Soluble type-I interleukin-1 receptor blocks chicken IL-1 activity
Soluble type-I interleukin-1 receptor blocks chicken IL-1 activity
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DOI：
10.1016/s0145-305x(00)00064-1
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发表时间：
2001-05-01
影响因子：
2.9
通讯作者：
Peng, RK
中科院分区：
文献类型：
--
作者：
Klasing, KC;Peng, RK
将鸡I型白细胞介素1（IL-1）受体（可溶性IL-1 R（I）; sIL-1 R（I））的配体结合域克隆到毕赤酵母表达系统中，并将所得sIL-1 R（1）结合蛋白用于制备兔抗血清（抗IL-1 R（1））。进行了两个实验以确定sIL-1 R（1）或抗IL-1 R（1）在来自经脂多糖刺激的HD 11鸡巨噬细胞的条件培养基（CM）中阻断IL-1生物活性（胸腺细胞共刺激）的能力。在第一个实验中，CM与未纯化的sIL-1 R（1）预孵育，其胸腺细胞共刺激活性显著降低57%。用分子排阻过滤法或60%硫酸铵沉淀法从毕赤酵母表达系统分泌的其它蛋白中进一步纯化sIL-1 R（1），并不影响其中和IL-1活性的能力。这些部分纯化的sIL-1 R（1）制剂分别使CM中的胸腺细胞共刺激活性降低70.7%和77.3%。在第二个实验中，用抗sIL-1 R（1）的抗血清预孵育胸腺细胞，相对于未接受抗体的对照胸腺细胞培养物，CM中的IL-1活性降低了70%，相对于用免疫前血清孵育的胸腺细胞培养物，降低了59%。推测抗sIL-1 R（1）通过阻断IL-1与胸腺细胞上的I型受体结合而降低CM中IL-1的生物活性。因此，LPS刺激的HD 11巨噬细胞释放的IL-1样活性的30%可能是由于至少一种其他细胞因子。我们的数据是一致的I型受体是鸡胸腺细胞上的主要IL-1受体，能够提供增殖的信号。(C)2001爱思唯尔科技有限公司版权所有。
The ligand-binding domain of the chicken type-I interleukin-1 (IL-1) receptor (soluble IL-1R(I); sIL-1R(I)) was cloned into a Pichia pastoris expression system and the resulting sIL-1R(1) binding protein was used to produce antisera in rabbits (anti-IL-1R(1)). Two experiments were conducted to determine the capacity of sIL-1R(1) or anti-IL-1R(1) to block the IL-1 bioactivity (thymocyte co-stimulation) in conditioned media (CM) from HD11 chicken macrophages stimulated with lipopolysaccharide. In the first experiment, pre-incubation of CM with unpurified sIL-1R(1) significantly decreased its thymocyte co-stimulation activity by 57%. Further purification of sIL-1R(1) from other proteins secreted ol shed from P. pastoris expression system by size exclusion filtration or ammonium sulfate (60%) precipitation did not influence its capacity to neutralize IL-1 bioactivity, These partially purified sIL-1R(1) preparations significantly decreased thymocyte co-stimulation activity in CM by 70.7 and 77.3%, respectively. In the second experiment, pre-incubation of thymocytes: with antisera against the sIL-1R(1) decreased IL-1 activity in CM by 70% relative to control thymocyte cultures that received no antibody and by 59% relative to thymocyte cultures incubated with pre-immune sera. Presumably anti-sIL-1R(1) diminished the IL-1 bioactivity in CM by blocking IL-1 binding to its type-I receptor on thymocytes. Thus, 30% of the IL-1-like activity released by LPS-stimulated HD11 macrophages is probably due to at least one other cytokine. Our data are consistent with the type-I receptor being the primary IL-1 receptor on chicken thymocytes that is capable of providing a signal for proliferation. (C) 2001 Elsevier Science Ltd. All rights reserved.