The comparison of the ability of polysaccharide from different Porphyromonas gingivalis in inducing THP-1 cells to produce cytokines / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To compare the ability of the polysaccharide from various Porphyromonas gingivalis (Pg) type and clinical strains in inducing THP-1 cells to produce cytokines interleukin(IL)-1β, IL-8, and tumor necrosis factor(TNF)-α, in order to analyze the immunogenicity of Pg polysaccharide components and the virulence-associated factors of this periodontal pathogen.</p><p><b>METHODS</b>The bacterial polysaccharide was extracted from high virulent Pg strains, W83, SJD2, SJD12 and low virulent Pg, ATCC33277, SJD4, SJD5, and SJD11 by phenol-water extraction. The extracted polysaccharide was used to stimulate the THP-1 cells with different simulation periods and doses. The level of the cytokines, including IL-1β,IL-8 and TNF-α in the cell culture suspension was measured by enzyme-linked immunosorbent assay(ELISA).</p><p><b>RESULTS</b>The polysaccharide extraction of Pg strains was composed of lipopolysaccharide(LPS) and capsular polysaccharide. The secretion of IL-1β, IL-8 and TNF-α, produced by the THP-1 cells showed in a time- and dose-dependent manner in the medium containing 10% fetal bovine serum. The level of these cytokines of the high virulent strains was higher than that of the low virulent strains in medium containing 1% fetal bovine serum.Four hours after stimulation with polysaccharide extracted from high virulent strains, the levels of IL-1β,IL-8, and TNF-α in the cell suspension were (1 639 ± 497), (1 648 ± 513) and (140 ± 48) µg/L, respectively, whereas for low virulent strains, the levels of IL-1β, IL-8, and TNF-α were (773 ± 382), (892 ± 400) and (67 ± 33) µg/L, respectively.</p><p><b>CONCLUSIONS</b>Polysaccharide extracted from Pg could induced the THP-1 cells to secrete the cytokines of IL-1β, IL-8 and TNF-α. The level of the cytokines produced by the THP-1 cells associates with the bacterial virulent properties.</p>

Effects of electronic moxibustion on immune response (II) / 全日本鍼灸学会雑誌

In the previous papers, we reported the effects of electronic moxibustion on immune response of experimental rats to the exogeneous antigens, human γ-globulin.<br>The results supported the theory, “non-specific heat aggregeted autologous tissue protein stimulation therapy” presented by Dr. Shimetaro Hara in 1933.<br>Therefore, in this paper we chose two kinds of antigens, one is the T-cell dependent antigen, dinitrophenylated keyhole limpet hemocyanin (DNP-KLH), the other is the T-cell independent antigen dinitrophenylated Ficoll (DNP-Ficoll) to analyse the mechanism of electronic moxibustion whether it enhances the immune response or not.<br>Using 9 weeks old femal SLC-Wistar rats, we administered the electronic moxibustion according to the method reported in the previous papers. Following daily moxibustion for 8 weeks, antigens were giver twice at intervals of one week together with Freund's complete adjuvant. And 4 days later from the last antigen stimulation direct, DNP plaque forming cells in the spleen were counted.<br>The results obviously showed daily electronic moxibustion for 8 weeks enhanced immune response against the T-cell dependent antigen (DNP-KLH) stimulated rats but no effect on the immune response to the T-cell independent antigen (DNP-Ficoll) stimulated rats.<br>The daily electronic moxibustion for 4 weeks to rats failed to show any effective results against both antigens stimulation.<br>The responses of spleen cells against mitogenic lectins, PHA, Con A and PWM were analysed 3 days after the incubation with lectins by tritiated thymidine up takes into cells. The results also showed the animal group received the electronic moxibustion for 8 weeks manifested higher response against the one of T-cell mitogens, Con A compared with either the group received the electronic moxibustion for 4 weeks or the control group, not received any treatment.<br>These results suggested that the immune activation mechanism exhibited by the electronic moxibustion is via the activation of T-cell function and the electronic moxibustion does not act on B cell nor antibody forming cells.<br>The direct effects on the animal skin by the electronic moxibustion were shown exactly the same physical characteristics as the conventional moxibustion method as reported in the previous papers. Therefore, we could expect the similar T-cell activation effect on the immune response by the conventional moxibustion.<br>But from our results to get such a T-cell activation by the electronic moxibustion, it has been necessary to administrate the electronic moxibustion daily at least for more than 4 weeks.<br>Next we would like to make clear what kinds of subpopulation in the T-cell populations are activated by the electronic moxibustion.<br>Before the clinical administration of the electronic moxibustion as one of immune activators, it is necessary to investigate further about the optimal amounts of the moxibustion, effects of the moxibustion on the cellular immunity or tumor immunity.

Effects of Electronic Moxibustion on Immune Response I / 全日本鍼灸学会雑誌

Great interest has been paid to moxibustion from many thousand years ago as one of effective folk medicine, hewever while through the years only practical use has been emphasized, its scientific basis has remained unclear. About 60 years ago Dr. Shimetaro Hara studied on moxibustion histologically and pharmacologically and suspected the widespread meridian theory (theory of Keiraku) in explaination of the moxibustion effect and presented “non-specific heat aggregated autologous tissue protein therapy” theory. It can be said that his theory coincides with today's nonspecific immune regulatory therapy applied to cancer and immune deficient diseases.<br>Using 9 Week-old femal SLC-Wistar rats, we administered regular moxa moxibustion or electrical moxibustion under the same circumstances as regular moxa moxibustion daily fom definite duration. Following moxibustion, using 0.5mg of HG as an antigen together with Freund's incomplete adjuvant, we sensitized two sites on the foot pads of rats once or twice (2 weeks later).<br>On the 7th day after the primary or secondary sensitization 1.0mg of HγG in 0.1ml of saline was injected subcutaneously at an intact sites of foot pad and foot pad edema formed was measured periodically. Taking sheep red blood cells and using refined human IgG myeloma protein as an antigen and glutar-aldehyde an a fixing reagent, we admindstered PHA (passive hamagglutination) for the assay of serum antibody level of moxibusted animals.<br>In comparison with moxibustion, on the same schedule 5mg/kg of levamisole (LEV) was adminstered orally daily and results were examined.<br>The inflammatory edematous reaction which was induced with the HγG reached a peak 3 hour salter the antigen challenged on the intact foot pad, then gradually weakened until it returned to normal was an immediate type skin reaction.<br>This edema rection in the moxibustion group and the LEV group also when compared with the control group was significantly stregthened. The antibody titer according to the PHA reaction showed after the primary sensitization, no remarkable increase in the moxibustion group, in fact, the level was about the same as the control. After the secondary sensitization the antibody titer of the moxibustion group was much higher than that of the LEV group compared with the control.<br>And the strongest effects were obtained in the moxibustion and LEV group. As an immune activator, the functional mechanism of moxibustion compared with levamisole which is said to have some function on the T cells will become clear in the future.<br>Moreover, it will be clinically possible to use moxibustion as a supplementary therapy to build up the immune response.