Leucocyte recruitment induced by type II phospholipases A(2) into the rat pleural cavity

Bothropstoxin-I (BthTX-I) and bothropstoxin-II (BthTX-II) are Lys-49 and As p-49 phospholipases A(2) (PLA(2)s), respectively, isolated from Bothrops ja raracussu venom. Piratoxin-I (PrTX-I) is a Lys-49 PLA? isolated from Bothro ps pirajai venom. In this study, the ability of BthTX-I, BthTX-II and PrTX- I to recruit leucocytes into the rat pleural cavity and potential mechanism s underlying this effect were investigated. Intrapleural injection of eithe r BthTX-I or PrTX-I (10-100 mu g/cavity each) caused a significant leucocyt e infiltration at 12 h after injection. The maximal cell migration was obse rved with the dose of 30 mu g/cavity (14.9 +/- 15.5 and 17.6 +/- 1.6 x 10(6 ) cells/cavity, respectively). Leucocyte counts consisted mainly of mononuc lear cells, but significant amounts of neutrophils and eosinophils were als o observed. Intrapleural injection of BthTX-II (10-100 mu g/cavity) caused a marked leucocyte infiltration at 6 and 12 h after injection. The maximal response was observed with the dose of 100 mu g/cavity (57.3 +/- 3.4 x 10(6 ) cells/cavity, 6 h). The leucocyte counts were mainly composed of neutroph ils and mononuclear cells. The treatment of either BthTX-I (30 mu g/cavity, 12 h) or BthTX-II (30 mu g/cavity, 6 h) with the PLA(2) inhibitor p-bromop henacyl bromide (p-BPB) had no effect on the total and differential leucocy te counts induced by these proteins. The same treatment partially reduced t he PrTX-I-induced pleural leucocyte infiltration. In rats depleted of the h istamine and 5-hydroxytryptamine (5-HT) stores by chronic treatment with co mpound 48/80, the total leucocyte counts in response to BthTX-I, BthTX-II a nd PrTX-I was not significantly affected compared to control animals. In ad dition, BthTX-II BthTX-II and PrTX-I (100 mu g/ml each) significantly degra nulated pleural mast cells in vitro leading to the release of [C-14]5-hydro xytryptamine ([C-14]S-HT). p-BPB and heparin (50 IU/ml) significantly reduc ed the [C-14]5-HT release induced by these PLA(2)s. Our results demonstrate that BthTX-I, BthTX-II and PrTX-I recruit leucocyte into the pleural cavit y of the rat by mechanisms unrelated to enzymatic activity and pleural mast cell degranulation. (C) 2000 Elsevier Science Ltd. All rights reserved.