Instruments for oral disease-intervention strategies: recombinant Lactobacillus casei expressing tetanus toxin fragment C for vaccination or myelin proteins for oral tolerance induction in multiple sclerosis.
Lactobacillus strains possess properties that make them attractive candidates as vehicles for oral administration of therapeutics. In this report we describe the construction and analysis of recombinant Lactobacillus casei applicable in oral vaccination against an infectious disease (tetanus) and in oral tolerance induction for intervention in an autoimmune disease, multiple sclerosis. Recombinant L. casei which express surface-anchored tetanus toxin fragment C (TTFC) were generated. Quantitative analysis by flow cytometry demonstrated a high level of cell wall-bound expression of TTFC and immunogenicity was demonstrated by parenteral immunization with whole cell extracts of the recombinants. A series of expression vectors was constructed to secrete human myelin basic protein (hMBP) or hMBP as a fusion protein with beta-glucuronidase from Escherichia coli. These heterologous products produced by L. casei were detected in the growth medium and parenteral immunization with this medium evoked antibodies against hMBP, confirming that secretion indeed had occurred. Based on the different localization of the heterologous proteins, lactobacilli expressing surface-anchored TTFC are ideally suited for the induction of antibody responses, whereas lactobacilli that secrete myelin proteins can be used for the induction of peripheral T-cell tolerance. In conclusion, the specific technology described here allows the construction of a wide array of safe live recombinant lactobacilli which may prove to be useful in oral intervention strategies for the prevention of infectious diseases or treatment of autoimmune diseases.
Maassen
C B
CB
Division of Immunological and Infectious Diseases, TNO-Prevention and Health (TNO-PG), Leiden, Netherlands.
Laman
J D
JD
den Bak-Glashouwer
M J
MJ
Tielen
F J
FJ
van Holten-Neelen
J C
JC
Hoogteijling
L
L
Antonissen
C
C
Leer
R J
RJ
Pouwels
P H
PH
Boersma
W J
WJ
Shaw
D M
DM
D000284:Administration, Oral; D000818:Animals; D001428:Bacterial Vaccines / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D002417:Cattle; D005434:Flow Cytometry; D005822:Genetic Vectors; D006168:Guinea Pigs; D006801:Humans; D007108:Immune Tolerance; D007150:Immunohistochemistry; D007780:Lacticaseibacillus casei / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D009103:Multiple Sclerosis / Q000276:immunology* / Q000628:therapy; D004676:Myelin Basic Protein / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D010446:Peptide Fragments / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D013742:Tetanus / Q000276:immunology / Q000517:prevention & control*; D013744:Tetanus Toxin / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*
D001428:Bacterial Vaccines; D004676:Myelin Basic Protein; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D013744:Tetanus Toxin; C040635:tetanus toxin fragment C