Immunoregulation of hepatic fibrosis in murine schistosomiasis japonica.
Immunoregulation of hepatic fibrosis in murine schistosomiasis japonica.
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小鼠日本血吸虫病肝纤维化的免疫调节。
DOI：
10.1093/infdis/159.4.798
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发表时间：
1989
期刊：
影响因子：
--
通讯作者：
Kresina,TF
中科院分区：
文献类型：
--
作者：
Olds,GR;Kresina,TF
方法为检测免疫抑制性T细胞或免疫球蛋白是否可以调节日本血吸虫感染时的肝纤维化，制备未感染的C57BL/6小鼠或感染10周或30周的小鼠的血清池和脾淋巴细胞。从这些动物获得的细胞或血清样本被转移到急性感染的受体小鼠中。根据为肉芽肿性炎症的最大体内调节而建立的方案施用血清或细胞[6, 7]：在感染第4周和第5周时转移0.5 mL血清或4 x 10'细胞。 6周时处死受体动物，取出肝脏切片用于测定平均肉芽肿面积[9]：器官的其余部分用于测定胶原代谢的各种参数[8]。这些包括通过羟脯氨酸含量测定的肝胶原含量[8]，通过将3H脯氨酸掺入3H羟脯氨酸来估计胶原生物合成[8]，以及通过将3H脯氨酸掺入胃蛋白酶可提取的不可透析盐沉淀胶原中来估计细胞外胶原合成[8]。此外，新合成的胶原蛋白的同种型通过胃蛋白酶提取、差别盐沉淀以及 A-5M 琼脂糖和羧甲基纤维素色谱法测定[10]。
MethodsTo examine whether immunosuppressive T cells or immunoglobulin could modulate hepatic fibrosis in S. japoni-cum infection, serum pools and splenic lymphoid cells were prepared from uninfected C57BL/6 mice or mice infected for 10 or 30 w. Cells or serum samples obtained from these animals were transferred into acutely infected recipient mice. Sera or cells were administered according to protocols established for maximal in vivo modulation of granuloma-tous inflammation [6, 7]: 0.5 mL of serum or 4 x 10'cells were transferred at 4 and 5 w of infection. Recipient animals were killed at 6 w and a section of the liver was removed for the determination of average granuloma area [9]: the rest of the organ was used to determine various parameters of collagen metabolism [8]. These included he-patic collagen content determined by hydroxyproline con-tent [8], collagen biosynthesis estimated by the incorpo-ration of 3H proline into 3H hydroxyproline [8], and extracellular collagen synthesis by the incorporation of 3H proline into pepsin extractable nondialyzable salt precipita-ble collagen [8]. Also, the isotype of newly synthesized collagen was determined by pepsin extraction followed by differential salt precipitation and A-5M agarose and carboxymethyl cellulose chromatography [10].