Recombinant human hyaluronidase PH20 does not stimulate an acute inflammatory response and inhibits lipopolysaccharide-induced neutrophil recruitment in the air pouch model of inflammation.
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 M(r) 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.
Halozyme Therapeutics, San Diego, CA 92121 jhuang@halozyme.com.
Zhongdong
Z
Huang
Halozyme Therapeutics, San Diego, CA 92121.
Chunmei
C
Zhao
Halozyme Therapeutics, San Diego, CA 92121.
Yanling
Y
Chen
Halozyme Therapeutics, San Diego, CA 92121.
Jessica A
JA
Cowell
Halozyme Therapeutics, San Diego, CA 92121.
Ge
G
Wei
Halozyme Therapeutics, San Diego, CA 92121.
Anne
A
Kultti
Halozyme Therapeutics, San Diego, CA 92121.
Lei
L
Huang
Halozyme Therapeutics, San Diego, CA 92121.
Curtis B
CB
Thompson
Halozyme Therapeutics, San Diego, CA 92121.
Sanna
S
Rosengren
Halozyme Therapeutics, San Diego, CA 92121.
Gregory I
GI
Frost
Halozyme Therapeutics, San Diego, CA 92121.
H Michael
HM
Shepard
D000208:Acute Disease; D000818:Animals; D000951:Antigens, Neoplasm / Q000276:immunology / Q000494:pharmacology*; D002417:Cattle; D002460:Cell Line; D016207:Cytokines / Q000276:immunology; D051548:Histone Acetyltransferases / Q000276:immunology / Q000494:pharmacology*; D006801:Humans; D006820:Hyaluronic Acid / Q000276:immunology*; D006821:Hyaluronoglucosaminidase / Q000276:immunology / Q000494:pharmacology*; D007249:Inflammation / Q000139:chemically induced / Q000276:immunology / Q000473:pathology; D008070:Lipopolysaccharides / Q000633:toxicity*; D008297:Male; D051379:Mice; D020556:Neutrophil Infiltration / Q000187:drug effects* / Q000276:immunology; D009504:Neutrophils / Q000276:immunology* / Q000473:pathology; D011994:Recombinant Proteins / Q000276:immunology / Q000494:pharmacology
D000951:Antigens, Neoplasm; D016207:Cytokines; D008070:Lipopolysaccharides; D011994:Recombinant Proteins; D006820:Hyaluronic Acid; D051548:Histone Acetyltransferases; C115866:OGA protein, human; D006821:Hyaluronoglucosaminidase