Mucosal Immunization with Recombinant Lactococcus lactis

  • Lisa Chamberlain
  • Jeremy M. Wells
  • Karen Robinson
  • Karin Schofield
  • Richard Le Page
Part of the Biotechnology Intelligence Unit book series (BIOIU)


The United States National Institutes of Health has recently pointed out that despite many years of intensive vaccine research and the development of sophisticated effective vaccines, thousands of people, mainly children, die each year as a result of vaccine-preventable diseases (Jordan Report 1995). A significant factor contributing to this mortality rate is the problem of implementing vaccination programs on a large scale due to the need for trained personnel to administer parenteral inoculations, repeat visits for booster inoculations, and concern over needle stick injuries. The scale of the problem is large, with a projected 125 million children requiring immunization annually by the year 2000. These problems have generated a great deal of interest in the development of new vaccines which could be mucosally administered, introducing the possibility of self-administration of booster inoculations. This would increase the probability of repeated doses being administered and decrease the need for trained personnel, and so decrease costs.


Lactic Acid Bacterium Mucosal Immunization Lactococcus Lactis Schistosoma Mansoni Booster Inoculation 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


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  1. 1.
    Mestecky J, Eldridge JH. Targeting and controlled release of antigens for the effective induction of secretory antibody responses. Current Opinion in Immunology 1991; 3:492–495.CrossRefGoogle Scholar
  2. 2.
    Challacombe SJ, Rahman D, Jeffery H et al. Enhanced secretory IgA and systemic IgG antibody responses after oral immunization with biodegradable microparticles containing antigen. Immunol 1992; 76:164–168.Google Scholar
  3. 3.
    Eldridge JH, Hammond CJ, Meulbroek JA et al. Controlled vaccine release in the gut-associated lymphoid tissues. 1. Orally administered biodegradable microspheres target the Peyer’s patches. Journal of Controlled Release 1990; 11:205–214.CrossRefGoogle Scholar
  4. 4.
    Vadolas J, Davies JK, Wright PJ et al. Intranasal immunization with liposomes induces strong mucosal immune responses in mice. Eur J Immunol 1995; 25:969–975.CrossRefGoogle Scholar
  5. 5.
    Challacombe SJ. Salivary antibodies and systemic tolerance in mice after oral immunization with bacterial antigens. Annals of the New York Academy of Sciences. 1983; 177–193.Google Scholar
  6. 6.
    Chatfield SN, Dougan G, Roberts M et al. Progress in the development of multivalent oral vaccines based on live attenuated Salmonella. In: Kurstak E, ed. Modern Vaccinology, New York: Plenum Press 1994: 55–86.CrossRefGoogle Scholar
  7. 7.
    Stover CK, de la Cruz VF, Fuerst et al. New use of BCG for recombinant vaccines. Nature 1991; 351:456–460.CrossRefGoogle Scholar
  8. 8.
    Roberts M, Chatfield SN, Dougan G et al. Salmonella as carriers of heterologous antigens In: O’Hagan DT ed. Novel Delivery Systems for Oral Vaccines. Florida: CRC Press 1994:27–58.Google Scholar
  9. 9.
    Wells JM, Norton PM, Le Page RWF. Progress in the development of mucosal vaccines based on Lactococcus lactis. Int Dairy Journal 1995; 5:1071–1079.CrossRefGoogle Scholar
  10. 10.
    Gruzza M, Fons M, Ouriet MF et al. Study of gene transfer in vitro and in the digestive tract of gnotobiotic mice from Lactococcus lactis strains to various strains belonging to human intestinal flora. Microb Releases 1994; 2:183–189.Google Scholar
  11. 11.
    Klijn N, Weerkamp AH, de Vos WM. Genetic marking of Lactococcus lactis shows its survival in the human gastrointestinal tract. Appl Environ Microbiol 1995; 61:2771–2774.Google Scholar
  12. 12.
    Eldridge JH, Gilley RM, Staas JK et al. Biodegradable microspheres: vaccine delivery system for oral immunization. Curr Topics in Microbiol and Immunol. 1989; 146:59–66.CrossRefGoogle Scholar
  13. 13.
    Eldridge JH, Hammond CJ, Meulbroek JA et al. J Cont Release 1990; 11:205–214.CrossRefGoogle Scholar
  14. 14.
    Dahlgren UIH, Wold AE, Hanson LA et al. Expression of a dietary protein in E. coli renders it strongly antigenic to gut lymphoid tissue. Immunol. 1991; 73:394.Google Scholar
  15. 15.
    Cox DS, and Taubman MA. Oral induction of the secretory antibody response by soluble and particulate antigens. Int Arch Allergy Appl Immun 1984;74:249.CrossRefGoogle Scholar
  16. 16.
    Norton PM, Brown HWG, Le Page RWF. The immune response to Lactococcus lactis: implications for its use as a delivery vehicle. FEMS Microbiol Lett 1994; 120:249–256.CrossRefGoogle Scholar
  17. 17.
    Robinson K, Chamberlain LM, Collingwood KM et al. Oral and Intra nasal vaccination with recombinant Lactococcus lactis can induce high level immune responses to tetanus toxin fragment C elicits. Joint Congress of the British and Netherlands Societies for Immunology, Brighton, England. Immunol 1995; 86 (supply):27.Google Scholar
  18. 18.
    Schödel F, Kelly SM, Peterson DI et al. Hybrid hepatitis B virus core-pre-S proteins synthesized in avirulent Salmonella typhimurium and Salmonella typhi for oral vaccination. Infect Immun 1994; 62:1669–1676.Google Scholar
  19. 19.
    Wells JM, Robinson K, Chamberlain LM et al. Lactic acid bacteria as vaccine delivery vehicles, Antonie van Leeuwenhoek, 1996; 70:317–330.CrossRefGoogle Scholar
  20. 20.
    Wells JM, Wilson PW, Norton PM et al. Lactococcus lactis: high level expression of tetanus toxin fragment C and protection against lethal challenge. Mol Microbiol 1993; 8:1156–1162.CrossRefGoogle Scholar
  21. 21.
    Studier FW, Rosenberg AH, Dunn JJ et al. Use of T7 RNA polymerase to direct expression of cloned genes. Meth Enzymol 1990; 185:60–89.CrossRefGoogle Scholar
  22. 22.
    de Vos WM, Boerrigter I, van Rooyen RJ et al. Characterization of the lactose-specific enzymes of the phosphotransferase system in Lactococcus lactis. J Biol Chem 1990; 265:22554–2259.Google Scholar
  23. 23.
    van Rooijen RJ, Gasson MJ, de Vos WM. Characterization of the promoter of the Lactococcus lactis lactose operon: contribution of flanking sequences and LacR repressor to its activity. J Bacteriol 1992; 1745:2273–2280.Google Scholar
  24. 24.
    van Rooijen RJ, de Vos WM. Purification of the Lactococcus lactis LacR repressor and characterization of its DNA binding sites. RJ Roojen, PhD thesis 1993; pp101–119.Google Scholar
  25. 25.
    Wells JM, Wilson PW, Norton PM et al. A model system for the investigation of heterologous protein protein secretion pathways in Lactococcus lactis. Appl Environ Microbiol 1993; 59:3934–3959.Google Scholar
  26. 26.
    Wells JM, Schofield KM. Cloning and expression vectors for lactococci. In: Bozoglu F, Ray B eds. Lactic Acid Bacteria: Current Advances in Metabolism, Genetics, and Applications. NATO ASI Series Vol H 98. Heidelberg: Springer-Verlag, 1996: 37–62.Google Scholar
  27. 27.
    Simmons LC, Yansura DG. Translational level is a critical factor for the secretion of heterologous proteins in Escherichia coli. Nature Biotechnology 1996; 14:629–634.CrossRefGoogle Scholar
  28. 28.
    Ishida M, Oshima T. A leader open reading frame is essential for the expression in E. coli of GC-rich leuB gene of an extreme thermophile, Thermus thermophilus. FEMS Microbiol Lett 1996; 135:137–142.CrossRefGoogle Scholar
  29. 29.
    Brantl S, Behnke D. Characterisation of the minimal origin required for replication of the streptococcal plasmid PIP501 in Bacillus subtilis. Mol Microbiol 1992; 6:3501–3510.CrossRefGoogle Scholar
  30. 30.
    Gruss A, Ehrlich SD. The family of highly interrelated single-stranded deoxyribonucleic acid plasmids. Microbiol Rev 1989; 53:231–241.Google Scholar
  31. 31.
    Waterfield NR, Le Page RWF, Wells JM. The isolation of lactococcal promoters and their use in investigating bacterial luciferase synthesis in Lactococcus lactis. Gene 1995; 165:9–15.CrossRefGoogle Scholar
  32. 32.
    Hildebrand GD. Investigation of Cholera toxin B subunit expression in Lactococcus lactis. M.Sc. thesis Cambridge, 1993.Google Scholar
  33. 33.
    Schofield KM, Wilson PW, Le Page RWF et al. Optimisation of translation initiation in Lactococcus lactis. 1996; Abstract: Fifth Symposium on Lactic Acid Bacteria, The Netherlands.Google Scholar
  34. 34.
    van de Guchte M, van der Lende T, Kok J et al. A possible contribution of mRNA secondary structure to translation coupling and interference in Lactococcus lactis. FEMS Microbiol Lett 1991; 81:201–208.CrossRefGoogle Scholar
  35. 35.
    Halpern JL, Habig WH, Neale EA et al. Cloning and expression of functional fragment C of tetanus toxin. Infect Immun 1990; 58:1004–1009.Google Scholar
  36. 36.
    Fairweather NF et al, Immunization of mice against tetanus toxin with fragments of tetanus toxin synthesised in E. coli. Infect Immun 1987; 55:2541–2545Google Scholar
  37. 37.
    Norton PM, Brown HWG, Wells JM et al. Factors affecting the immunogenicity of tetanus toxin fragment C expressed in Lactococcus lactis. FEMS Immunol Med Microbiol 1996; 14:167–177.CrossRefGoogle Scholar
  38. 38.
    Norton PM, Wells JM, Brown HWG et al. Protection against tetanus toxin in mice nasally immunized with recombinant Lactococcus lactis expressing TTFC. Vaccine 1996; in press.Google Scholar
  39. 39.
    Coutelier JP, Van der Logt JTM, Heesen FWA et al. IgG2a restriction of murine antibodies elicited by viral infections. J Exp Med 1987; 165: 64–69.CrossRefGoogle Scholar
  40. 40.
    Robinson K, Chamberlain LM, Schofield KM et al. Oral vaccination of mice with recombinant Lactococcus lactis expressing tetanus toxin fragment C elicits both secretory and protective high level systemic immune responses. 1995; Abstract: European Commission Biotechnology Program Conference on Lactic Acid bacteria, Cork, Ireland.Google Scholar
  41. 41.
    Nash AD, Lofthouse SA, Barcham GJ et al. Recombinant cytokines as immunological adjuvants. Immunol and Cell Biol 1993; 71:367–379.CrossRefGoogle Scholar
  42. 42.
    Murray PJ, Aldovini A, Young RA. Manipulation and potentiation of antimycobacterial immunity using recombinant bacille Calmette-Guerin strains that secrete cytokines. Proc Natl Acad Sci USA 1996; 93:934–939.CrossRefGoogle Scholar
  43. 43.
    Capron A, Riveau G, Grzych JM et al. Development of a vaccine strategy against human and bovine schistosomiasis. Background and update. Mem. Inst. Oswaldo Cruz, Rio de Janeiro, 1995; 90:235–240.CrossRefGoogle Scholar
  44. 44.
    Khan CMA, Villareal-Ramos B, Pierce RJ et al. Construction, expression and immunogenicity of multiple tandem copies of the Schistosoma mansoni peptide 115–131 of the P28 glutathione S-trans-ferase expressed as C-terminal fusions to tetanus toxin fragment C in a live aro-attenuated vaccine strain of Salmonella. J Immunol 1994; 153:5635–5642.Google Scholar
  45. 45.
    Khan CMA, Villareal-Ramos B, Pierce RJ et al. Construction, expression and immunogenicity of the Schistosoma mansoni P28 Glutathione S-transferase as a genetic fusion to tetanus toxin fragment C in a live aro-attenuated vaccine strain of Salmonella. Proc Natl Acad Sci USA 1994; 91:11261–11265.CrossRefGoogle Scholar
  46. 46.
    Chamberlain LM, Macpherson A, Wells JM et al. Expression of Schistosoma mansoni P28 antigen in Lactococcus lactis. 1995; Abstract: International Dairy Lactic Acid Bacteria Conference, Palmerston North, New Zealand.Google Scholar
  47. 47.
    Grzych JM, Grezel D, Xu CB et al. IgA antibodies to a protective antigen in human Schistosoma mansoni. J Immunol 1993; 150: 527–535.Google Scholar
  48. 48.
    Chamberlain LM, Robinson K, Wells JM et al. Immune response to whole glutathione S-transferase (P28) and its epitopes from Schistosoma mansoni expressed in Lactococcus lactis. 1995; Abstract: European Commission Biotechnology Program Conference on Lactic Acid Bacteria, Cork, Ireland.Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1997

Authors and Affiliations

  • Lisa Chamberlain
  • Jeremy M. Wells
  • Karen Robinson
  • Karin Schofield
  • Richard Le Page

There are no affiliations available

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