Lopes Alves L, Travassos L R, Previato J O, Mendonça-Previato L
Departamento de Biologia Celular e Genética, UERJ, Brazil.
Glycobiology. 1994 Jun;4(3):281-8. doi: 10.1093/glycob/4.3.281.
Antisera were raised in rabbits against acetone-dried yeast-like and mycelium forms of Sporothrix schenckii. These antisera were tested for immunoprecipitation of peptidorhamnomannans isolated from both cell types. Both antisera reacted strongly with S.schenckii peptidorhamnomannans, but the reactions were weak with beta-eliminated peptidopolysaccharides. These antisera did not recognize the Saccharomyces cerevisiae mannoprotein, and reacted poorly with Ceratocystis (Ophiostoma) stenoceras cell wall glycopeptides. Since beta-eliminated glycopeptides were poorly reactive, we investigated the activity of O-glycosidically linked oligosaccharides which were liberated from the peptidorhamnomannans by mild alkaline hydrolysis, using a hapten inhibition test. The rates of inhibition showed that the immunodominant epitopes in O-linked tetra- and pentasaccharides had the following novel structures: alpha-L-Rhap (1-->4)-alpha-D-GlcpA and alpha-L-Rhap(1-->4)-[alpha-L-Rhap(1-->2)]alpha-D-GlcpA . Results suggest that peptidorhamnomannans or synthetic antigens bearing these epitopes should be used for the specific detection of anti-S.schenckii antibodies rather than the long-chain N-linked polysaccharides.
用丙酮干燥的申克孢子丝菌酵母样和菌丝体形式免疫家兔制备抗血清。检测这些抗血清对从两种细胞类型中分离的肽甘露聚糖的免疫沉淀作用。两种抗血清均与申克孢子丝菌肽甘露聚糖强烈反应,但与β-消除的肽聚糖反应较弱。这些抗血清不能识别酿酒酵母甘露糖蛋白,与短喙壳属(长喙壳属)细胞壁糖肽反应较差。由于β-消除的糖肽反应性较差,我们使用半抗原抑制试验研究了通过温和碱性水解从肽甘露聚糖中释放的O-糖苷键连接的寡糖的活性。抑制率表明,O-连接的四糖和五糖中的免疫显性表位具有以下新结构:α-L-鼠李糖(1→4)-α-D-葡萄糖醛酸和α-L-鼠李糖(1→4)-[α-L-鼠李糖(1→2)]α-D-葡萄糖醛酸。结果表明,带有这些表位的肽甘露聚糖或合成抗原应用于抗申克孢子丝菌抗体的特异性检测,而不是长链N-连接的多糖。