Effect of Cytokines produced by 3 genera of periodontopathic bacteria on immune response, and identification of gamma/delta T cell subclass

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The purpose of this experiment was designed to compare the primary and secondary immune responses of mice against periodontal pathogens. We used 3 genera of pathogens : Actinobacillus actinomycetem-comitans. Porphyromonas gingivalis and Fusobacterium nucleatum. For immunization we used each 10 mig of wet weight bacteria injected ip, ip and sc 3 times every week for 3 weeks. Antibody and 7 kinds of cytokine ( TNF-alpha, IL-6, IL-1BETA, IL-lO, IL-4, IL-12 and IFN-gamma) of serum and peritoneal washing after bacterial administration were titrated by ELISA.Results were as follows.Primary response to the same optical concentration of bacteria.1. Comparison of the total cell number after intraperitoneal. administration of thebacteria showed that A.actinomycetemcomitans was the best inducer of exudate cells and P.gingivalis was worst inducer among them.2. In primary antibody response, IgG was dominant A.actinomycetemcomcomitans and P.gingivalis immunization but 1gM was dominant F.nucleatum ad … More ministration.3. Early phase cytokines ie. TNF-alpha, IL-B and IL-1BETA were produced within 24 hours. TNF-a was dominant in serum, IL-6 was in serum and peritoneal washing and IL-1BETA was dominant in peritoneal washings. IL-10 was detectable in the early phase in both serum and the abdomen.4. The cytokines in inductive phase of immunity.IL-10 and IL-4 were detectable in serum only after administration of A.actinomycetemco, itans. IL-12 was detectable but in only in a small amount. IFN-y was only detectable in the serum of mice which had received the bacteria. However in the abdomen IFN- was found in high level nly the mice treated with F.nucleatum.5. The response of immunized mice after administration of bacteria.A high level of IFN-r was produced on day 1 both in serum and the peritoneal washings.6.gamma/delta T cell subset induced by F.nucleatum injection.The gamma/delta TCR subset induced by F.nucleatum injection during the primary responce was determined as Vgamma_6 and Vdelta_1 by the RTCR method using specific primer of gamma and delta. Less