Characterization of a major surface-associated excretory-secretory antigen of Trichinella spiralis larvae with antibodies to keyhole limpet haemocyanin.
A multi-subunit antigen (native M(r) > 200 kDa, reduced M(r) 97-100 kDa) has been identified in homogenates of Trichinella spiralis larvae using affinity-purified rabbit anti-keyhole limpet haemocyanin (KLH) antibodies and its cross-reactivity with KLH was confirmed by competition blotting. The antigen was not present at the larval surface but was exposed after treatment of the larvae with the detergent cetyltrimethyl ammonium bromide (CTAB) which removed the surface coat. This correlated with a significant decrease in insertion of the surface-restricted fluorescent lipid probe AF18, indicating that the surface coat must be lipidic in nature. Unlike KLH, the larval antigen blotted onto nitrocellulose was itself periodate insensitive. Periodate treatment of whole larvae, however, resulted in shedding of the surface, to which anti-KLH antibodies then bound intensely. Anti-KLH antibodies also recognized three (49, 55, 108 kDa) of the four most dominant antigens in excretory-secretory (ES) products of cultured larvae, whose excretion-secretion was increased with CTAB. The nature, location and function of the antigen is discussed.
Modha
J
J
Department of Biochemistry, University of Glasgow.
Robertson
W M
WM
Kennedy
M W
MW
Kusel
J R
JR
D000818:Animals; D000909:Antibodies, Helminth; D000947:Antigens, Helminth / Q000737:chemistry / Q000302:isolation & purification*; D000954:Antigens, Surface / Q000737:chemistry / Q000302:isolation & purification*; D001667:Binding, Competitive; D000077286:Cetrimonium; D002593:Cetrimonium Compounds; D003429:Cross Reactions; D003902:Detergents; D005452:Fluoresceins; D005455:Fluorescent Antibody Technique; D006433:Hemocyanins / Q000276:immunology; D015151:Immunoblotting; D007814:Larva / Q000276:immunology; D008563:Membrane Lipids / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D051379:Mice; D008970:Molecular Weight; D010504:Periodic Acid; D017160:Trichinella spiralis / Q000276:immunology*
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D000954:Antigens, Surface; D002593:Cetrimonium Compounds; D003902:Detergents; D005452:Fluoresceins; D008563:Membrane Lipids; D010504:Periodic Acid; C034806:5-(octadecylthiocarbamoylamino)fluorescein; D006433:Hemocyanins; C009288:metaperiodate; C032808:keyhole-limpet hemocyanin; D000077286:Cetrimonium