Minnerly J C, Baganoff M P, Deppeler C L, Keller B T, Rapp S R, Widomski D L, Fretland D J, Bolanowski M A
Searle Research & Development, St. Louis, Missouri 63198, USA.
Inflammation. 1995 Jun;19(3):313-31. doi: 10.1007/BF01534390.
To establish a direct link between IL-8 and inflammation in vivo, we first isolated the gene encoding rhesus macaque IL-8. The open reading frame directs the translation of a 101 amino acid (aa) precursor, which is 94% identical to human IL-8. Rhesus IL-8 was expressed in bacteria and purified to homogeneity with ion-exchange chromatography. Pure rhesus IL-8 was biologically active as measured by its ability to bind specifically to either rhesus (Kd = 0.5 nM) or human (Kd = 2 nM) IL-8 receptors and to promote in vitro chemotaxis of rhesus (EC50 = 2 nM) or human neutrophils (EC50 = 4 nM). Moreover, a mouse monoclonal antibody, DM/C7, which neutralizes human IL-8 activity, also recognized and neutralized (IC50 = 0.5-3.0 microgram/ml) rhesus IL-8 in vitro. Systemic administration of DM/C7 completely inhibited the dermal inflammation of rhesus ears induced by the external application of phorbol myristoyl acetate. These observations reveal that rhesus IL-8 is structurally and functionally similar to human IL-8 and suggests that IL-8 plays a prominent role in a primate model of inflammation.
为了在体内建立白细胞介素-8（IL-8）与炎症之间的直接联系，我们首先分离了编码恒河猴IL-8的基因。开放阅读框指导翻译出一个101个氨基酸（aa）的前体，它与人类IL-8有94%的同源性。恒河猴IL-8在细菌中表达，并通过离子交换色谱法纯化至同质。通过其与恒河猴（解离常数Kd = 0.5 nM）或人类（Kd = 2 nM）IL-8受体特异性结合的能力以及促进恒河猴（半数有效浓度EC50 = 2 nM）或人类中性粒细胞体外趋化性（EC50 = 4 nM）来测定，纯化后的恒河猴IL-8具有生物活性。此外，一种能中和人类IL-8活性的小鼠单克隆抗体DM/C7，在体外也能识别并中和（半数抑制浓度IC50 = 0.5 - 3.0微克/毫升）恒河猴IL-8。全身给予DM/C7可完全抑制外用佛波醇肉豆蔻酸酯诱导的恒河猴耳部皮肤炎症。这些观察结果表明，恒河猴IL-8在结构和功能上与人类IL-8相似，并提示IL-8在灵长类动物炎症模型中起重要作用。
