Protection AgainstMycoplasma pulmonisInfection by Genetic Vaccination

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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 a 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 the 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 μg of DNA) directly into the skin; (ii) using a conventional intramuscular injection of the DNA (100 μg) into quadricep muscles of transfected mice. HeLa cells were transfected in vitro by the gene gun or by the liposomal delivery system. Indirect immunofluorescent 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.