Current developments in molecular biology and genetics have strongly influenced the traditional concept of inflammatory disorders. One example, rheumatoid arthritis (RA) has been well recognized as a chronic“inflammatory”disorders unknown etiology. However, we have revealed a role one of oncogenic retrovirus HTLV-I tax gene in protooncogene expression in synovial tissue and these events may be associate with synovial hyperplasia RA. One of the pathological features of SCs in active RA is the unusual proliferative of heterogeneous cells termed rheumatoid synoviocytes. Several reports suggest that the nature of rheumatoid synoviocytes is similar to tumor-like expansion. Following the HTLV-Itaxgene association, we first established immortalized synoviocytes clones bearing oncogenic properties from heterogeneous synoviocytes. Clonal and molecular biological analysis clearly suggest that rheumatoid arthritis seems to be one of the characteristics with genetic aberrancy.
These studies suggested that chronic inflammatory disorders should come under new consideration based upon molecular biology or genetics.

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Human neutrophils maximally stimulated with the optimal concentration (100 ng/ml) of phorbol myri-state acetate (PMA), a direct activator of protein kinase C (PKC), for 5 min at 37°C did not respond with superoxide (O^-₂) release to the later addition of PMA itself or the Ca²⁺ionophore ionomycin. The loss of responsiveness to the later addition of ionomycin was dependent on the interval between the addition of PMA and ionomycin. However, these cells did respond with enhanced release of O^-₂to the later addition of N-formyl-methionyl-leucyl-phenylalanine (FMLP) or concanavalin A (Con A) ·In these PMA-pretreated cells, an increase in cytoplasmic free Ca²⁺ ( [Ca²⁺] i) induced by ionomycin was unaffected, whereas that induced by FMLP was inhibited by 50-60% and that induced by Con A was completely abolished. UCN-01, a PKC inhibitor, inhibited O^-₂release induced by PMA, but not by FMLP.
These findings suggest that FMLP and Con A trigger the respiratory burst in human neutrophils by activating the definite pathway which include other signals than activation of PKC and an increase in [Ca²⁺] i.

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We have examined the mechanism of peptide leukotriene (LT) production in rat liver. LTC₄ synthesizing activity was shown to be localized in hepatocyte rather than non-parenchymal cells including Kupffer cell. Stimulation of the isolated hepatocyte with calcium ionophore (A 23187) did not cause any production of peptide LTs, whereas addition of LTA₄induced a remarkable production of peptide LTs without the A 23187 stimulation. Kupffer cell which was shown to generate only a slight amount of peptide LTs with the A 23187 stimulation even in the presence of LTA₄, produced remarkably peptide LTs when LTC₄synthase partially purified from the hepatocyte was included into the culture system. Coculture of hepatocyte markedly enhanced production of peptide LTs.
These results indicate that an enzymatic cooperation between hepatocyte and Kupffer cell would play an important role in generating peptide LTs in rat liver.

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Tumor necrosis factor-α (TNF-α), a cytokine secreted by macrophages, was recently recognized as an important molecule in host defense mechanism. However, its overproduction is also implied in many acute and chronic disease states. The inhibition of TNF-α overproduction, therefore, leads to suppression of acute and chronic inflammation. This work was undertaken to ascertain whether vegetable juice from perilla leaf, which may be a new candidate for an antiinflammatory reagent, can inhibit the endogenous production of TNF-α in mice.
Both red and green perilla juice significantly inhibited TNF-α production byperos administration. Perilla juice showed nearly the same level of an inhibitory activity against TNF-α production as did Saibokuto, an antiinflammatory reagent. The inhibitory activity was stable for heat treatment (100°C, 10 min) . Perilla contains an active molecule (s) which can inhibit a host from releasing TNF-α. This molecule thus may suppress host defense systems as well as antiinflammatory drugs such as glucocorticoid or Saibokuto.

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We evaluated the effect of low-dose long-term erythromycin (EM) therapy (600 mg/day) on activated lymphocytes in the peripheral blood of the patient with Sjögren's syndrome accompanied with lung involvement. Before and after EM therapy, we analysed the subsets of peripheral lymphocytes using two-colour analysis by FACScan. Used monoclonal antibodies were Leu 2a, Leu 3a, Leu 4 and Leu HLA-DR. After 9 months of EM therapy, the percentages of activated Leu 2a positive cells, Leu 3a positive cells, and Leu 4 positive cells significantly decreased.
These results might be useful to clarify the mechanism of low-dose long-term EM therapy against diffuse panbronchiolitis.

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[in Japanese]

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It has recently become apparent that peritoneal fluid and its cellular and humoral constituents are associated with the pathophysiology of pelvic endo metriosis. We examined the expression of cytokine genes in peritoneal mononuclear cells obtained from 53 patients with and without pelvic endometriosis by using the reverse transcription-polymerase chain reaction. We also analyzed the levels of interleukin (IL) -1β and IL-1 receptor antagonist (IL-1ra) messenger RNAs (mRNAs) on Northern blots, and deter mined the level of IL-1β in peritoneal fluids.
The peritoneal mononuclear cells expressed frequently the mRNAs for IL-1β, IL-1ra and tumor necrosis factor (TNF) . There was no difference between the patients with endometriosis and those without endometriosis in the expression rate of all the genes analyzed. The level of IL-1β mRNA expression in peritoneal macrophages was elevated in the stage I pelvic endometriosis group, whereas the increased expression of IL-1ra mRNA was observed in the stages 1ff and N pelvic endometriosis group. The level of IL-1β in peritoneal fluid was elevated in the stage I pelvic endometriosis group, and stage II pelvic endometriosis group.
Our results indicate that peritoneal mononuclear cells express mainly monokine genes such as IL-1β, IL-1ra and TNF, and that peritoneal macrophages express IL-1ra mRNA rather than IL-1β mRNA with the progress of pelvic endometriosis. IL-1 in peritoneal fluid may be responsible for the adhesions and infertility associated with pelvic endometriosis.

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The goal of periodontal therapy is to regenerate the periodontium lost by the progression of periodontitis. In this regeneration process, migration, proliferation, and differentiation of periodontal ligament (PDL) cells are essential. The periodontal ligament tissues are composed of cells and their related extracellular matrix (ECM) components. We examined the effects of extracellular matrix components on biological activities of PDL cells. PDL cells were seeded on the plates coated with type I, III, IV collagen, fibronectin, or laminin. Proliferation of PDL cells was enhanced by fibronectin and laminin. Type I, III, IVcollagen did not enhance the proliferation of PDL cells. ALPase activity of PDL cells was enhanced by type I, III, IV collagen. And type I collagen was the most potent activator. Laminin inhibited the ALPase activity. The formation of calcified nodules by PDL cells was examined by the Alizarin staining methods. Type I collagen promoted the activity of calcified nodules by PDL cells.
These results suggest that laminin enhance the proliferation of undifferentiated PDL cells and remain them at undifferentiated stage and type I collagen support PDL cells to differentiate into osteoblasts or ceomentoblasts at later stage of regeneration. It seems that ECM controls the biological activities of PDL cells and plays an important role in periodontal tissue regeneration.

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We have previously reported that the retention fluid components of chronic sinusitis induced neutrophil adherence to microvascular endothelial cells, and this adherence was inhibited by anti-IL-lβ antibody.
In this study, we carried outin situhybridization experiments using the maxillary sinus mucosa to identify the cells responsible for the production of IL-1βin vivo.
IL-1β mRNA was noted in the part of PMNs which migrate extravascularly as well as in some macrophages in the maxillary sinus mucosa taken from the patients with chronic sinusitis. On the other hand, IL-1β mRNA was not noted in intravascular PMNs. The expressions of ICAM-1 and ELAM-1 were also investigated employing immunohistochemical techniques using anti-ICAM-1 and anti-ELAM-1 monoclonal antibody. ICAM-1 was strongly expressed upon the lumeninal surface of vascular endothelial cells. On the other hand, ELAM-1 was evident only in relatively shallow regions of the maxillary sinus mucosa.
These findings suggest that IL-1β produced by PMNs which migrate extravascularly and infiltrate into the maxillary sinus mucosa, and then stimulates the vascular endothelial cells to express the adhesion molecules such as ICAM-1 and ELAM-1. It is considered that this mechanism plays an important role in the repeated infiltration of PMNs which caused in a prolonged action of chronic sinus inflammation.

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The effect of direct irritant action of anti-inflammatory powder drugs (all 25mg) in canine gastric mucosa was studied using an endoscopic compressed air drug delivery system technique. Aspirin-induced mucosal lesions were limited to the target site of gastric mucosa in all dogs. Indomethacin (30%) and diclofenac Na (40%) induced superficial gastritis at the pyloric antrum, but did not appear to spray sites. Predonisolone caused the erosion and edema throughout the stomach. No lesions were induced by lactose and sucralfate.
In the present study, we could observe the experimental gastric mucosal lesion at the target site of the gastric mucosa by an administration of a small amount of powdered drugs via an endoscope. The administration of powdered drugs by endoscopy is useful for the investigation of direct effects of those on the gastric mucosa.

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The effect of several agents on mitogen-induced proliferative responsein vitrowas studied by using colorimetric MTT [3- (4, 5-dimenthyl-2-thiazolyl) -2, 5-diphenyl-2H tetrazolium bromide] assay. Immuno-modulator levamisole and TOK-8801 enhanced concanavalin A and lipopolysaccharide response in murine splenocytes at concentrations of 10^-6-10^-5M and 10^-4M, respectively. Immunosuppressant mizoribine decreased dose dependently this response.
These results show that MTT assay may be a useful method for the evaluation of immunological affecting agents on cell proliferative response.

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We studied the effect of Sho-seiryu-to and Saiboku-to on arachidonic acid metabolism by cultured porcine tracheal smooth muscle cells. HPLC analysis revealed that both cyclooxygenase derivatives, especially PGE₂, and lipoxygenase derivatives released in the culture medium were increased during 24-hour incubation of the tracheal smooth muscle cells with 1, 000μg/ml of Sho-seiryu-to. In contrast, the incubation of the tracheal smooth muscle cells with 1, 000μg/ml of Saiboku-to decreased both cyclooxygenase and lipoxygenase derivatives released in the culture medium. Increased release of PGE₂from tracheal smooth muscle cells incubated with Sho-seiryu-to may support the therapeutic benefit for bronchial asthma because of broncho-dilatation effect by PGE₂, and the suppression of the production of arachidonic acid metabolites by Saiboku-to may show the antiinflammatory action of Saiboku-to.

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