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.
Immunopathology Laboratory, Butantan Institute, Av. Dr. Vital Brazil, 1500, Butantan, Sao Paulo 05503-900, SP, Brazil. clissapb@butantan.gov.br
P B
PB
Clissa
M
M
Lopes-Ferreira
M S
MS
Della-Casa
S H P
SH
Farsky
A M
AM
Moura-da-Silva
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