CD44 and hyaluronic acid regulate in vivo iNOS expression and metalloproteinase activity in murine air-pouch inflammation.
OBJECTIVE
To evaluate the effects of anti-CD44 IM7.8.1 antibody, HMW-HA and LMW-HA on leukocyte migration and adhesion, and the induction of proinflammatory mediators, in mouse air-pouch inflammation induced by zymosan.


METHODS
Leukocytes were obtained from zymosan-air pouches after the intra-pouch injection of anti-CD44 IM7.8.1, isotype control, HMW-HA, LMW-HA or PBS. TNF-alpha, IL-1beta and iNOS mRNA were estimated in leukocytes by semi-quantitative RT-PCR. Matrix metalloproteinases (MMPs) from exudates were evaluated by zymography and Western Blot. Adhesion and migration of leukocytes were evaluated in HA-coated plates and Boyden chambers respectively.


RESULTS
IM7.8.1 decreased iNOS mRNA levels and the activity of both MMP-9 and MMP-2 eight h after injection into zymosan air pouch while IM7.8.1, HMW-HA and LMW-HA had no effect on IL1-beta or TNF-alpha mRNA levels. Leukocytes from air pouch adhered to and migrated in vitro against both HMW-HA and LMW-HA. LMW-HA increased the number of leukocytes in the air pouch and iNOS mRNA levels as compared to PBS injection. In contrast, HMW-HA decreased leukocyte count and reduced iNOS mRNA levels. Paradoxically, the activity of both MMP-9 and MMP-2 was increased by HMW-HA and decreased by LMW-HA.


CONCLUSIONS
Both CD44 and HA can modulate leukocyte migration and induction of proinflammatory mediators in mouse zymosan air pouch inflammation. IM7.8.1 had consistent anti-inflammatory effects, reducing iNOS, MMP-9 and MMP-2. HMW-HA and LMW-HA were able to modulate both the induction of proinflammatory mediators and leukocyte count in the air pouch.
Cabrera
P V
PV
Cátedra de Inmunología-IDEHU, Facultad de Farmacia y Bioquímica, Universidad Nacional de Buenos Aires (UBA)-CONICET, Junín 956, piso 4, 1113, Buenos Aires, Argentina.
Blanco
G
G
Alaniz
L
L
Greczanik
S
S
Garcia
M
M
Alvarez
E
E
Hajos
S E
SE
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D015153:Blotting, Western; D002448:Cell Adhesion; D002465:Cell Movement; D002633:Chemotaxis; D016207:Cytokines / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D006495:Heparin, Low-Molecular-Weight / Q000378:metabolism; D018960:Hyaluronan Receptors / Q000096:biosynthesis* / Q000737:chemistry; D006820:Hyaluronic Acid / Q000096:biosynthesis* / Q000737:chemistry; D007249:Inflammation; D007375:Interleukin-1 / Q000378:metabolism; D007962:Leukocytes / Q000166:cytology / Q000378:metabolism; D020782:Matrix Metalloproteinases / Q000378:metabolism*; D051379:Mice; D019001:Nitric Oxide Synthase / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II; D016133:Polymerase Chain Reaction; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D000911:Antibodies, Monoclonal; D016207:Cytokines; D017931:DNA Primers; D006495:Heparin, Low-Molecular-Weight; D018960:Hyaluronan Receptors; D007375:Interleukin-1; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D012313:RNA; D006820:Hyaluronic Acid; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse; D020782:Matrix Metalloproteinases