysis of beef tenderness biomarkers. J Physiol Pharmacol 60: 91-97.
• Holo H, Nes IF (1989) High-frequency transformation, by electroporation, of Lactococcus lactis subsp. cremoris grown with glycine in osmotically stabilized media. Appl Environ Microbiol 55: 3119-3123.
• Lamm ME (1997) Interaction of antigens and antibodies at mucosal surfaces. Annu Rev Microbiol 51: 311-340.
• Le Loir Y, Gruss A, Ehrlich SD, Langella P (1998) A nine-residue synthetic propeptide enhances secretion efficiency of heterologous proteins in Lactococcus lactis. J Bacteriol 180: 1895-1903.
• Lillehoj HS, Min W, Choi KD, Babu US, Burnside J, Miyamoto T, Rosenthal BM, Lillehoj LP (2001) Molecular, cellular, and functional characterization of chicken cytokines homologous to mammalian IL-15 and IL-2. Vet Immunol Immunopathol 82: 229-244.
• Mota RM, Moreira JL, Souza MR, Horta MF, Teixeira SM, Neumann E, Nicoli JR, Nunes AC (2006) Genetic transformation of novel isolates of chicken Lactobacillus bearing probiotic features for expression of heterologous proteins: a tool to develop live oral vaccines. BMC Biotechnology 6: 2.
• Perdigon G, Fuller R, Raya R (2001) Lactic acid bacteria and their effect on the immune system. Curr Issues Intest Microbiol 2: 27-42.
• Ramasamy R, Yasawardena S, Zomer A, Venema G, Kok J, Leenhouts K (2006) Immunogenicity of a malaria parasite antigen displayed by Lactococcus lactis in oral immunisations. Vaccine 24: 3900-3908.
• Ribeiro LA, Azevedo V, Le Loir Y, Oliveira SC, Dieye Y, Piard JC, Gruss A, Langella P (2002) Production and targeting of the Brucella abortus antigen L7/L12 in Lactococcus lactis: a first step towards food-grade live vaccines against brucellosis. Appl Environ Microbiol 68: 910-916.
• Sánchez J, Borrero J, Gómez-Sala B, Basanta A, Herranz C, Cintas LM, Hernández PE (2008) Cloning and heterologous production of hiracin JM79, a Sec-dependent bacteriocin produced by Enterococcus hirae DCH5, in lactic acid bacteria and Pichia pastoris. Appl Environ Microbiol 74: 2471-2479.
• Staats HF, Jackson RJ, Marinaro M, Takahashi I, Kiyono H, McGhee JR (1994) Mucosal immunity to infection with implications for vaccine development. Curr Opin Immunol 6: 572-583.
• Terzaghi E, Sandine WE (1975) Improved medium for lactic streptococci and their bacteriophages. Appl Environ Microbiol 29: 807-813.
• Titgemeyer F, Hillen W (2002) Global control of sugar metabolism: a gram-positive solution. Antonie Van Leeuwenhoek 82: 59-71.
• Underdown BJ, Mestecky J (1994) Mucosal Immunoglobulin. In Handbook of Mucosal Immunology, Ogra P, Mesteck J, Lamm ME, Strober W, McGhee JR, eds, pp 79-98. Academic Press, Boston, MA.
• Wegmann U, O'Connell-Motherway M, Zomer A, Buist G, Shearman C, Canchaya C, Ventura M, Goesmann A, Gasson MJ, Kuipers OP, van Sinderen D, Kok J (2007) Complete genome sequence of the prototype lactic acid bacterium Lactococcus lactis subsp. cremoris MG1363. J Bacteriol 189: 3256-3270.
• Wells JM, Wilson PW, Norton PM, Gasson JM, Le Page RW (1993) Lactococcus lactis: high-level expression of tetanus toxin fragment C and protection against lethal challenge. Mol Microbiol 8: 1155-1162.
• Wells JM, Mercenier A (2008) Mucosal delivery of therapeutic and prophylactic molecules using lactic acid bacteria. Nat Rev Microbiol 6: 349-362.
• Wood BJB, Warner PJ (2003) Genetics of lactic acid bacteria. New York: Kluwer Academic/Plenum.
• Xin KQ, Hoshino Y, Toda Y, Igimi S, Kojima Y, Jounai N, Ohba K, Kushiro A, Kiwaki M, Hamajima K, Klinman D, Okuda K (2003) Immunogenicity and protective efficacy of orally administered recombinant Lactococcus lactis expressing surface-bound HIV. Env Blood 102: 223–228.