title: Protection against Mycoplasma pulmonis infection by genetic vaccination.
abstract: The induction of an immune response against a foreign protein usually requires purification of that protein, which is injected into animals. The isolation of pure protein is time consuming and costly. Recently, a technique called biolistic transformation (biological ballistic system) microparticle injection, gene gun, or particle bombardment was developed. The basic idea is that DNA or biological material coated onto heavy tungsten or gold particles is shot into target cells or animals. We have vaccinated mice by introducing the gene (Mycoplasma pulmonis DNA or a specific fragment) encoding a protein recognized by a protective monoclonal antibody directly into the skin or muscle of mice by two methods: (i) using a hand-held form of the biolistic system that can propel DNA-coated gold microprojectiles (2 micrograms of DNA) directly into the skin; (ii) using a conventional intramuscular injection of the DNA (100 micrograms) into quadricep muscles of transfected mice. HeLa cells were transfected in vitro by the gene gun or by the liposomal delivery system. Indirect immuno-fluorescent antibody (IFA) assay of culture cells indicated that both methods could be successful. Production of antibody and cell-mediated immunity against M.pulmonis were monitored by assaying serum IFA and enzyme-linked immunosorbent assay (ELISA), and delayed type hypersensitivity. In addition, macrophage migration inhibition and lymphocyte transformation to antigen in spleen cells were also tested. Both delivery systems induced humoral and cellular immunity, and vaccinated the mice against infection. Genetic immunization by using the gene gun saves time, money, and labor; moreover, this general method is also applicable to gene therapy.
authors.affiliation: Department of Pathology, University of Texas Southwestern Medical Center at Dallas 75235, USA.
authors.forename: W C
authors.initials: WC
authors.lastname: Lai
authors.forename: M
authors.initials: M
authors.lastname: Bennett
authors.forename: S A
authors.initials: SA
authors.lastname: Johnston
authors.forename: M A
authors.initials: MA
authors.lastname: Barry
authors.forename: S P
authors.initials: SP
authors.lastname: Pakes
mesh_terms: D000818:Animals; D000917:Antibody Formation; D000942:Antigens, Bacterial / Q000032:analysis / Q000096:biosynthesis; D001428:Bacterial Vaccines; D001483:Base Sequence; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics / Q000276:immunology*; D005260:Female; D005798:Genes, Bacterial; D015698:Genomic Library; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009174:Mycoplasma / Q000235:genetics*; D009175:Mycoplasma Infections / Q000276:immunology* / Q000517:prevention & control; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis
chemical_list: D000942:Antigens, Bacterial; D001428:Bacterial Vaccines; D017931:DNA Primers; D004269:DNA, Bacterial; D011994:Recombinant Proteins