Immunogenic properties of mannose-containing ceramide disaccharide and immunochemical detection of its hapten in the two kinds of crustacean, Euphausia superba and Macrobrachium nipponense.
Antiserum against Man beta 1-4Glc beta 1-1Ceramide (MIOse2Cer), a mannolipid isolated from spermatozoa of the fresh-water bivalve, Hyriopsis schlegellii, has been elicited in rabbits by repeated injection of a mixture of hapten-bovine serum albumin (1:1, mg/ml) with Freund's adjuvant. The specificity of the affinity-purified antibody (immunoglobulin G type) obtained from the serum was examined, using other glycosphingolipids and glyco-proteins structurally related to MIOse2Cer, by means of ELISA and TLC-immunostaining. The purified antibody was highly specific to MIOse2Cer and lacked reactivity with other glycolipids and glycoproteins including glucosylceramide, lactosylceramide, dimannosylglucosylceramide (MIOse3Cer), glucosaminylmannosylglucosylceramide (ArOse3Cer), thyroglobulin and alpha 1-acid glycoprotein. The antibody was found to bind, although less efficiently, to certain other compounds containing the group Man beta 1-4Glc and/or Man beta 1-4GlcNAc at their termini, such as MIOse2-sphingosine and Man beta 1-4GlcNAc beta 1-p-aminobenzoic acid ethylester derivatives. The present antibody was applied to the detection of the natural hapten in crustacean glycolipids. The purified antibody reacted with a neutral glycosphingolipid present in the two kinds of crustacean, Euphausia superba (antarctic krill) and Macrobrachium nipponense (fresh-water shrimp) as shown by TLC-immunostaining. The crustacean glycolipid antigen was isolated and characterized to be the Man beta 1-4Glc-Cer. This is the first report on the presence of a mannose-containing glycosphingolipid in the crustacean.
Itonori
S
S
Department of Chemistry, Faculty of Liberal Arts and Education, Shiga University, Japan.
Hiratsuka
M
M
Sonku
N
N
Tsuji
H
H
Itasaka
O
O
Hori
T
T
Sugita
M
M
D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000302:isolation & purification; D000918:Antibody Specificity; D002240:Carbohydrate Sequence; D002518:Ceramides / Q000737:chemistry / Q000276:immunology*; D002855:Chromatography, Thin Layer; D003445:Crustacea / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006241:Haptens / Q000276:immunology*; D015151:Immunoblotting; D008297:Male; D008358:Mannose / Q000737:chemistry; D008969:Molecular Sequence Data; D033541:Palaemonidae / Q000276:immunology
D000906:Antibodies; D002518:Ceramides; D006241:Haptens; D008358:Mannose