Activation of the alternative complement pathway by natural antibody to glycolipids in guinea-pig serum

Abstract

Liposomes containing paragloboside (PG) on their membrane were readily lysed by C4-deficient guinea-pig serum (C4D-GPS) through activation of the alternative complement pathway (ACP). Therefore we examined the reactivity of several types of guinea-pig serum (GPS) on PG-liposomes and determined that all GPS except that from specific pathogen-free (SPF) Hartley guinea-pigs had lytic capacity in Mg-EGTA-GVB (gelatin veronal-buffered saline containing Mg++ and ethyleneglycol-bis(beta-aminoethyl ether)N,N'-tetraacetate). This lytic capacity of GPS corresponded with the amount of natural antibody to PG in those sera. Although GPS of SPF guinea-pigs (SPF-GPS) could not lyse PG-liposomes in Mg-EGTA-GVB, it could lyse the liposomes when heated C4D-GPS or Hartley GPS was added. Natural antibody to PG in the heated sera was regarded to have sensitized PG-liposomes to lysis by SPF-GPS via ACP activation. Since the antibody to PG-liposomes was removed by lacto-N-nor-hexaosylceramide which has the same chemical structure in the terminal oligosaccharide, the antibody to PG in GPS was suggested to have a specificity to the terminal structure of oligosaccharide shared by lacto-N-nor-hexaosylceramide. Furthermore, the IgM fraction, which had been prepared by gel filtration of heated C4D-GPS on a Sephadex G200 column, could also sensitize PG-liposomes to lytic reaction of SPF-GPS in Mg-EGTA-GVB. This sensitizing capacity of heated C4D-GPS was suppressed by absorption of the serum or its IgM fraction with anti-guinea-pig mu-chain antibody coupled to Sepharose. Therefore, it was concluded that the lysis of PG-liposomes by GPS in Mg-EGTA-GVB was a result of ACP activation mediated by natural antibodies to PG of the IgM type which are present in usual GPS. This conclusion indicated that natural antibodies of the IgM type might play a role with ACP in host defence, especially in C4-deficient guinea-pigs where the classical complement pathway is impaired.