title: A serine protease isolated from the bristles of the Amazonic caterpillar, Premolis semirufa, is a potent complement system activator.
abstract: BACKGROUND
  The caterpillar of the moth Premolis semirufa, commonly named pararama, is found in the Brazilian Amazon region. Accidental contact with the caterpillar bristles causes an intense itching sensation, followed by symptoms of an acute inflammation, which last for three to seven days after the first incident. After multiple accidents a chronic inflammatory reaction, called "Pararamose", characterized by articular synovial membrane thickening with joint deformities common to chronic synovitis, frequently occurs. Although complement mediated inflammation may aid the host defense, inappropriate or excessive activation of the complement system and generation of anaphylatoxins can lead to inflammatory disorder and pathologies. The aim of the present study was to evaluate, in vitro, whether the Premolis semirufa's bristles extract could interfere with the human complement system.
  
  
  RESULTS
  The bristles extract was able to inhibit the haemolytic activity of the alternative pathway, as well as the activation of the lectin pathway, but had no effect on the classical pathway, and this inhibition seemed to be caused by activation and consumption of complement components. The extract induced the production of significant amounts of all three anaphylatoxins, C3a, C4a and C5a, promoted direct cleavage of C3, C4 and C5 and induced a significant generation of terminal complement complexes in normal human serum. By using molecular exclusion chromatography, a serine protease of 82 kDa, which activates complement, was isolated from P. semirufa bristles extract. The protease, named here as Ps82, reduced the haemolytic activity of the alternative and classical pathways and inhibited the lectin pathway. In addition, Ps82 induced the cleavage of C3, C4 and C5 and the generation of C3a and C4a in normal human serum and it was capable to cleave human purified C5 and generate C5a. The use of Phenanthroline, metalloprotease inhibitor, in the reactions did not significantly interfere with the activity of the Ps82, whereas the presence of PMSF, serine protease inhibitor, totally blocked the activity.
  
  
  CONCLUSIONS
  These data show that a serine protease present in the Premolis semirufa's bristles extract has the ability to activate the complement system, which may contribute to the inflammatory process presented in humans after envenomation.
authors.affiliation: Immunochemistry Laboratory, Butantan Institute, São Paulo, SP, Brazil.
authors.forename: Isadora Maria
authors.initials: IM
authors.lastname: Villas Boas
authors.affiliation: Immunochemistry Laboratory, Butantan Institute, São Paulo, SP, Brazil.
authors.forename: Giselle
authors.initials: G
authors.lastname: Pidde-Queiroz
authors.affiliation: Immunochemistry Laboratory, Butantan Institute, São Paulo, SP, Brazil.
authors.forename: Fabio Carlos
authors.initials: FC
authors.lastname: Magnoli
authors.affiliation: Immunochemistry Laboratory, Butantan Institute, São Paulo, SP, Brazil.
authors.forename: Rute M
authors.initials: RM
authors.lastname: Gonçalves-de-Andrade
authors.affiliation: Institute of Molecular and Experimental Medicine, School of Medicine, Cardiff University, Cardiff, United Kingdom.
authors.forename: Carmen W
authors.initials: CW
authors.lastname: van den Berg
authors.affiliation: Immunochemistry Laboratory, Butantan Institute, São Paulo, SP, Brazil.
authors.forename: Denise V
authors.initials: DV
authors.lastname: Tambourgi
mesh_terms: D000706:Anaphylatoxins / Q000737:chemistry / Q000302:isolation & purification; D000818:Animals; D003167:Complement Activation / Q000187:drug effects*; D015938:Complement Membrane Attack Complex / Q000737:chemistry; D004912:Erythrocytes / Q000187:drug effects; D006801:Humans; D019476:Insect Proteins / Q000302:isolation & purification / Q000494:pharmacology*; D009036:Moths / Q000201:enzymology*; D059748:Proteolysis; D057057:Serine Proteases / Q000302:isolation & purification / Q000494:pharmacology*
chemical_list: D000706:Anaphylatoxins; D015938:Complement Membrane Attack Complex; D019476:Insect Proteins; D057057:Serine Proteases