No. 6: color of serum, No. 7: spontaneous hemolysis normal range: U/ml
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1 Key words: complement, immune complex, solubilization, acute serum sickness, chronic serum sickness
2 No. 6: color of serum, No. 7: spontaneous hemolysis normal range: U/ml
3 生体 内 にお け る補体 に よる免 疫複合 体 の可溶 化 生 検 は エ ー テ ル麻 酔 後,片 腎 を摘 出 し,そ の 後, 頸 静 脈 よ り採 血 を施 行 した.片 表2 Score of 1053 immune complex in ASS- nephritis 腎摘 出 及 び採 血 の た め 大 部 分 の ラ ッ トは 死 亡 し た た め 同 一 ラ ッ トで の 腎 生 検 は 施 行 で きな か っ た.腎 FITC標 識-抗BSA,-抗 体,-抗 ラ ッ トC3ヤ 染 色 後,蛍 切 片 は, ラ ッ トIgGウ サ ギ抗 ギ抗 体(各Cappel社)で 光 顕 微 鏡 に て 糸 球 体 へ のICの 沈着 を観 察 した.(図2) 2) ラ ッ トの 補 体 価(CH50), C3及 びCRA の測 定 ラ ッ ト補 体 価(CH50)の て 測 定 し, CVF, 測 定: Mayer法 に TO: turpentine oil,cvf: *p<0.05, **p<0.02 Cobra Venom Factor 与 前 の値 に対 す る 百 分 率 に て 表 示 し た. ラ ッ トC3の 測 定:抗 (Cappel社)を SRID法 ラ ッ トC3ヤ ギ抗 体 用 い て 自家 製 プ レー トを 作 製 し に て 測 定 した. C3もCVF, 与 前 値 に 対 す る百 分 率 で 表 示 した. CRAの 測 定: ASS rabbitsに と同様 に行 い, CVF, おけ る測定 法 与前値 に対 す る百 図2 CSS ratsの 作 製及 び 腎生 検 分 率 で 表 示 した. 正 常 ラ ッ ト4匹 にCVF50Uを 毎週腹 腔へ 注 射 し, 1週 後 よ り毎 週 腎 生 検,採 CVFの 結 1. ASS 1) 血 を施 行 し, 腎及 び 補 体 に 対 す る影 響 を検 討 した. rabbits ASS ASS 果 rabbitsに nephritis作 ギIgG,ウ サ ギC3の お け るICの 製 後,腎 消長 局 所 のBSA,ウ 沈 着 をscoreに サ て表示 し た.(表2),(図3) 図3 (1) BSA 与 群 で は, BSAは 0.45(mean±1SD), 後0.45±0.55, 1週 後0.66±0.57, 3週 後0で 2週 後1.33±0.96, 1.15で あ り, 1週 後 のBSA沈 2週 1週 後 3週 後1.00± 着 にの み両者 に 有 意 差(p<0.05)が 認 め ら れ た.即 与 群 の 方 が 与 群 に 比 べBSAの ち 沈着 の と も 消 失).で あ っ た. 1.10±0.87, 2週 3週 消長 与 群 で は, 1週 後1.44±1.01, 後0(5匹 2週 と も 消 失)で 沈 着 と 同 様, 後 あ っ 与 群 の 方 がCVF 投 与 群 に 比 べ て ウ サ ギIgGの 沈着 の消失 が早い な か っ た. 0.45, 投 与 前1.60±0.55, 後0.20±0.45, 3週 た. BSAの C3 与 群 の ウ サ ギIgGは 後0.66±0.58, 0(5匹 (3) ウ サ ギIgG 1週 お け るICの 傾 向 が 認 め られ た が い ず れ も有 意 差 は 認 め られ 消 失 が 早 い 傾 向 が 認 め られ た.(図4) (2) rabbitsに 投 与 前1.60±0.55, あ っ た. 与 群 で は,投 与 前2.00±0.71, 1.80±0.69, ASS 投 与 前1.80± 後 3週 与 群 の ウ サ ギC3は,投 1週 後1.66±0.58, 後1.00で あ っ た. 2週 与 前2.20± 後1.00±0.71, 与群 では 投 与前
4 1054 馬 図4 ASS rabbitsに 越 由 仲 おけ る腎糸球 体 の蛍光 所 見 A: 与 後BSA(1週 D: 与 後BSA(1週 B: 与 後IgG(1週 E: 与 後IgG(1週 C: 与 後C3(1週 F: 与 後C3(1週 与 群 に お い て 腎 糸 球 体 へ の 沈 着 の 消 失 遅 延 が 認 め ら れ る.
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9 1) Miller GW and Nussenzweig V: A new complement function. Solubilization of antigen-antibody aggregates. Proc Natl Acad Sci USA (1975) 72, ) Shifferli JA, Bartolotti SR and Peters DK: Inhibition of immune precipitation by complement. Clin Exp Immunol (1980) 42, ) Agnello V: Complement deficiency states. Medicine (1978) 57, ) Mayer MM: Complement and Complement Fixation in Experimental Immunochemistry, 2nd Ed by Kabat, Mayer, Springfield III CC Thomas (1961) pp ) Amano T, Aibara Y, Kuwajima N, Suzuki S and Ota Z: A simplified quantification method of complex-release activity using peroxidase as immune complex antigen. Acta Med Okayama (1983) 37, ) Arisz I, Noble B, Milgrom M, Brentjens JR and Andres GA: Experimental chronic serum sickness in rats. A model of immune complex glomerulonephritis Int Arch Allergy Appl Immunol (1979) 60, and systemic immune complex deposition. 8) Takahashi M, Takahashi S and Hirose S: Solubilization of antigen-antibody complexes: A new function of complement as a regulator of immune reactions. Prog Allerg (1980) 27, ) Bartolloti SR and Peters DK: Delayed removal of renal-bound antigen in decomplemented rabbits with acute serum sickness. Clin Exp Immunol (1978) 32,
10 Complement-mediated solubilization of immune complexes in vivo Yoshinaka UMAKOSHI Third Department of Internal Medicine, Okayama University Medical School, Okayama (Director: 700, Japan Prof. Z. Ota) In order to examine the ability of complement to solubilize immune complexes (IC) in vivo, acute or chronic serum sickness nephritis was induced in 10 rabbits and 11 rats with by injection of BSA as an antigen (Ag). The animals were divided into two groups. The first received 500U or 50U/week cobra venom factor (CVF) administered intraperitoneally to reduce serum complement levels and complement-mediated solubilization of immune complex es. On the other hand, the second group received 2ml or 0.2ml/week turpentine oil (TO) injected interamuscularly to elevate serum complement levels and complement-mediated solubilization of immune complex. Renal biopsies were performed weekly with each animal, and these specimens were stained for antigen (BSA), antibody (IgG) and complement (C3) by immunofluorescence techniques. Results showed a significant difference between TO-, and CVF-treated animals with respect to the decline of IC in kidney lesions. No detectable IC (especially BSA) remained in the renal glomeruli of TO-treated nephritic animals, indicating that complement might act in vivo to solubilize IC deposited in tissues.
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