title: Oral vaccination with Lactococcus lactis expressing the Vibrio cholerae Wzm protein to enhance mucosal and systemic immunity.
abstract: A gene associated with lipopolysaccharide (LPS) transport was cloned from a local clinical Vibrio cholerae O1 strain of the Ogawa serotype by using the Lactococcus lactis nisin-controlled expression (NICE) system. The V. cholerae wzm gene, which codes for an integral membrane transporter protein, was expressed and targeted to the cytoplasmic membrane, and was crudely isolated through simple centrifugation and SDS solubilization. To examine seroreactivity of this construct, rabbits were orally fed with 10(9) cfu/ml of live, recombinant L. lactis carrying the wzm gene, induced with nisin prior to administration. Recombinant plasmids were retrieved from L. lactis cultured directly from stool samples of inoculated rabbits. Reverse-transcriptase PCR of wzm using the retrieved plasmids confirmed transcription of this gene, indicating viability and stability of the recombinants in vivo. The L. lactis-Wzm construct elicited substantial levels of IgG and sIgA, and challenge with virulent V. cholerae O1 evoked severe diarrhoea in the naive, non-immunised control group, but not in those fed with either recombinant or non-recombinant L. lactis. Oral administration with recombinant L. lactis expressing the V. cholerae wzm gene increases both systemic and mucosal immunity, whereas L. lactis itself appears capable of protecting against the diarrhoeal symptoms caused by V. cholerae. Wzm is a conserved membrane protein associated with the LPS endotoxin, and together with the food-grade L. lactis, represent an attractive target for the development of a safer, live anti-infective therapy against V. cholerae.
authors.affiliation: Medical Microbiology and Parasitology Laboratory, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia. hana_zamri@yahoo.com
authors.forename: Hana Farizah
authors.initials: HF
authors.lastname: Zamri
authors.forename: Mariana Nor
authors.initials: MN
authors.lastname: Shamsudin
authors.forename: Raha Abdul
authors.initials: RA
authors.lastname: Rahim
authors.forename: Vasanthakumari
authors.initials: V
authors.lastname: Neela
mesh_terms: D000284:Administration, Oral; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology*; D002771:Cholera / Q000276:immunology / Q000517:prevention & control*; D022121:Cholera Vaccines / Q000008:administration & dosage / Q000276:immunology*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D003967:Diarrhea / Q000276:immunology / Q000517:prevention & control; D004195:Disease Models, Animal; D005822:Genetic Vectors; D007071:Immunoglobulin A, Secretory / Q000097:blood; D007074:Immunoglobulin G / Q000097:blood; D013294:Lactococcus lactis / Q000235:genetics*; D008297:Male; D026901:Membrane Transport Proteins / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D010957:Plasmids; D011817:Rabbits; D017422:Sequence Analysis, DNA; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000276:immunology; D041241:Vibrio cholerae O1 / Q000235:genetics / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D022121:Cholera Vaccines; D004269:DNA, Bacterial; D007071:Immunoglobulin A, Secretory; D007074:Immunoglobulin G; D026901:Membrane Transport Proteins; D014614:Vaccines, Synthetic