Rheumatoid arthritis (RA) is a refractory disease in most cases. Immunomodulators (IMs) such as gold sodium thiomalate (GST) or D-penicillamine (D-Pc) are sometimes very effetive for RA, and can induce a remission occasionally. The remission rate, however, is only 10-20% at most, and the duration of remission is a few years generally. Moreover, we have no more than 10 IMs now. Therefore, we can not induce a remission in most patients with RA, and can not maintain a remission for long time, resulting no significant difference in the long term outcome from the natural course.
In order to increase the remission rate and elongate the remission time with the limited number of IMs, we are trying the additive combination therapy of IMs. That is, to RA patients who are treated with an IM effectively but incompletely, another IM is added aiming at complete remission. We have had many cases of successfully treated with this method. For example, a patient controlled incompletely with 200 mg D-Pc, was induced to complete remission by addition of 100 mg tiopronine.
According to our multicenter clinical trials, it appears that the combination of GST and bucillamine is fitting, while the combination of GST or D-Pc with lobenzarit is rather unfitting.

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Intercellular adhesion molecule 1 (ICAM-1), one of the ligands for leukocyte function-associated antigen 1 (LFA-1), plays important roles both in inflammation and in immune responses.
In order to directly examine the role of ICAM-1/LFA-1 pathway in inflammatory process, we investi-gated the effect of in vivo administration of monoclonal antibodies on inflammatory models in rats. In adjuvant arthritis which is a chronic inflammatory model, both 1 A 29 (anti-rat ICAM-1) and WT-1 (anti-rat CD 11a) exerted a very strong suppressive effect on the development of arthritis. These monoclonal antibodies also suppressed the increase in serum sialic acid content in arthritic rats. Suppression of inflammatory reaction by monoclonal antibodies was also observed in rat carrageenan induced pleurisy. Flow cytometric analysis revealed that infiltrated neutrophils in pleural cavity expressed CD ha, CD 11b, CD 11c and CD 18. Administration of 1 A 29, WT-1 or WT-3 (anti-rat CD 18) significantly inhibited neutrophil extravasation into pleural cavity. These monoclonal antibodies also inhibited protein exudate into pleural cavity. However, OX 42, which was reported to recognize a common epitope shared by CD 11b and CD 11c, had no significant effect.
These results clearly indicate that ICAM-1/LFA-1 pathway is involved in the pathogenesis of adjuvant arthritis and have a significant role in neutrophil extravasation in carrageenan induced pleurisy.

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Cellular proteases in human T4⁺ lymphocytes play important roles in membrane fusion and internalization of HIV-1 and in virus replication. We observed a novel membrane-bound serine protease, named tryptase TL₂, in human T4⁺ lymphocytes, which specifically binds to the V 3 domain of HIV-1 gp 120. The V 3 domain of HIV-1, containing the central motif-GPGR-sequence that is homologous to the active site Kunitz-type inhibitors, is conserved in almost all HIV-1 isolates. Neutralizing antibody against the V 3 domain of HTLV-III_B, inhibitors of tryptase TL₂ with a-GPCR-sequence in their reactive site and synthetic peptides corresponding to the sequ-ences of the V 3 domains of various HIV-1 strains inhibited the binding of HIV-1 gp 120 to tryptase TL₂ and also inhibited syncytia formation induced by HIV-1. These findings suggest that tryptase TL₂ binds to gp 120 intracting with the V 3 domain.
Signal peptidase and HIV-1 envelope precursor processing protease in Golgi apparatus are required for HIV-1 replication. We purified the HIV-1 envelope precursor processing enzyme from Golgi apparatus of human T 4⁺ lymphocytes. The enzyme had a molecular mass of 25 kDa and converted gp 160 to gp 120 and gp 41 without any further processing, with high efficiency at a pH optimum of 6.5-7.0.

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While the pharmacology of nonsteroidal antiinflammatory drugs (NSAIDS) has been extensively studied, the precise mechanism as to how NSAIDS function at immunological level is not fully understood. Alminoprofen is a newer NSAIDS with characteristic antiinflammatory function by inhibiting cylooxygenase and phospholipase A₂ activity. Incubation of normal human mononuclear cells (MNC) with alminoprofen significantly inhibited IgG- and IgM-rheumatoid factor production in PWM-stimulated MNC. Alminoprofen also inhibited immunoglobulin production. On the other hand, alminoprofen did not inhibit IL-1 and TNF-a production by LPS-stimulated MNC.
The data suggest that the inhibition of rheumatoid factor production by activated MNC may contribute to the therapeutic role of alminoprofen in the management of immune dysfunction in rheumatoid arthritis patients although alminoprofen may differ from disease modifying antirheumatic drugs in immune regulation.

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Cholesteatoma otitis media is characterized by the accumulation of desquamating keratinized epithelium within the middle ear usually accompanied by intense bone destruction beneath the subepithelial granulation tissue. We have previously reported that surgically obtained cholesteatoma tissues (CT) produce the bone resorbing activity (BRA) attributable to interleukin (IL) -1α and prostaglandin (PG) E₂ in culture. The subsequent analysis revealed the presence of tumor necrosis factor (TNF) α in CT culture supernatants, however, the amounts of TNFα were not sufficient enough to induce bone resorption in mouse calvarial assay system. The monoclonal anti-bodies to IL-1α or TNFα were added into the CT culture to investigate the interaction between IL-1α and TNFα. It was found that the anti-TNFα antibody partially reduced the production of IL-1α and BRA from CT, whereas TNFα production was blocked remarkably by anti-IL-1α antibody. These results suggest that the generated IL-1α and TNFα may act as reciprocal accelerator (s) and that its self-perpetuation relates to the bone destruction in cholesteatoma otitis media.

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The adult respiratory distress syndrome (ARDS) occurs in patients with various underlying illnesses. It has been suggested that complement activation may play a crucial role in the pathophysiology of ARDS by direct actions of complement fragments and/or by release of mediators including endotoxin, superoxide anion, and arachidonate metabolites.
For clarification of mechanisms for ARDS, we studied functional changes of alveolar macrophages (AM) by using a septic lung model of rats after the operation of cecal ligation and puncture. Septic AM generated significantly less amounts of leukotriene B₄ (LTB₄), 12- and 5- hydroxyeicosatetraenoic acids (HETEs) by stimulation with A 23187 than the control AM did. Immunoreactive LTC₄ in the bronchoalveolar lavage fluids from the septic rats was more than that from the control rats. Therefore, it was speculated that the AM of endotoxemic rats released lipoxygenase metabolites in alveoli and then these AM obtained showed reduced 5-lipoxygenase activity. In contrast, these cells released enhanced amounts of O^-₂ on stimulation with PMA.
To examine effects of LPS on arachidonate metabolism and O^-₂ generation, relatively high dose of LPS was i.v administered in rats. The results indicated that AM were activated by the injected LPS to release increased amounts of O^-₂ and to show enhanced 5-lipoxygenase activity.
It is concluded that such the discrepancy of 5-lipoxygenase activity between AM from LPS-injected rats and those from endogenously endotoxemic rats remains an interesting riddle in the future study of ARDS.

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The effect of anti-ulcer agents, some antimicrobial agents and synthetic dyes on the growth of Helicobacter pylori was studied using 21 strains of our isolates and the standard strain of H. pylori NCTC 11638. The oldest orally active beta-lactam, penicillin V and recently developed macrolide, roxithromycin inhibited the growth at remarkably low concentrations, but the activities of two new quinolones were moderate. Also methylene blue showed moderate inhibitory activity but indigo carmine did not. Among the cytoproiective anti-ulcer agents, sucralfate showed no inhibitory activity, whereas the two terpenoid anti-ulcer agents, plaunotol and teprenone inhibited the growth, suggesting the possibility that the anti-H. pylori activity participates in the mode of action. The hypothesis is supported by the fact that plaunotol exerted bactericidal action even when contacted for 3 h at its minimal inhibitory concentration.

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We found inhibitors against myeloperoxidase (MPO) release from human polymorphonuclear leukocytes (PMN) . We designated them aseanostatins (AS), because these inhibitors were extracted from an actinomycete isolated in Thailand. AS were purified by a series of column chromatography of charcoal and silicagel, and HPLC. Physico-chemical chracterization by gas liquid chromatography and GC-MS indicated that two component P 1 and P 5 as pure forms were identical with i-14: 0 (12-methyltridecanoic acid) and ante-i-15: 0 (12-methyltetradecanoic acid), respectively. AS P 1 and P 5 showed IC₅₀ of 0.96 and 0.54 beg/ml to the MPO release, respectively. AS, however, did not inhibit β-glucuronidase release, but inhibited O^-₂ production a little. Moreover, those had no effects on other functions of PMN.
These findings suggest that AS may regulate MPO release or O^-₂ production which related to over reaction in inflammatory site.

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We have previously reported the stimulatory effect of leukotriene C₄ (LTC₄) and inhibitory one of eicosapentaenoic acid (EPA) on in vitro tube formation of endothelial cells. In this paper, we examined the precise mechanisms of the effects of both substances on tube formation.
When bovine endothelial cells were cultured between collagen gels, the cells were proliferated and formed a tube for two days. As our previous report showed EPA (5, μg/ml) -pretreatment caused a significant suppression of tube formation, the cell growth was examined in the collagen gel after EPA-pretreatment. The growth of endothelial cell pretreated with EPA was significantly inhibited. On the other hand, although the tube formation was stimulated by LTC₄, but not by LTD₄, the cell growth was not affected by LTC₄. However, the migration was stimulated by LTC₄. By the checker board analysis, LTC₄ stimulated chemokinesis slightly and chemotaxis significantly.
Finally, we examined the effect of LTC₄ on tube formation of EPA-pretreated endothelial cells. In EPA-enriched cells LTC₄ failed to stimulate the tube formation.
These data strongly suggested that EPA could be one of the inhibitors of inflammatory angiogenesis.

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Medullasin is a serine protease found in human bone marrow cells. It resembles elastase, but has no elastinolytic activity. It consists of 238 amino acid residues with Tie as the amino terminal and His as the carboxyl terminal. Medullasin causes inflammation characterized by the infiltration of a large number of macrophages when injected into animal skin. Protease inhibitors in plasma such as α₂-macroglobulin and α₁-antitrypsin inhibited the protease by equimolar ratio. Injection of the mixture of medullasin and α₂-macroglobulin or α₁-antitrypsin into animal skin caused no inflammation. When these inhibitors were injected 5 sec after the injection of medullasin, phlogistic activity of the protease was not prevented by them.
These results indicate that medullasin released into the tissues directly from granulocytes attaches to them immediately in such a manner that plasma inhibitors are not able to inhibit it. Therefore, medullasin released from granulocytes directly to the tissues is considered to be able to act on them in spite of the presence of a large amount of plasma protease inhibitors.

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Natural killer cells (NK) activity of normal, healthy donors against K 562 cell line were investigated with or without the supernatants of cultured peripheral blood lymphocytes from primary, nephrotic syndrome.
When the effector cells were incubated for three days with the supernatants of cultured PHA-stimulated lymphocytes from both untreated and treated patients, NK activity was significantly inhibited from normal donors.
Consequently, it was suggested that the inhibitory factor on NK activity consisted in the supernatants of cultured lymphocytes from pediatric patients with nephrotic syndrome.

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The pathogenic mechanism of fulminant hepatitis induced by a 400 mg/kg D-galactosamine was investigated in male Wistar rats. The extent of liver injury was assessed by measurements of serum transaminases, serum total bilirubin and relative liver weight at 24 hr after D-galactosamine administration.
WF 11605 is a selective antagonist of leukotriene B₄ (LTB₄) . The compound was isolated as a product of fungal metabolite. WF 11605 inhibited LTB₄-induced chemotaxis of rabbit polymorphonuclear leukocyte (PMN) with an IC₅₀ value of 1.7 × 10^-7 M and blocked ³H-LTB₄ binding to PMN membrane at 5.6 × 10^-6 M (IC₅₀) . WF 11605 also inhibited LTB₄-induced degranulation of rabbit PMN at 3.0 × 10^-6 M (IC₅₀) . However, the compound did not show any inhibitory effect on platelet activating factor (PAF) -and N-formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP) -induced degranulation at concentrations up to 10^-4 M. In order to determine whether LTB₄ contributes to the occurrence of the hepatitis in rats, we therefore attempted to prevent the hepatitis by WF 11605. Intraperitoneally administered WF 11605 at doses of 10, 32 and 100 mg/kg significantly prevented the increase of serum transaminases and bilirubin in a dose dependent manner. We conclude from our results that LTB₄ might be involved in the pathogenesis of D-galactosamine-induced hepatitis in rats.

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The therapeutic effects of IPD-1151 T, a new anti-allergic agent, were compared with those of Tranilast in 356 patients with allergic rhinitis. The daily dosage was 300 mg (one capsule each, t.i.d.) for IPD-1151 T and 300 mg (one capsule each, t.i.d.) for Tranilast. Both test drugs were orally given for 6 weeks.
The results of this study are summarized as follows :
(1) IPD-1151 T showed significant superiority to Tranilast in “final global improvement rating”. Improvement rating was 55.0% for IPD-1151 T and 37.8% for Tranilast in terms of “moderate” plus “marked” improvement category.
(2) The item evaluated, “overall safety rating” showed significant superiority of IPD-1151 T to Tranilast. The incidence of an adverse reaction or abnormal laboratory finding was noted as 3.7% fo IPD-1151 T and 11.0% of Tranilast.
(3) The item evaluated, “global utility rating” showed significant superiority of IPD-1151 T to Tranilast. The utility rating was calculated as 53.2% of IPD-1151 T and 34.7% of Tranilast in terms of “moderately” plus “markedly” useful category.
(4) Side effects were observed in 6 patients (3.7%) of IPD-1151 T and 18 patients (11.0%) Tranilast. All of these symptoms disappeared by symptomatic treatment or discontinued test drug.
All the above results indicate that IPD-1151 T is safer and more effective than Tranilast. Therefore, IPD-1151 T is judged as a highly useful anti-allergic agent for the treatment of allergic rhinitis.

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