Importance of jararhagin disintegrin-like and cysteine-rich domains in the early events of local inflammatory response.
Jararhagin is a multi-domain SVMP from Bothrops jararaca venom comprising catalytic, disintegrin-like and cysteine-rich domains, which cause a local reaction manifested by hemorrhage, edema, cytokine release and inflammatory cell recruitment. In this study, the importance of disintegrin-like/cysteine-rich domains of jararhagin was addressed by analyzing the effects of jararhagin-C, which lacks the catalytic domain, in induction of leukocyte rolling and release of pro-inflammatory cytokines. Jararhagin-C was isolated from B. jararaca venom conserving the same ability of complete jararhagin molecule in inhibiting collagen-induced platelet-aggregation. Treatment of trans-illuminated cremaster muscle in vivo with jararhagin-C increased number of rolling leukocytes (approximately 250%) in post-capillary venules in all periods analyzed, without interfering with microvasculature haemodynamic, like vessel diameter, the erythrocyte speed or the blood flow rate. The release of pro-inflammatory cytokines TNF-alpha, IL-1beta and IL-6 was significantly enhanced in the local of jararhagin-C injection, showing the maximum levels in periods between 2 and 4 h after treatment. Besides the action of jararhagin-C, the presence of the inactivated catalytic domain in o-phenanthrolin-treated jararhagin was related to a higher increase in the number of rolling leukocytes. Moreover, the levels of IL-6 and IL-1beta induced by catalytically active jararhagin were higher than those induced by jararhagin-C. In conclusion, our findings suggest that the disintegrin-like/cysteine-rich domains of jararhagin are sufficient to locally activate the early events of an acute inflammatory response as leukocyte rolling and pro-inflammatory cytokine release and this action may add to the effect of catalysis, which enhances the primary cell activation.
Clissa
P B
PB
Immunopathology Laboratory, Butantan Institute, Av. Dr. Vital Brazil, 1500, Butantan, Sao Paulo 05503-900, SP, Brazil. clissapb@butantan.gov.br
Lopes-Ferreira
M
M
Della-Casa
M S
MS
Farsky
S H P
SH
Moura-da-Silva
A M
AM
D000818:Animals; D017837:Bothrops / Q000378:metabolism; D020134:Catalytic Domain; D003435:Crotalid Venoms / Q000737:chemistry* / Q000633:toxicity*; D003545:Cysteine / Q000737:chemistry*; D016207:Cytokines; D019483:Disintegrins / Q000737:chemistry*; D004730:Endothelium, Vascular / Q000378:metabolism; D007249:Inflammation / Q000139:chemically induced* / Q000473:pathology; D015850:Interleukin-6 / Q000378:metabolism; D036904:Leukocyte Rolling / Q000187:drug effects; D008666:Metalloendopeptidases / Q000737:chemistry* / Q000633:toxicity*; D051379:Mice; D018482:Muscle, Skeletal / Q000098:blood supply / Q000166:cytology; D010975:Platelet Aggregation Inhibitors; D013997:Time Factors; D014699:Venules; D000097146:Bothrops jararaca Venom
D003435:Crotalid Venoms; D003545:Cysteine; D016207:Cytokines; D019483:Disintegrins; D015850:Interleukin-6; D008666:Metalloendopeptidases; D010975:Platelet Aggregation Inhibitors; D000097146:Bothrops jararaca Venom