The tyrosine kinase inhibitors erbstatin and herbimycin A inhibited the chemotactic response and the superoxide anion (O^-₂) production stimulated by FMLP in human polymorphonuclear leukocytes (PMN) in similar manners. These compounds also inhibited phospholipase D (PLD) -catalysed breakdown of phosphatidyl choline, suggesting a possible link between tyrosine kinase and PLD. In the presence of propranolol (phosphatidic acid (PA) phosphohydrolase inhibitor), or ethanol, the activation of PLD results in the modulation of PA and/or diglyceride (DG) generation, producing an irregularity in O^-₂ production. However, PMN motility was not affected in these conditions.
These results suggest that PLD is a downstream effector of FMLP-induced tyrosine kinase activation that leads to activation of the PMN superoxide release but not to chemotactic migration. In contrast, the tyrosine kinase inhibitors did not inhibit inositol 1, 4, 5-triphosphate generation and increase of intracellular concentration of free calcium. These results support the premise that there are divergent signaling pathways for activations of distinct enzymes and functions in PMN. This premise has been proved by the recent renewed appreciation for the complexities of chemoattractant signaling pathways.

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Vaccines against cholera and other enteric diseases would be a major benefit for developing countries where these diseases are often life threatening, especially among children. Enterotoxigenic Escherichia coli (ETEC) causes acute watery diarrhea by colonizing the small intestine and producing heat-stable and heat-labile enterotoxins (ST and LT) . The LT-B, a binding subunit of E. coli LT is a highly active oral immunogen. The aim of this study was to understand molecular and cellular mechanisms for the induction and regulation of LT-B-specific immune responses following oral immunization.
We initially assessed E, coli LT as mucosal immunogen and as adjuvant in mice. Oral administration of LT to BALB/c (H-2^d) mice induced high mucosal IgA and serum IgG antibody responses. Analysis of LT-B-specific CD4⁺ T helper (Th) cells from Peyer's patches (PP) or spleen (SP) revealed a mixed Thl (IFN-γ) and Th2 (IL-4 and IL-5) cell pattern.
The B cell and Tcell epitopes of LT-B which induce effective secretory IgA (S-IgA) responses were examined in the next experiment. LT-B-specific S-IgA of BALB/c mice orally immunized with recombinant LT-B expressed in S. typhimurium showed major recognition of peptide starting at residue 36. PP CD4⁺ T cells cultured with the peptide covering residues 34 to 43 of the LT-B molecule produced large amounts of IL-5 and IFN-γ. These findings indicated that the residues 34 to 43 of the LT-B molecule enhanced the production of both Thl-and Th2-type cytokines. According to these findings, a peptide [peptide (26-45) ] encompassing both the B-cell and T-cell epitopes was synthesized as a possible candidate for oral vaccine. When BALB/c (I-A^d) mice as well as three strains of B10 congenic mice [B10 (I-A^b), B10. D2 (I-A^d), and B10. BR (I-A^k) ] were orally immunized with the peptide, serum IgG and S-IgA responses were elicited not only to the peptide itself, but also to the native LT. In addition, PP CD4⁺ T cells from all the stains of B10 congenic mice as well as BALB/c mice induced strong peptide-specific T cell proliferation and cytokine synthesis, suggesting the promiscuous binding of the peptide to MHC class II molecule.
A possibility for induction of oral tolerance by mucosal administration with the LT-B peptide was examined by a multiple low dose feeding of the immunogenic peptide (26-45) . Following the multiple oral feeding with the LT-B peptide induced systemic unresponsiveness in BALB/c mice resulting in diminished serum IgG responses and maintenance of appropriate mucosal IgA responses. CD4⁺ T cells from SP of the tolerized mice failed to proliferate, whereas PP CD4⁺ T cells responded to the peptide. RT-PCR revealed that the PP CD4⁺ T cells expressed significant levels of mRNA for IFN-γ, IL-2, IL-4, and TGF-β. These results indicated that SP CD4⁺ T cells induced a state of systemic T cell anergy.

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Concentration of Indometacin farnesil (IMF), indomethacin (IND) transformed from IMF in serum and synovial fluid were studied after single oral administration of Infree^® capsule in patients with rheumatoid arthritis (RA) . Samples of blood, synovial fluid were taken 3 hours, 6 hours, 9 hours and 12 hours after Infree^® administration.
1. The maximam concentration of IMF, IND transformed IMF in serum and synovial fluid were obtained at 9 hours after Infree^® administration.
2. The ratio of IND/IMF in synovial fluid is constant and higher level than that of IND/IMF in serum. The ratio of IND/IMF in serum is lowered each sampling hours.
These results suggested that Infree^® capsule has a constant analgesic/antiinflammatory effect by a good intra-articular transformation of IMF in inflammatory joints of RA patients.

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Objective: Administration of mercuric chloride (HgCl₂) to Brown Norway (BN) rats causes autoimmune glomerulonephritis and necrotizing vasculitis in the gut. In this study we analyzed the effects of FK506 on tissue injury and autoantibody production in HgCl₂-treated BN rats.
Methods: FK506 was administered during the“early phase” (days 1 to 10, concurrently with HgCl₂) or the“late phase” (days 11 to 14) . The autoantibody titers and urinary protein levels were mesured serially during the experimental period. Histopathological grade of tissue injury was assessed after the rats were sacrificed on day 15.
Results: HgCl₂-treated BN rats showed interstitial pneumonitis in the lung, necrotizing vasculitis in the gut and glomerulonephritis. The titers of anti MPO and anti DNA antibodies increased to a maximum on day 10. FK506 given during the early phase significantly inhibited autoantibody increases, pneumonitis and proteinuria. On the other hand, FK506 given during the late phase exacerbated the vasculitis in the gut but slightly lowered the titer of anti MPO antibodies.
Conclusion: These results indicate that the effect of FK506 depends on the time of administration, or the pathological state at the time of treatment. Early administration of FK506 as a combination treatment may prevent necrotizing vasculitis caused by immunological mechanisms.

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Diffuse panbronchiolitis (DPB) is characterized by a dense neutrophil infiltration in association with the high level of interleukin 8 (IL-8) in the airway and an accumulation of lymphocytes, especially CD8⁺HLA-DR⁺cells. In this study, we demonstrated that IL-8 mRNA was expressed in alveolar macrophages in the air space, bronchiolar epithelial cells and endothelial cells by in situ hybridization method, while not in foamy macrophages in the interstitium. A two-color analysis of T cell subsets in bronchoalveolar lavage (BAL) fluid also revealed a significant increase in the number of CD8⁺CD11b^- cytotoxic T cells and CD4⁺CD29⁺ memory T cells and the mean fluorescence intensity of CD49d (very late antigen, VLA-4α) on CD4⁺cells in the patients. Furthermore, the level of RANTES and MIP-1α in DPB patients was significantly higher than that in healthy volunteers, and there was a trend toward a correlation between RANTES concentration and the absolute number of memory T cells in BAL fluid (r=0.731, p=0.188) .
Our results suggest that elevation of memory T cells and cytotoxic T cells, in addition to neutrophils, in the airway lumen of DPB patients may contribute to chronic bronchial inflammation, possibly through up-regulation of adhesion molecule, VLA-4 and chemokines produced in the lung.

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The tumor antigenic peptides of human melanomas have been determined, and applied in the clinical trials. It is also important to investigate the biochemical process for producing these antigenic peptides in the cells. In this report, we suggested that 70 kDa heat shock protein (hsp 70) makes physical complex with TAP molecules. Futhermore, hsp 70 could play an important role to feed HLA-A2-restricetd melanoma tyrosinase antigenic peptides into ER via TAP molecules.

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腰痛性疾患, 頸肩腕症候群, 肩関節周囲炎に対するTS-110の臨床的有用性を評価するため, ジクロフェナクナトリウムを対照薬として二重盲検法により比較検討した.投与量は1日あたりT群が12mg, D群が75mg, いずれも1日3回毎食後投与とし, 投与期間は2週間とした.解析については「鎮痛消炎剤の臨床評価方法に関するガイドライン (1985.5) 」に則り, 腰痛性疾患, 頸肩腕症候群, 肩関節周囲炎の3疾患全体で行った.
1.総投与例数331例 (T群158例, D群173例) のうち, 解析対象例は, 有効性256例 (T群125例, D群131例) , 安全性302例 (T群144例, D群158例) , 有用性255例 (T群124例, D群131例) であった.
2.最終全般改善度では改善率 (改善, 以上) は, T群, D群それぞれ60.8% (76/125) , 55.7% (73/131) であり, T群の有効性はD群と同等であった.疾患別では, 腰痛性疾患ではそれぞれ68.2% (30/44) , 60.8% (31/51) , 頸肩腕症候群では57.9% (22/38) , 47.6% (20/42) , 肩関節周囲炎では55.8% (24/43) , 57.9% (22/38) であった.
3.概括安全度では安全率 (安全である) が, T群, D群それぞれ77.8% (112/144) , 73.4% (116/158) であり, T群の安全性はD群と同等であった.疾患別では腰痛性疾患でそれぞれ84.3% (43/51) , 73.3% (44/60) , 頸肩腕症候群で70.8% (34/48) , 75.0% (39/52) , 肩関節周囲炎で77.8% (35/45) , 71.7% (33/46) であった.
4.有用度では有用率 (有用, 以上) は, T群, D群それぞれ58.1% (72/124) , 46.6% (61/131) であり, T群の有用性はD群と同等であった.疾患別では腰痛性疾患でそれぞれ68.2% (30/44) , 50.0% (25/50) , 頸肩腕症候群で52.6% (20/38) , 41.9% (18/43) , 肩関節周囲炎で52.4% (22/42) , 47.4% (18/38) であった.
5.副作用はT群では148例中32例 (21.6%) 38件に, D群では164例中46例 (28.0%) 68件に認められ, 症状別では消化器症状がT群で27件, D群で38件, 薬疹がD群で1件, 精神・神経系の症状がT群で2件, D群で10件, その他の副作用がT群で5件, D群で7件, 副作用とした臨床検査値異常がT群で4件, D群で12件に認められた.
以上の結果より, TS-110は腰痛性疾患, 頸肩腕症候群, 肩関節周囲炎に対しジクロフェナクナトリウムと同等の有効性, 安全性を有し, 臨床的に有用性の高い薬剤であると考えられた.

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手術後疼痛および外傷後疼痛に対するTS-110の臨床的有用性を客観的に評価するため, ロキソプロフェンナトリウムを対照薬として二重盲検法により比較検討した.
1.患者背景
手術後疼痛においては, 有効性解析対象症例は121例 (T群58例, L群63例) , 安全性解析対象症例は148例 (T群71例, L群77例) , 有用性解析対象症例は121例 (T群58例, L群63例) であり, 外傷後疼痛においては, 有効性解析対象症例は101例 (T群55例, L群46例) , 安全性解析対象症例は115例 (T群61例, L群54例) , 有用性解析対象症例は103例 (T群56例, L群47例) であった.
患者背景において, 両群間に手術後疼痛では, 「性別」, 「体重」, 「既往症の有無」, 「合併症の有無」, 「麻酔法」, 「感染の有無」, 観察項目の初期値の「局所熱感」, 外傷後疼痛では「受傷から来院までの時間」以外偏り (p<0.15) のあった項目は認められなかった.なお, これらの偏りの主要評価項目に及ぼす影響をMantel-Haenszel法により検討したが, 影響は認められなかった.
2.主要評価項目
1) 最終全般改善度
改善率 (改善, 以上) は, 手術後疼痛ではT群87.9% (51/58) , L群85.7% (54/63) , 外傷後疼痛ではT群76.4% (42/55) , L群76.1% (35/46) であり, いずれも両群間に有意差は認められなかった.
2) 概括安全度
「安全である」は, 手術後疼痛ではT群88.7% (63/71) , L群89.6% (69/77) , 外傷後疼痛ではT群86.9% (53/61) , L群92.6% (50/54) であり, いずれも両群間に有意差は認められなかった.
3) 有用度
有用率 (有用, 以上) は, 手術後疼痛ではT群86.2% (50/58) , L群82.5% (52/63) , 外傷後疼痛ではT群67.9% (38/56) , L群72.3% (34/47) であり, いずれも両群間に有意差は認められなかった.
3.副次評価項目
1) 初回投与後の効果発現時間
初回投与後の効果発現時間は, T群, D群それぞれ, 手術後疼痛においては15分以内12.1% (7/58) , 17.5% (11/63) , 30分以内34.5% (20/58) , 60.3% (38/63) , 45分以内56.9% (33/58) , 76.2% (48/63) , 1時間以内89.7% (52/58) , 87.3% (55/63) であり外傷後疼痛においては, それぞれ15分以内7.3% (4/55) , 6.5% (3/46) , 30分以内23.6% (13/55) , 39.1% (18/46) , 45分以内34.5% (19/55) , 45.7% (21/46) , 1時間以内69.1% (38/55) , 65.2% (30/46) であった.
2) 初回投与後の効果持続時間
初回薬剤服薬後の鎮痛効果持続時間は, 手術後疼痛・外傷後疼痛いずれにおいても両群ともほぼ同様の分布であった.
3) 臨床症状の推移
開始時と最終評価日の臨床症状の推移は, いずれの項目も両群ともほぼ同様の推移であった.
4) 症状改善度 (患者の自己評価)
試験薬剤の効果に関する1日後, 2日後および3日後の患者の自己評価は, 手術後疼痛では, 1日後の「良くなった」以上でT群80.7% (46/57) , L群59.7% (37/62) であり, T群がL群に比し有意 (p<0.05) に高かった.外傷後疼痛では, 両群ともいずれの時期においても同様の値であった.
5) 症状改善度 (医師の評価)
1日後, 2日後および3日後に評価した各臨床症状の改善度は, 手術後疼痛, 外傷後疼痛いずれにおいても両群のいずれの時期においても同様の値であった.
6) 副作用
手術後疼痛の副作用はT群に11.3% (8/71) , L群に10.4% (8/77) 認められ, 内訳はT群では腹部不快感3件, 腹痛, 悪心, 眠気各1件およびGOTの上昇1件, GPTの上昇2件, またL群では眠気2件, 腹部不快感, 下痢, 掻痒感, 耳閉感各1件およびGOTの上昇1件, GPTの上昇2件であった.外傷後疼痛の副作用はT群に13.1% (8/61) , L群に7.4% (4/54) 認められ, 内訳はT群では腹痛3件, 腹部不快感, 悪心, 下痢, 発疹, ふらふら感, 肝機能異常, 動悸各1件, またL群では眠気, 肝機能障害各1件およびGOTの上昇1件, GPTの上昇2件であった.
4.ITT解析
手術後疼痛・外傷後疼痛いずれにおいてもPC解析とほぼ同様の結果であった.
以上のことから, TS-110は手術後疼痛, 外傷後疼痛に対し効果の発現時間は短く, 持続性もあり臨床的に有用な薬剤と判断した.

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