Glomerular infiltration by neutrophils is a hallmark of acute glomerulonephritis. The pathophysiological role of interleukin-8 (IL-8), a potent neutrophil chemotactic cytokine (chemokine), was explored in an animal model of acute immune complex-mediated glomerulonephritis by administering a neutralizing antibody against IL-8. Repeated injection of bovine serum albumin (BSA) into rabbits caused the deposition of immune complexes consisting of BSA and rabbit IgG in glomeruli. Histological analyses revealed a small but significant number of neutrophils in glomeruli and the fusion of epithelial cell foot processes. Concomitantly, urinary levels of protein and albumin increased markedly (3.20±0.97 and 1.39±0.53 mg/hr, respectively) compared with those of untreated animals (0.77±0.21 and 0.01±0.01 mg/hr, respectively) . Anti-IL-8 antibody treatment decreased the number of neutrophils in glomeruli by 40% and dramatically prevented the fusion of epithelial cell foot process. Furthermore, treatment with anti-IL-8 antibody completely normalized the urinary levels of protein and albumin (0.89±0.15 and 0.02±0.01 mg/hr, respectively) . These results indicated that IL-8 participated in the impairment of renal functions in experimental acute immune complex-mediated glomerulonephritis through activating as well as recruiting neutrophils. Here, we will overview the roles of chemokine in human diseases and discuss our therapeutic approaches to intervene inflammation targeting chemokine.

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Periodontitis is infectious disease by periodontopathic bacteria and characterized by gingival connective tissue loss, periodontal pocket formation and alveolar bone resorption. Unbalance of local immune responses may cause onset and progression of periodontitis. Gingival crevicular fluid (GCF) from the pocket of periodontitis patients contains immuno-competent cells, namely, polymorphonuclear leukocytes, lymphocytes, and macrophages.
Cell surface expression of Fcγ receptor II, III in GCF-polymorphonuclear leukocytes decreased in contrast to increase of complement receptor 3 as compared with those of autologous peripheral blood-polymorphonuclear leukocytes. Transcript levels of Fcγ receptor III and complement receptor 1, 3 significantly decreased. These phenomena may induce impaired phagocytosis of GCF-polymorphonuclear leukocytes and progression of periodontitis.
Regarding for GCF-lymphocytes, decreased T cell/B cell ratio and activated helper T cells and B cells were observed as compared with autologous peripheral blood-lymphocytes. ICAM-1 positive pocket epithelium and LFA-1 expression of GCF-lymphocytes may be related to cell exudation from connective tissue to gingival crevice.
HIV gag protein p24 positive macrophages in GCF increased dramatically in AIDS patients with periodontitis. PCR and RT-PCR of GCF-leukocytes yielded positive signals of p 24, demonstrating viral integration and production. These findings could be considered as a within-mouth source of viral reservoir.
These characteristics of immuno-competent cells in GCF may reflect local immune responses of inflammatory gingival tissues.

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It has been postulated that disturbances of ionic homeostasis may determine the vulnerability of the heart to reperfusion-induced injury, and that reactive oxygen species produced during reperfusion, cause oxidant stress to membrane protein or lipid that leads to disturbances of ionic homeostasis and arrhythmias. To test this hypothesis, we studied the efficacies of various reactive oxygen species scavengers in reducing ischemia-reperfusion (I/R) -induced myocardial injury in isolated perfused rat hearts. The effect of in vitro free radical generating system consisting of hypoxanthine (hyX) and xanthine oxidase (XO) on sarcoplasmic reticulum (SR) function was also investigated. In I/R hearts, the contractile function and coronary flow were reduced; reperfusion with histidine or a cocktail of superoxide dismutase (SOD) -catalase-mannitol resulted in significant protection against the effect of I/R. The incidence of arrhythmias during reperfusion was 100% in I/R hearts; the duration of arrhythmias was shortened with histidine or with SOD-catalase-mannitol. The decreased duration of normal sinus rhythm in I/R hearts was also protected with histidine or SOD-catalase-mannitol. The SR function assessed by oxalate-supported Ca²⁺ uptake rate in cell free preparations in the presence or absence of ruthenium red, a selective SR Ca²⁺-release channel blocker, was depressed by exposure to hyX-XO reaction. The observed effect of hyX-XO was SOD-inhibitable and was protected by SOD-catalase-mannitol. In samples where the Ca²⁺-release channel was blocked with ruthenium red, no changes in Ca²⁺ uptake rates were noted after ischemia only; the Ca²⁺ uptake rates in the presence of ruthenium red decreased in samples from ischemia-reperfused hearts, suggesting Ca²⁺-release channel dysfunction caused by oxygen-derived free radicals generated during reperfusion. Exposure of isolated heavy SR to hyX-XO reaction produced potent increase in Ca²⁺ efflux; the effect of hyX-XO reaction was protected by SOD or by established optimal conditions for specific closure of heavy SR Ca²⁺-release channel by ryanodine. Based on these lines of results, we conclude that the action of reactive oxygen species (possibly, superoxide radicals or singlet molecular oxygen) may be mediated in part through SR Ca²⁺-release channel, thereby causing Ca²⁺ overload linked to arrhythmogenic electrophysiological changes.

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Defensin has been known as a major antimicrobial component of neutrophil granules. Recently, we have purified a novel cationic antibacterial polypeptide of 11 kDa (CAP11) from guinea pig neutrophilis. In this study, we have compared the biological activity and gene expression between CAP11 and defensin. When stimulated, CAP11 was extracellularly released from neutrophils, accompanied by the release of a specific granule component (lysozyme), whereas defensin was released, accompanied by the release of an azurophil granule component (β-glucoronidase) . Defensin modulated neutrophil functions such as adhesion, phagocytosis and superoxide anion generation. However, CAP11 did not affected the neutrophil functions. Both CAP11 and defensin possessed histamine-releasing activity for mast cells, but CAP11 was 10-fold less potent than defensins. CAP11 exhibited 8-fold more antibacterial activity against Escherichia coli than defensin, and the activity was retained even in the presence of physiological concentration of NaCl, although the activity of defensin was completely lost in the presence of NaCl. Sequence analysis of CAP11 cDNA showed that pre-prosequence of CAP11 was similar to the sequences of cathelicidin family polypeptides. Furthemore, in situ hybridization study revealed that CAP11 mRNA was highly expressed in metamyelocytes among bone marrow cells. In contrast, defensin mRNA was expressed in promyelocytes and myelocytes. Together these observations indicate that CAP11, a member of cathelicidin family, which is synthesized during the late stage of neutrophil maturation and stored possibly in the specific granules, is released from stimulated neutrophils and functions as an antibacterial molecule in the extracellular milieu, whereas defensin released from the azurophil granules likely participate in the modulation of neutrophil functions and mast cell histamine release.

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The effect of a monoclonal antibody against IL-4 (11B11) on oral tolerance was investigated. Oral tolerance was induced by feeding mice with hen egg lysozyme (HEL) before immunization with HEL. 11B11 was ip injected 30 min before oral administration of HEL. The results showed that the oral administration of HEL suppressed immune responses to the antigen including DTH responses, production of both isotypes of IgG 1 and IgG2a antibodies, and proliferation of lymph node cells in a dose-dependent manner. These suppression were associated with a marked reduction of IFN-γ secretion and a partial decrease in IL-4 production by lymphoid cells. The treatment of the tolerant animals with 11B11 significantly blocked the suppression of DTH responses, IgG2a antibody production, and proliferative responses as well as IFN-γ secretion. In contrast, the monoclonal antibody facilitated the suppression of anti-HEL IgG1 antibody production and IL-4 secretion. Thus, oral antigens appear to induce not only Th1 but also Th2 cell tolerance and the neutralizing IL-4 with anti-IL-4 antibodies seems to reverse the suppression of Th1 cell responses, while the antibodies further reduce Th2 cell responses in oral tolerance, indicating the implication of IL-4 in the oral tolerization of Th1 cells.

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Tumor necrosis factor (TNF) -α is present in synovial fluid of patients with rheumatoid arthritis. It induces the expression of chemotactic cytokines (chemokines) such as interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) in synovial cells.
Based on our recent finding that reactive oxygen intermediates play important roles in mediating TNF-α action, we examined the effect of an antioxidant flavonoid, quercetin, on TNF-α-induced expression of IL-8 and MCP-1 in cultured human synovial cells. Quercetin suppressed the TNF-α-induced increase in the mRNAs for IL-8 and MCP-1 in a dose dependent manner. H₂O₂-mediated induction of these genes was also inhibited by quercetin. Electrophoretic mobility shift assays revealed that quercetin inhibited the activation of NF-κB by TNF-α.
These results demonstrate for the first time that quercetin suppresses TNF-α-mediated stimulation of IL-8 and MCP-1 expression, at least in part, by inhibiting the activation of NF-κB.

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慢性関節リウマチに対するTS-110長期投与時の臨床的有用性をオープン試験により検討した.
1.総投与例数は134例であり, 12週以上投与108例 (80.6%) , 24週以上投与82例 (61.2%) , 48週投与30例 (22.4%) であった.
2.各臨床評価項目を各症例の最終評価日において試験開始時と比較し, 疼痛および腫脹関節数, 関節点数, 疼痛点数, 日常生活動作の上肢, Lansburyのactivity indexで有意な改善が認められた.
3.患者の満足度における「満足」以上の割合は, 24週後では16.2% (12/74例) , 32週後以降はいずれも20%以上でほぼ一定であり, 48週後では26.7% (8/30例) であった.
また, 医師の症状改善度評価における「改善」以上の割合は, 24週後では33.3% (25/75例) , 24週後以降はいずれも30%以上であり, 48週後では37.9% (11/29例) であった.
4.全般改善度における「改善」以上の改善率は, 24週では33.8% (25/74例) , 48週では40.0% (12/30例) であり, 最終評価日では30.5% (36/118例) であった.
5.副作用は, 133例中41例 (30.8%) で64件認められ, いずれも軽度または中等度であった.なお, 治験薬剤との関連性が否定できない臨床検査値の異常変動は, いずれも副作用として扱われた.
発現率は投与期間に伴い高くなることはなく, また, 副作用の種類においても, 投与期間が長くなることに伴い, 特に異なるものは認められなかった
6.概括安全度における「安全である」の症例の割合 (安全率) は, 24週では85.3% (64/75例) , 48週では90.0% (27/30例) , 最終評価日では69.2% (92/133例) であった.
7.有用度における「有用」以上の有用率は, 24週では36.5% (27/74例) , 48週では43.3% (13/30例) , 最終評価日では26.8% (34/127例) であった.
これらのことから, TS-110の慢性関節リウマチに対する長期投与における有効性面, 安全性面はいずれも良好であり, 臨床的に有用な薬剤であると考えられる.

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慢性関節リウマチに対するTS-110の臨床的有用性を評価するため, インドメタシンを対照薬として, TS-110 1日12mg分3 (T群) , インドメタシン75mg分3 (I群) で6週間経口投与し, 二重盲検法により比較検討した.
(1) 総投与例は189例 (T群95例, I群94例) であり, ITT対象症例は有効性, 安全性, 有用性いずれも183例 (T群91例, I群92例) であった.
(2) 背景因子には「性別」 (p=0.0566) , 「併用薬の有無」 (p=0.0313) 以外, 偏りは認められなかった.
なお, 本偏りの改善度に及ぼす影響についてMantel-Haenszel法などにて検定を行った結果, 影響は認められなかった.
(3) 最終全般改善度において, 「改善」以上の改善率は, T群, I群それぞれ29.7% (27/91例) , 17.4% (16/92例) であり, T群の有効性は, I群と同等であった.
(4) 副作用発現率は, T群, I群それぞれ19.8% (18/91例) , 26.1% (24/92例) であり, 症状別では消化器症状がT群で13件, I群で17件, 皮疹, 〓痒感の過敏症がT群で1件, I群で1件, めまい, 頭痛などの精神・神経系の症状がT群で1件, I群で11件, 臨床検査値の異常変動がT群で11件, I群で7件に認められた.
(5) 概括安全度において, 「安全である」の安全率はT群, I群それぞれ80.2% (73/91例) , 73.9% (68/92例) であり, T群の安全性は, I群と同等であった.
(6) 有用度において, 「有用」以上の有用率はT群, I群それぞれ28.6% (26/91例) , 15.2% (14/92例) であり, T群がI群より有意 (p=0.0289) に高かった.
(7) その他副次評価項目では, 最終評価日における「満足」以上の患者の満足度はT群がI群より有意 (p=0.0166) に高かった.
以上より, TS-110の12mg/日投与は慢性関節リウマチに対し, インドメタシン75mg/日投与と同等の有効性, 安全性を有し, 患者の満足度を反映した評価では有用性の高い薬剤であると判断された.

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