Reddy D N, Chitko-McKown C G, Reddy P G, Minocha H C, Blecha F
Department of Anatomy and Physiology, Kansas State University, Manhattan 66506.
Vet Immunol Immunopathol. 1993 Feb;36(1):17-29. doi: 10.1016/0165-2427(93)90003-m.
We describe the preparation of monoclonal antibodies (Mabs) directed against recombinant bovine interleukin-1 beta (rBoIL-1 beta). These anti-IL-1 beta Mabs were designated SA10, SA12, SA13, SA15, and SA22, and were characterized on the basis of their epitope specificity and cross-reactivity with homologous and heterologous cytokines in enzyme-linked immunosorbent assays and immunoblot analyses. Additionally, the ability of these Mabs to neutralize IL-1 beta was tested in thymocyte costimulation assays. The ELISA titers of all Mabs ranged from 9.4 x 10(6) to 1 x 10(7). Data indicate that Mabs SA10, SA12, SA15, and SA22 neutralized both bovine macrophage-derived IL-1 (1:4) and rBoIL-1 beta (1 ng ml-1). All the Mabs against rBoIL-1 beta (SA10, SA12, SA13, SA15, SA22) were specific and did not cross-react with other cytokines tested, except recombinant human IL-1 beta (rHuIL-1 beta). This finding suggests that these Mabs recognize epitopes common to human and bovine IL-1 molecules. Competition experiments suggested that Mab SA22 recognized a different epitope and Mabs SA10, SA12, SA13, and SA15 recognized the same epitope on the rBoIL-1 beta molecule. These observations suggest that these Mabs could be useful reagents for developing immunoassays to measure bovine IL-1 beta from biological fluids and to study the immunoregulatory role of IL-1 in the bovine immune system.
我们描述了针对重组牛白细胞介素-1β（rBoIL-1β）的单克隆抗体（Mab）的制备。这些抗IL-1β单克隆抗体被命名为SA10、SA12、SA13、SA15和SA22，并通过酶联免疫吸附测定和免疫印迹分析，根据其表位特异性以及与同源和异源细胞因子的交叉反应性进行了表征。此外，在胸腺细胞共刺激试验中测试了这些单克隆抗体中和IL-1β的能力。所有单克隆抗体的酶联免疫吸附测定效价范围为9.4×10⁶至1×10⁷。数据表明，单克隆抗体SA10、SA12、SA15和SA22可中和牛巨噬细胞衍生的IL-1（1:4）和rBoIL-1β（1 ng/ml）。所有针对rBoIL-1β的单克隆抗体（SA10、SA12、SA13、SA15、SA22）均具有特异性，除重组人IL-1β（rHuIL-1β）外，不与其他测试的细胞因子发生交叉反应。这一发现表明，这些单克隆抗体识别的是人和牛IL-1分子共有的表位。竞争实验表明，单克隆抗体SA22识别不同的表位，而单克隆抗体SA10、SA12、SA13和SA15识别rBoIL-1β分子上相同的表位。这些观察结果表明，这些单克隆抗体可能是开发免疫测定以测量生物体液中牛IL-1β以及研究IL-1在牛免疫系统中免疫调节作用的有用试剂。
