The production and characterisation of monoclonal antibodies to Lucilia cuprina larval antigens.
A panel of murine monoclonal antibodies was produced against three Lucilia cuprina larval preparations that are unlikely to be exposed to the sheep's immune system during a normal infection. Antibodies were successfully produced against a crude third instar midgut homogenate preparation (MG), and Sodium dodecyl sulphate (SDS) and Triton X-114 (TX) detergent extracts of first instar larvae. Characterisation of the relevant antigens was performed using 1- and 2D gel electrophoresis and immunoblotting, immunoperoxidase histological studies and in vitro larval growth cultures. All the mAbs were of the IgM isotype. Common recognition of bands at 40, 50 and 80 kDA was evident on 1D blots of larval organ preparations by most mAbs while recognition of antigens in the 2D blots appeared to be more specific. Immunohistological studies suggested that a number of the antibodies specifically bound to intracellular structures within the midgut epithelium. However, antibodies derived from one clone also recognised the epithelium of Malpighian tubules, oenocytes and muscle fibres. None of the antibodies raised against TX extracts were observed to bind to larval structures. Results of larval cultures suggested that certain antibodies could significantly inhibit larval survival and growth in vitro.
Fry
K L
KL
School of Agriculture, La Trobe University, Bundoora, Victoria, Australia.
Seaton
D S
DS
Sandeman
R M
RM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000941:Antigens / Q000032:analysis / Q000276:immunology; D004175:Diptera / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D007150:Immunohistochemistry; D007814:Larva / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D000941:Antigens