Crossreactivity of hemolytic and hemagglutinating proteins in the coelomic fluid of lumbricidae (Annelida).
Oligospecific antisera against the hemolytic and hemagglutinating compounds of the coelomic fluid (CF) of the earthworm species Eisenia fetida, Lumbricus terrestris and Aporrectodea caliginosa were prepared. The proteins were bound to the membranes of erythrocytes and injected into rabbits. By the use of these antisera the following results were obtained: 1) The antisera RaL T and RaAC reached a titer of 1:64,000, the detection limit of RaEF was 1:512,000. RaEF was demonstrated to be oligospecific only against three hemolytic proteins by Western blotting. 2) RaEF is able to inhibit the biological activity not only of hemolysins (E. fetida, A. caliginosa) but also of hemagglutinins (L. terrestris, L. rubellus, D. rubidus). 3) Complex carbohydrates, but not simple sugar compounds, are able to inhibit hemolytic and agglutinating activities in the CF of the investigated species. Fet. and alpha 1ac. GP were demonstrated to be strong inhibitors both of the hemolytic and of the hemagglutinating activity, whereas N-acetylglucosamine was only able to inhibit the hemagglutinating activity. 4) The investigated compounds were shown to crossreact in ELISA experiments. 5) By the use of Western blotting, the crossreacting molecules in the CF of the investigated lumbricid species were identified.
Mohrig
W
W
University of Texas, M.D. Anderson Cancer Center, Department of Cell Biology, Houston 77030, USA.
Eue
I
I
Kauschke
E
E
Hennicke
F
F
D000818:Animals; D000918:Antibody Specificity; D015153:Blotting, Western; D001826:Body Fluids / Q000737:chemistry / Q000378:metabolism; D002241:Carbohydrates / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006388:Hemagglutinins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006460:Hemolysin Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006462:Hemolytic Plaque Technique; D005779:Immunodiffusion; D009835:Oligochaeta / Q000502:physiology*; D011817:Rabbits / Q000276:immunology
D002241:Carbohydrates; D006388:Hemagglutinins; D006460:Hemolysin Proteins