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Biomedical subjects

H Shimauchi

Publications and source records attributed to H Shimauchi.

At least 37 records · Page 2Linked to original sources
PubMed1996

Development of a quantitative sampling method for periapical exudates from human root canals.

A quantitative method for collection of periapical exudates on endodontic paper points is described. In a pilot study, it was revealed that the relationship between fluid volume and wetted length of paper points had a highly significant curvilinear relationship (p < 0.0001), and a highly significant positive linear relationship was found to describe eluted and absorbed interleukin (IL)-1 beta activities from paper points. Periapical exudates from 29 root canals with apical periodontitis were collected using this method, and IL-1 beta activities in clinical specimens were measured. Periapical exudates (0.15 to 26.7 microliters) were recovered, and the range of IL-1 beta concentration was 0.1 to 179.5 ng/ml. These results showed that this sampling method was useful to analyze immunological changes in periapical lesions.

Dental Pulp Cavity↗
PubMed1996

Relationship between prostaglandin E2 concentrations in periapical exudates from root canals and clinical findings of periapical periodontitis.

In this study the relationship of prostaglandin E2 (PGE2) concentrations in periapical exudates with clinical findings of teeth with periapical periodontitis is discussed. Periapical exudate samples were obtained from root canals of 77 endodontically involved teeth during routine root canal treatment by the quantitative sampling method using paper points. PGE2 concentrations in periapical exudates (PE-PGE2) were determined by radioimmunoassay. Significantly higher levels of PGE2 were found in periapical exudates from teeth with radiolucent areas than from teeth without radiolucent areas (236.8 +/- 521.3 pg/microliter vs. 14.9 +/- 23.1 pg/microliter, respectively). The elevated PE-PGE2 levels were associated with the presence of clinical symptoms that reflected an acute inflammation in the periapical lesion. In contrast, a significantly negative association of decreased PE-PGE2 levels with increasing size of radiolucent areas was demonstrated. These results suggested that PGE2 was produced locally in periapical lesions and that the PGE2 concentration in periapical exudate could reflect the state of the disease activity in periapical periodontitis.

Chi-Square Distribution↗
PubMed1996

Stimulation of splenocytes in C3H/HeJ mice with Porphyromonas gingivalis lipid A in comparison with enterobacterial lipid A.

Porphyromonas gingivalis 381 lipid A induced strong mitogenic response in splenic B cells separated from LPS-non-responsive C3H/HeJ mice as well as LPS-responsive C3H/HeN mice, by using a magnetic cell sorting system. The lipid A also exhibited mitogenic activity in splenic T cells. P. gingivalis lipid A induced lower production of interleukin-1 beta (IL-1 beta) in splenic macrophage cultures and exhibited a comparable IL-6 producing activity in splenic B cells of C3H/HeN mice as compared to Escherichia coli-type synthetic lipid A (compound 506) and monophosphoryl lipid A from Salmonella minnesota Re 595 (MLA). Furthermore, P. gingivalis lipid A, but not compound 506 nor MLA, induced low IL-1 beta and high IL-6 production in C3H/HeJ mice. C3H/HeJ T cells in response to P. gingivalis lipid A stimulation resulted in definite IL-2 mRNA and its production, whereas IL-4 response was scarcely active in both C3H/HeN and C3H/HeJ T cells. P. gingivalis lipid A increased tyrosine, serine/threonine phosphorylation of proteins with various major induced bands in splenic B cells of both C3H/HeN and C3H/HeJ mice. Thus, P. gingivalis lipid A stimulated the splenic immunocytes of C3H/HeJ as well as C3H/HeN mice.

Adjuvants, Immunologic↗
PubMed1996

Diagnostic strategies of periodontitis based on the molecular mechanisms of periodontal tissue destruction.

OBJECTIVE: Periodontitis is a disease showing differences in disease progression between patients and between sites within a patient. Routine clinical examinations today are not useful enough to distinguish susceptible patients and active lesions from resistant patients and chronic lesions. Diagnostic markers should be pathogenic and inflammatory factors participating in periodontal tissue destruction. These are both local and systemic factors. MATERIALS AND METHODS: First of all, pathogenic factors and proinflammatory cytokines or mediators in gingival crevicular fluid (GCF) were examined and the difference was found between active and inactive periodontitis lesions distinguished by attachment loss. Active lesions were detected by discriminant-function analysis of these examinations, although the sensitivity of differential diagnosis was low. Then, we established a novel needle biopsy for understanding the pathophysiological conditions elicited in active and chronic inflammatory processes of periodontal tissue destruction. A variety of cytokines and mediators were detected in biopsied specimens by reversed transcription polymerase chain reactions (RT-PCR). Cytokine profiles were varied in inflammed periodontal biopsies. As IFN gamma mRNA expression was enhanced in inflamed gingiva, antigen-presenting-cell (APC) functions of human gingival fibroblasts (HGF) were examined. RESULTS: Despite the phenotypical resemblance of IFN gamma-treated HGF to so-called APC, HLA-DR positive HGF could not induce proliferation but suppressed proliferation of alloreactive peripheral blood T cells (PBT). However, HLA-DR positive HGF stimulated the proliferative responses of PBT which had been primed with allo-APC. Regulatory immune responses by IFN gamma were different in T cell conditions. CONCLUSIONS: Various kinds of cytokines participated in periodontal inflammation, and every cytokine is multi-functional. Complex and compound inflammatory processes can be clarified by examining cytokine networks and the precise effects of each cytokine on each of the cell types comprising periodontal tissue. It is, therefore, necessary for establishing diagnostic strategies to integrate pathogenic and inflammatory factors in periodontal tissue destruction.

Antibodies, Bacterial↗
PubMed1995

Oral immunoadjuvant activity of lipophilic derivatives of N-acetylglucosaminyl-beta(1-->4)-N-acetylmuramyl-L-alanyl-D-isoglutamin yl-(L)- 2,6-meso-diaminopimeric acid-(D)-amide.

Twenty-four kinds of the acylated, amidated or esterified derivatives of N-acetylglucosaminyl (GlcNAc)-beta(1-->4)-N-acetylmuramyl (MurNAc)-L-alanyl (Ala)-D-isoglutaminyl (isoGln)-(L)-meso-2,6-diaminopimeric acid (A2pm)-(D)-amide (GMP3-A) which were prepared by chemical modifications of an enzymatic hydrolysate of Lactobacillus plantarum cell-wall peptidoglycans were examined for oral adjuvant activity by gastric intubation with bovine serum albumin (BSA) in liposomes into BALB/c mice. The gastric intubation of GlcNAc-beta(1-->4)-MurNAc-L-Ala-D-isoGln-(L)- stearoyl-(D)-meso-A2pm-(D)-amide-(L)-O-octyl, -nonyl and -dodecyl esters exhibited the most marked oral adjuvant activity in terms of enhanced production of serum anti-BSA IgG antibody. Some derivatives showed a less marked adjuvanticity and others were totally inactive. Thus the oral adjuvanticity of test lipophilic derivatives of GMP3-A in liposomes was dependent upon their chemical structure.

Adjuvants, Immunologic↗
PubMed1992

Induction of mucosal and serum immune responses to a specific antigen of periodontal bacteria.

The dynamics of the host immune response to periodontal bacteria not only may be informative from the standpoint of specific mucosal protection to these pathogens, but also may reveal the capacity of the mucosal immune response to provide protection of the host. To this end, we have examined the immune response to chromatographically purified fimbriae of P. gingivalis administered orally or systemically with liposomes and adjuvant in BALB/c mice, high responders to this antigen. Oral administration of P. gingivalis fimbriae clearly enhanced the fimbriae-specific salivary IgA response. ELISPOT analysis revealed that significant numbers of fimbriae-specific IgA SFC were seen in lamina propria and mesenteric lymph nodes but not in Peyer's patches of mice immunized orally. In contrast, antigen-specific IgM and IgG SFC were seen mainly in the circulating blood mononuclear cells. On the other hand, subcutaneous injection of fimbriae with GM-53 also raised the fimbriae-specific IgG followed by IgM and IgA responses in serum, and both IgA and IgG responses in saliva. Oral immunization was less effective than subcutaneous injection in terms of the serum antibody response. However, the salivary antibody level of mice injected subcutaneously was similar to that of mice immunized orally. In the subcutaneously immunized mice, fimbriae-specific SFC were detected in the spleen, blood, and brachial lymph nodes by ELISPOT assay. Fimbriae-specific IgM SFC appeared earlier and antigen-specific IgG SFC were seen later. These results show that the combined use of fimbriae together with the adjuvant results in sharply increased IgA responses in saliva and IgG responses in serum. In summary, it is clear that the nature of the host's antibody response in serum and mucosal secretions is distinct, and depends on the route of antigen administration, the use of adjuvant and/or liposomes, and the temporal phase of the humoral immune response following various immunization regimes.

Animals↗
PubMed1992

Immunochemical and biological characterization of outer membrane proteins of Porphyromonas endodontalis.

Outer membrane proteins (OMP) of Porphyromonas endodontalis HG 370 (ATCC 35406) were prepared from the cell envelope fraction of the organisms. The cell envelope that had been obtained by sonication of the whole cells was extracted in 2% lithium dodecyl sulfate and then successively chromatographed with Sephacryl S-200 HR and DEAE-Sepharose Fast Flow. Two OMP fractions, OMP-I and OMP-II, were obtained, and their immunochemical properties and induction of specific antibodies were examined. The OMP-I preparation consisted of a major protein with an apparent molecular mass of 31 kDa and other moderate to minor proteins of 40.3, 51.4, 67, and 71.6 kDa, while the OMP-II preparation contained 14-, 15.5-, 27-, and 44-kDa proteins as revealed by sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis. OMP-I was found to form hydrophilic diffusion pores by incorporation into artificial liposomes composed of egg yolk phosphatidylcholine and dicetylphosphate, indicating that OMP-I exhibited significant porin activity. However, the liposomes containing heat-denatured OMP-I were scarcely active. Spontaneous and antigen-specific immunoglobulin M (IgM)-, IgG-, and IgA-secreting spot-forming cells (SFC) enzymatically dissociated into single-cell suspensions from chronically inflamed periapical tissues and were enumerated by enzyme-linked immunospot assay. In patients with radicular cysts or dental granulomas, the major isotype of spontaneous SFC was IgG. In radicular cysts, the OMP-II-specific IgG SFC represented 0.13% of the total IgG SFC, while the antigen-specific IgA or IgM SFC was not observed. It was also found that none of these mononuclear cells produced antibodies specific for OMP-I or lipopolysaccharide of P. endodontalis.

Antibodies, Bacterial↗
PubMed1991

Distribution and immunochemical specificities of fimbriae of Porphyromonas gingivalis and related bacterial species.

Rabbit polyclonal antibody (Poly-1) and mouse monoclonal antibodies (mAbs) TO-11, TO-14 and TO-M1 specific for Porphyromonas gingivalis 381 fimbriae were prepared. Poly-1 and the 3 mAbs were screened for their reactivity with whole cells oral and nonoral black-pigmented bacterial species by enzyme-linked immunosorbent assay (ELISA) and the binding experiment using [125I]Poly-1 and [125I]mAbs. ELISA revealed that Poly-1 definitely reacted with whole cells of all the 11 strains of P. gingivalis tested. However, 8 of 11 P. gingivalis strains reacted with mAbs TO-11, TO-14 and TO-M1. These results were confirmed by the specific binding of radiolabelled Poly-1 and mAb TO-11 to the 8 strains. The M(r) of the fimbrial subunit protein (fimbrilin) isolated and purified from P. gingivalis strains 381, BH18/10, HW24D-1, 6/26 and OMZ 314 was 41 kDa by sodium dodecylsulfate-polyacrylamide gel electrophoresis. It was found by immunoblotting that mAbs TO-11 and TO-14/TO-M1 recognized different epitopes of fimbrial protein from P. gingivalis strains. Immunoelectron micrographs of whole cells and the purified fimbriae of P. gingivalis strains visualized similar serotype-specific antibody bindings to the fimbriae. These results indicate that 11 strains of P. gingivalis could be divided into at least 2 separate groups based on the immunochemical specificities of the fimbriae.

Animals↗
PubMed1991

Immune response gene regulation of the humoral immune response to Porphyromonas gingivalis fimbriae in mice.

Among various strains of mice immunized orally with Porphyromonas gingivalis fimbriae and adjuvant GM-53 in liposomes, BALB/c and DBA/2 mice (H-2d) were found to be high responders to the fimbriae, CBA/J and C3H mice (H-2k) were intermediate, while C57BL/6 mice were low responders in terms of serum IgG and salivary IgA responses. Furthermore, humoral immune responses were examined using congeneic mice of B10 background showing different H-2 haplotypes, and it was revealed that B10.D2 mice (H-2d), followed by B10.BR (H-2k), responded well to antigenic stimulation of the fimbriae, while C57BL/10 mice (B10, H-2b) were low responders to the fimbriae. Hybrids between BALB/c and C57BL/6 mice were found to reflect a phenotype of low responders. Thus, the humoral immune responses to P. gingivalis in mice are restricted by H-2 haplotype.

Animals↗
PubMed1990

Antimicrobial activities of thiolactomycin against gram-negative anaerobes associated with periodontal disease. f1.

Thiolactomycin (TLM), (4R)-(2E,5E)-2,4,6-trimethyl-3-hydroxy-2,5, 7-octatriene-4-thiolide, purified from a culture filtrate of a strain of the Nocardia species, was examined for antimicrobial activities against more than 100 strains of oral and periodontally associated bacteria. Nine other commonly used antibiotics were also included for the test. We found that TLM exhibited strong and selective antimicrobial activities against Bacteroides gingivalis and other oral black-pigmented Bacteroides species that may be etiologically associated with adult periodontitis. TLM also inhibited the growth of Actinobacillus actinomycetemcomitans, but did not affect the growth of oral streptococcal species and Eubacterium species. Strains of Eikenella corrodens were moderately susceptible to TLM, while Actinomyces viscosus strains were only slightly susceptible to it. Other antibiotics used for comparison showed a broad spectrum of antimicrobial activities in general. In conclusion, TLM exhibited highly selective antimicrobial activities to black-pigmented Bacteroides species and A. actinomycetemcomitans, both of which are implicated in the pathogenesis of human periodontal disease.

Actinobacillus↗
PubMed1990

Humoral immune response to Bacteroides gingivalis fimbrial antigen in mice.

Bacteroides gingivalis fimbrial antigen incorporated into liposomes, but not in Tris-HCl buffer, significantly raised the levels of anti-fimbriae antibodies in serum, particularly of the IgG class, after oral primary and booster immunizations in BALB/c mice. An approximately linear relationship was observed between the dose of fimbrial antigen and the level of fimbriae-specific antibodies produced; antibody production reached its maximum at an immunization dosage of 500 micrograms of fimbriae per mouse. Fimbriae-specific antibody production was enhanced by use of a semi-synthetic adjuvant, a stearoyl derivative of sodium beta-N-acetylglucosaminyl-(1----4)-N-acetylmuramyl-L-alanyl-D-isoglutaminyl-(L) - stearoyl-(D)-meso-diamino-pimelic acid-(D)-amide-D-alanine (GM)-53) in liposomes. High anti-fimbriae antibody levels in serum and saliva were maintained for several months in the mice that had received two orally administered boosters of fimbrial antigen with GM-53 in liposomes. Salivary anti-fimbriae antibody levels, particularly of the IgA class, were markedly raised.

Administration, Oral↗
PubMed1990

[Studies on the induction of the humoral immune responses to Bacteroides gingivalis fimbrial antigen in mice].

Serum and salivary antibody responses to Bacteroides gingivalis fimbriae administered either orally or subcutaneously (s.c.) with or without an adjuvant in various strains of mice were examined in this study. Following results were obtained. 1) Oral administration of B. gingivalis fimbriae with GM-53 as an adjuvant in liposomes, but not in Tris-HCl buffer, definitely enhanced the fimbriae-specific IgG responses, mainly IgG1 followed by IgG2b, IgG2a and IgG3 in serum and IgA response in saliva of BALB/c mice. On the other hand, s.c. injection of fimbriae with GM-53 or MDP-Lys (L18) also raised the fimbriae-specific IgG followed by IgA and IgM responses in serum, and both IgA and IgG responses in saliva of BALB/c mice. Oral immunization was less effective than s.c. injection in terms of the production of serum antibody in the mice. However, the level of salivary antibody of mice injected s.c. was similar to that of mice immunized orally. 2) High anti-fimbriae antibodies in serum were maintained in BALB/c mice immunized orally with fimbriae and GM-53 in liposomes for approximately 7 months after the primary immunizations. Oral administration also induced and held the fimbriae-specific IgA response in saliva for at least 6 months after the primary immunizations. The levels of fimbriae-specific IgA in saliva after the second boosters on days 123 and 124 were higher than those after the primary ones on days 27 and 28. 3) Among various strains of mice immunized orally with fimbriae and GM-53 in liposomes, BALB/c and DBA/2 mice (H-2d) significantly produced high levels of both serum IgG and salivary IgA antibodies specific for fimbriae. Furthermore, B10.D2 mice (H-2d) were responders followed by B10.BR (H-2k), while C57BL/10 mice (B10, H-2b) were low responders to the fimbriae. These results show that the combined use of fimbriae together with an adjuvant results in a sharply increased IgA antibody response in saliva and a predominantly stimulated IgG antibody in serum, and it was suggested that these responses are restricted by H-2 haplotype.

Acetylmuramyl-Alanyl-Isoglutamine↗
PubMed1989

Mucosal and systemic immune responses in BALB/c mice to Bacteroides gingivalis fimbriae administered orally.

A 41,000-molecular-weight fimbrial protein was isolated from freshly cultivated whole cells of Bacteroides gingivalis 381 and purified chromatographically. Salivary and serum antibody responses to the fimbriae, which had been orally administered in the presence of an acyl derivative of muramylpeptides, i.e., either N2-[(N-acetylmuramyl)-L-alanyl-D-isoglutaminyl]-N6-stearoyl-L-lysine [MDP-Lys(L18)] or sodium beta-N-acetyl-glucosaminyl-(1----4)-N-acetylmuramyl-L-alanyl-D-isoglu tam inyl- (L)-stearoyl-(D)-meso-2,6-diaminopimelic acid-(D)-amine-D-alanine (GM-53), or in the absence of adjuvant, were examined in BALB/c mice when administered by gastric intubation on days 0 and 1 as primary immunizations and on days 27 and 28 as booster immunizations. Gastric intubation of the fimbriae with an adjuvant significantly enhanced the production of anti-fimbria immunoglobulin A (IgA) in saliva. Subcutaneous injection of fimbriae along with an adjuvant also raised anti-fimbria IgA levels, as well as IgG levels, in saliva. Both immunization procedures enhanced the levels of anti-fimbria IgG, IgA, and IgM in serum, and the major class of fimbria-specific antibody was IgG, followed by IgA and IgM. However, subcutaneous injection was more effective than gastric intubation to enhance the production of serum antibody in mice. The subclasses of IgG antibody specific for fimbriae in serum were mainly IgG1, followed by IgG2a, IgG2b, and IgG3. These results demonstrated that the combined use of B. gingivalis fimbrial antigen and either GM-53 or MDP-Lys(L18) resulted in a sharply increased IgA antibody response in saliva and a predominantly stimulated IgG antibody response in serum, respectively. Both antibodies were found to be specific for the fimbriae used for immunization.

Adjuvants, Immunologic↗
PubMed1987

Possible existence of a novel amphipathic immunostimulator in the phenol-water extracts of Mycobacteriaceae.

The extracts having diverse immunostimulating activities were obtained as a water-phase fraction from four bacterial species representing the 4 genera (Mycobacterium, Nocardia, Gordona, and Rhodococcus) of Mycobacteriaceae by the phenol-water method, which is commonly used for extraction of endotoxic lipopolysaccharides (LPS) from gram-negative bacteria and amphipathic substances from gram-positives. These fractions, especially those of G. aurantiaca and R. terrae, showed strong stimulatory effects on murine splenocytes, macrophages of mice and guinea pigs, the immunoadjuvant activities in guinea pigs and mice, and the distinct activities inducing a tumor necrosis factor and interferons alpha/beta and gamma in primed mice. The fractions from G. aurantiaca and R. terrae exhibited potent pyrogenicity and the ability to activate the clotting enzyme cascade of the horseshoe crab (Tachypleus tridentatus). Some of these biological activities were not very different from the potency of the reference endotoxic LPS derived from Escherichia coli or Fusobacterium nucleatum. But the test fractions neither showed the activity to prepare rabbit skin to the local Shwartzman reaction, nor reacted with anti-lipid A conventional and monoclonal antibodies. Furthermore, unlike LPS, these fractions stimulated the splenocytes of C3H/HeJ mice (LPS-Nonresponder). Although the fractions showing the above biological activities have not yet been adequately purified, they contained polysaccharides, whose main constituent sugar is mannose with a smaller amount of arabinose, fatty acids consisting primarily of palmitic, stearic, and tuberculostearic acids, and small amounts of peptides and amino sugars. Since components characteristic of known immunomodulators of bacterial origin, namely endotoxins (lipid A's), cell wall peptidoglycans, lipoteichoic acids, cord factors (trehalose dimycolates), or deoxyribonucleic acids, were practically not detected in these fractions, the agent responsible for the above bioactivities is considered to be a novel substance different from the known, bacterial immunomodulators.

Adjuvants, Immunologic↗
PubMed1986

Immunobiological activities of synthetic lipid A analogs with low endotoxicity.

Synthetic lipid A analogs, beta(1-6)glucosamine disaccharide 1,4'-bisphosphates, which possesses four tetradecanoyl groups at the 2- and 2'-amino, and 3- and 3'-hydroxyl groups (LA-17-PP), and each two of the (R)-3-hydroxytetradecanoyl and tetradecanoyl groups at the 2- and 2'-amino and 3- and 3'-hydroxyl groups, respectively (LA-18-PP), were far less endotoxic than synthetic (506, LA-15-PP) and bacterial Escherichia coli type lipid A's; neither compound showed any detectable lethal toxicity in chicken embryos or preparatory activity for the local Shwartzman reaction in rabbits. Also both compounds were only weakly pyrogenic and comparably less lethally toxic in galactosamine-loaded mice than the reference synthetic and bacterial lipid A's and a synthetic counterpart to biosynthetic lipid A precursor Ia (406, LA-14-PP). Nevertheless, LA-17-PP and LA-18-PP exhibited definite in vivo immunoadjuvant activity in mice, and the ability to induce a possible tumor necrosis factor and alpha/beta interferon in Mycobacterium bovis BCG and Propionibacterium acnes-primed mice, respectively, although these activities were weaker than those of the reference lipid A's. 4'-Monophosphate analogs of the above two test compounds exhibited neither endotoxic nor beneficial activities, but they showed remarkable in vitro bioactivities comparable to those of the corresponding bisphosphate compounds; the ability to activate the human complement system and the clotting enzyme cascade of horseshoe crab amoebocyte lysate, stimulatory effects on guinea pig and murine peritoneal macrophages, and murine splenocytes.

Adjuvants, Immunologic↗