| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2000: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1999: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Research Abstract |
Antigen-encoding plasmid DNA immunization can induce cellular and humoral immune responses against a variety pathigens, including viruses, parasites, and bacteria, and tumor cells. In previous studies, most of the plasmid DNA was applied either intramuscularly or intradermally and should be taken up by muscle cells or keratinocytes of injection site. The protein produced by plasmid DNA vaccines in the cells are similarly processed and presented by major histocompatibility complex class I and II molecules, resulting in the induction of T helper cells and plasma cells. This process mimics the natural infection by virus.
For the development of vaccines against oral and phayngeal pathogens invading the mucosal epitheria, both secretory and serum immunoglobrin (Ig) A and IgG antibodies and cytotoxic T lymphocytes have been induced. We used a novel approach, targeted muscle immunization, using plasmid pMD701 or pMD702, encoded for Porphyromonas gingivalis outer membrane protein (40-kDa OMP).
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In this study, we obtained good humoral and cell-mediated immune responses in BALB/c mice by muscle administration using the abovementioned DNA vaccine. The production of 40-kDa OMP specific antibody in serum was significantly stimulated by muscle immunization.
1. Injection of DNA vaccine into muscle cell elicited high-level production of antigen-specific IgG antibody. Following the first immunization, 40-kDa OMP antibody was produced in pMD701 or pMD702. Negative control group that was vaccinated with the uniserted pcDNA 3.1 did not exhibit any detectable immune responses to 40-kDa OMP.Specific antibody immune response in serum from DNA-immunized mice showed throughout the experiment from days 0 to 77.
2. P.gingivalis vesicles or S.gordonii cells did not aggregate by themselves. However, aggregation was significant when P.gingivalis vesicles were added to the S.gordonii cells suspension. The aggregation activity of P.gingivalis vesicles toward S.gordonii was partially inhibited by the addition of DNA-immunized mice serum.
These results indicated that intramuscular immunization with plasmid DNA may represent a genetic immunization strategy against infection by oral and pharyngeal pathogens that may invade local, mucosal surface. Less
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