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

本实验的目的是比较小鼠对牙周病原体的原发和继发免疫反应。我们使用了3个属的病原菌：伴放线放线杆菌。牙龈卟啉单胞菌和核梭杆菌。免疫采用每10毫克湿重细菌ip、ip和sc，每周3次，共3周。用ELISA法检测细菌给药后血清和腹腔液中的抗体和7种细胞因子(肿瘤坏死因子-α、IL-6、IL-1β、IL-10、IL-4、IL-12和干扰素-γ)。腹膜腔内注射后细胞总数的比较。给药结果表明，伴生放线菌是渗出细胞的最佳诱导剂，牙龈假单胞菌是最差的诱导剂。在一次抗体应答中，伴生放线放线菌和牙龈假单胞菌免疫以Ig G为主，核杆菌和…免疫以1gm为主。更多的服务。早期细胞因子，即。24小时内产生肿瘤坏死因子-α、白介素B和白介素1β。血清以肿瘤坏死因子-α为主，血清和腹膜冲洗液以IL-6为主，腹膜冲洗液以IL-1β为主。IL-10在早期血清和腹部均可检测到。免疫诱导期的细胞因子IL-10和IL-4只有在接种放线菌后才能在血清中检测到。IL-12可检测到，但仅有少量。干扰素-γ只在接受细菌感染的小鼠的血清中检测到。而在小鼠的腹部，只有F-核酸菌处理的小鼠体内有较高水平的干扰素。免疫小鼠接种细菌后的应答。第1天血清和腹膜冲洗液中均产生高水平的干扰素-r。6.核杆菌免疫小鼠诱导的γ/Delta T细胞亚群。用γ和Delta的特异性引物，用RTCR方法测定核杆菌免疫小鼠在初次应答过程中诱导的T细胞亚群为VGamma_6和VDelta_1。较少

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