Recombinant Human Hyaluronidase PH20 Does Not Stimulate an Acute Inflammatory Response and Inhibits Lipopolysaccharide-Induced Neutrophil Recruitment in the Air Pouch Model of Inflammation
Recombinant Human Hyaluronidase PH20 Does Not Stimulate an Acute Inflammatory Response and Inhibits Lipopolysaccharide-Induced Neutrophil Recruitment in the Air Pouch Model of Inflammation
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DOI：
10.4049/jimmunol.1303060
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发表时间：
2014-06-01
影响因子：
4.4
通讯作者：
Shepard, H. Michael
中科院分区：
文献类型：
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作者：
Huang, Zhongdong;Zhao, Chunmei;Shepard, H. Michael
透明质酸酶(Hyal)和低分子量。透明质酸(LMW HA)片段被广泛报道能刺激先天免疫反应。然而，所使用的大多数透明质酸酶是从动物组织(例如牛睾丸组织[BTH])中提纯的，并且含有内毒素和其他无关的蛋白质。我们在啮齿动物气囊炎症模型中测试了高纯度的重组人Hyal(RHuPH20)和不含内毒素的HA片段，其M-r从5,000到1,500,000，以确定它们刺激先天性免疫反应的可能性。外源性LMW HA片段(平均MR为200,000)既不能诱导细胞因子/趋化因子的产生，也不能诱导中性粒细胞渗入气囊。用LPS或BTH刺激气囊可刺激细胞因子和趋化因子的产生，但rHuPH20不能，提示PH20和原位LMW HA片段的产生都不能刺激细胞因子和趋化因子的产生。脂多糖和BTH还可诱导中性粒细胞渗入气囊，而rHuPH20处理组未见此现象。去除内毒素的BTH的促炎活性明显降低，提示rHuPH20和BTH的炎症反应差异可能是由于BTH中的内毒素污染所致。当rHuPH20与内毒素共同作用时，细胞因子和趋化因子的诱生作用与单纯内毒素作用相同，但中性粒细胞的浸润受到抑制，可能是通过阻断HA-CD44相互作用而实现的。我们的结果表明，在气囊模型中，rHuPH20及其直接生成的HA分解代谢物都没有炎症特性，相反，rHuPH20可以抑制某些方面的炎症，如中性粒细胞渗入气囊。
Hyaluronidase (Hyal) and low m.w. hyaluronan (LMW HA) fragments have been widely reported to stimulate the innate immune response. However, most hyaluronidases used were purified from animal tissues (e.g., bovine testis Hyal [BTH]), and contain endotoxin and other unrelated proteins. We tested a highly purified recombinant human Hyal (rHuPH20) and endotoxin-free HA fragments from M-r 5,000 to 1,500,000 in the rodent air pouch model of inflammation to determine their potential for stimulation of the innate immune response. Exogenous LMW HA fragments (average Mr 200,000) failed to induce either cytokine/chemokine production or neutrophil infiltration into the air pouch. Challenging the air pouch with LPS or BTH stimulated production of cytokines and chemokines but rHuPH20 did not, suggesting that neither PH20 nor generation of LMW HA fragments in situ stimulates cytokine and chemokine production. LPS and BTH also induced neutrophil infiltration into the air pouch, which was not observed with rHuPH20 treatment. Endotoxin-depleted BTH had much reduced proinflammatory activity, suggesting that the difference in inflammatory responses between rHuPH20 and BTH is likely due to endotoxin contaminants in BTH. When rHuPH20 was dosed with LPS, the induction of cytokines and chemokines was the same as LPS alone, but neutrophil infiltration was inhibited, likely by interrupting HA-CD44 interaction. Our results indicate that neither rHuPH20 nor its directly generated HA catabolites have inflammatory properties in the air pouch model, and rHuPH20 can instead inhibit some aspects of inflammation, such as neutrophil infiltration into the air pouch.