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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We compared TS-110 4 mg, 3times daily with a control drug, diclofenac sodium 25 mg, 3times daily in a double-blind study in order to evaluate clinical utility of TS-110 against lumbago-related disease, cervico-brachial syndrome and scapulohumeral periarthritis.
1. Concerning the final overall improvement rating, the improvement rate (the percentage of the patients graded as“inproved”or better) was 60.8% (76/125) and 55.7/ (73/131) in Groups T and D, respectively, suggesting the same level of efficacy in Groups T and D.
2. As to the global safety rating, the safety rate (the percentage of patients in whom the drug was graded as“safe”) was 77.8% (112/144) and 73, 4% (116/158) in Groups T and D, respectively.
3. With respect to utility rating, the utility rate (the percentage of patients in whom the drug was graded as“useful”or better) was 58.1% (72/124) and 46.6% (61/131) in Groups T and D, suggesting the same level of usefulness in Groups T and D.
The utility rate obtained by ITT analysis was 56.2% (82/140) and 42.7% (70/164) in Groups T and D, respectively, with the utility rate in Group T significantly higher than that in Group D (p=0.0178) .
4. A total of 38 cases of adverse reactions were noted in 32 of 148 patients (21.6%) in Group T, and 68 cases in 46 of 164 patients (28.0%) in Group D. As to the breakdown of symptoms, 27 and 38 cases of gastrointestinal symptoms were noted in Groups T and D, respectively, one case of drug eruption was noted in Group D, two and ten cases of symptoms in the mental and nervous systems in Groups T and D, respectively, and five and seven cases of other adverse reactions in Groups T and D, respectively. Four and 12 cases of abnormal clinical laboratory test values in Groups T and D, respectively, were judged to be adverse reactions. None of these adverse reactions, however, were severe.
In light of these findings, TS-110 was judged to have the same level of efficacy and safety as diclofenac sodium against lumbago-related diseases, cervicobrachial syndrome and scapulohumeral periarthritis, and thus, to be a clinically highly useful drug.

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We compared TS-110 with a control drug, loxoprofen sodium, in a double-blind study for the purpose of objective evaluation of the clinical utility of TS-110 against postoperative pain and post-traumatic pain.
1. Primary evaluation parameters
1) Final overall improvement rating
The improvement rate (the percentage of the patients graded as “improved” or better) was 87.9% (51/58) and 85.7% (54/63) in Groups T and L, respectively, among patients with postoperative pain, and was 76.4% (42/55) and 76.1% (35/46) in Groups T and L, respectively, among patients with post-traumatic pain, revealing no significant differences between the groups.
2) Global safety rating
The study drugs were graded as “safe” in 88.7% (63/71) and 89.6% (69/77) in Groups T and L, respectively, among patients with postoperative pain, and in 86.9 (53/61) and 92.6% (50/54) in Groups T and L, respectively, among patients with post-traumatic pain, revealing no significant differences between the groups.
3) Utility rating
The utility rate (the percentage of patients in whom the drug was graded as“useful”or better) was 86.2% (50/58) and 82.5% (52/63) in Groups T and L, respectively, among patients with postoperative pain, and 67.91 (38/56) and 72.3% (34/47) in Groups T and L, respectively, among patients with post-traumatic pain, revealing no significant differences between the groups.
2. Secondary evaluation parameters
1) The time of apperrance of the effects after the first administration was 12.1% (7/58) and 17.5% (11/63) within 15 minutes, 34.5% (24/58) and 60.3% (38/63) within 30 minutes, 56.9% (33/58) and 76.2% (48/63) within 45 minutes, 89.7% (52/58) and 87.3% (55/63) within one hour, in Groups T and L, respectively, among patients with postoperative pain, and was 7.3% (4/55) and 6.5% (3/46) within 15 minutes, 23.6% (13/55) and 39.1% (18/46) within 30 minutes, 34.5% (19/55) and 45.7% (21/46) within 45 minutes, 69.1% (38/55) and 65.2% (30/46) within one hour, in Groups T and L, respectively, among patients with post-traumatic pain.
2) Concerning self-evaluations of the effects of study drugs made by patients, the percentage of patients who graded“unproved”or better on the 1st day was 80.7% (46/57) in Group T and 59.7% (37/62) in Group L, respectively, with the percentage in Group T significantly higher than in Group L (p<0.05) .
3) Adverse reactions were noted in 11.3% (8/71) and 10.4% (8/77) of patients with postoperative pain in Groups T and L, respectively. There were abdominal discomfort, GPT elevation and so tin, in Group fi, and sleepiness, GPT elevation and so on, in Group L Among patients with post-traumatic patients the incidences of adverse reactions were 131% (8/61) and 74% (4/54) in Groups T and L, respectively. There were abdominal pain and so on, in Group T, and GPT elevation in Group L.
In light of these findings, TS-110 was judged to be a clinically useful drug with immediate and sustained effects against postoperative pain and post-traumatic pain.

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