Are integrins involved in the aggregatory and phagocytic behaviour of fish haemostatic cells?
The involvement of a putative integrin-like fibrinogen receptor in the aggregatory and phagocytic behaviour of thrombocytes (platelet equivalents of fish) from the rainbow trout Oncorhynchus mykiss was studied. Aggregation of trout thrombocytes was induced by the thromboxane mimetic U-46619 in the presence of trout fibrinogen. Thrombocyte aggregation was inhibited by the tetrapeptide RGDS, but not by RGES or fibrinogen binding inhibitor peptide (HHLGGAKQAGDV). A range of monoclonal antibodies against the human platelet integrin alphaIIbbeta3 (anti-CD41a, anti-beta3 and LK7r) showed no reactivity with trout thrombocytes. Subsequently, a panel of monoclonal antibodies was raised against thrombocyte membrane preparations in an attempt to obtain an antibody against the putative integrin fibrinogen receptor. Of these monoclonal antibodies, four were found to inhibit thrombocyte aggregation, namely 12G2, 30D8, 32F8 and 32H10. The antibody 32H10 was shown significantly to inhibit the attachment of thrombocytes to immobilised trout fibrinogen, suggesting that it and the other antibodies recognise the putative fibrinogen receptor on trout thrombocytes. FITC-labelled Bacillus cereus were employed as test particles to prove that thrombocytes internalise bacteria via an active process and not simply by passive sequestration into the open canalicular system. Preincubation of bacteria with trout fibrinogen resulted in a significant increase in the number of thrombocytes exhibiting phagocytosis. This enhancement of phagocytosis by preincubation of B. cereus with trout fibrinogen could be inhibited by the tetrapeptide RGDS, but not by RGES, hence implicating the putative fibrinogen receptor in the internalisation of microorganisms. The relevance of these findings to the possible existence of an integrin-like receptor on trout thrombocytes is discussed.
Hill
D J
DJ
School of Biological Sciences, University of Wales, Swansea, Singleton Park, Swansea SA2 8PP, Wales, UK.
Rowley
A F
AF
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001777:Blood Coagulation; D001792:Blood Platelets / Q000502:physiology* / Q000648:ultrastructure; D005340:Fibrinogen / Q000378:metabolism; D006487:Hemostasis; D016023:Integrins / Q000502:physiology*; D017686:Oncorhynchus mykiss / Q000502:physiology*; D010587:Phagocytosis; D010974:Platelet Aggregation; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000502:physiology
D000911:Antibodies, Monoclonal; D016023:Integrins; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D005340:Fibrinogen