Vinh T, Faine S, Handley C J, Adler B
Department of Microbiology, Monash University, Clayton, Victoria, Australia.
FEMS Immunol Med Microbiol. 1994 Feb;8(2):99-107. doi: 10.1111/j.1574-695X.1994.tb00431.x.
Leptospiral lipopolysaccharides (LPS) are the main antigens responsible for immunity in leptospirosis. In this investigation we studied the nature of the antigenic determinants of LPS extracted from Leptospira interrogans serovar hardjo (reference strain Hardjoprajitno). The reactions of anti-LPS monoclonal antibodies (mAbs) MUM/F1-4/hardjo (IgM) and MUM/F1-6/hardjo (IgG) with whole cell lysates in Western immunoblotting analysis were unaffected by proteinase K treatment. Periodate treatment of the LPS destroyed the binding of MUM/F1-6/hardjo but preserved that of MUM/F1-4/hardjo. Alkaline phosphatase decreased significantly the binding of MUM/F1-4/hardjo to the LPS but only slightly that of MUM/F1-6/hardjo. On the other hand, phosphodiesterase totally destroyed the binding capacity of both monoclonal antibodies in enzyme immunoassays (EIA). A number of mono- and oligosaccharides was used in EIA inhibition studies. Mannose-6-phosphate and galactose-6-phosphate inhibited the binding of MUM/F1-4/hardjo (50% inhibition at a concentration of 5 mM) to the antigen, but glucose-6-phosphate did not. Galactosamine and mannosamine inhibited the binding of MUM/F1-6/hardjo (50% inhibition at a concentration of 3-4 mM), whereas only a weak inhibition was observed with glucosamine. In contrast, N-acetylated amino sugars did not show any inhibition. An O-acetyl group also appears to be involved in the antigen-antibody binding process.
钩端螺旋体脂多糖（LPS）是钩端螺旋体病免疫反应的主要抗原。在本研究中，我们研究了从问号钩端螺旋体血清型哈德乔（参考菌株Hardjoprajitno）中提取的LPS抗原决定簇的性质。在蛋白质印迹分析中，抗LPS单克隆抗体（mAb）MUM/F1-4/hardjo（IgM）和MUM/F1-6/hardjo（IgG）与全细胞裂解物的反应不受蛋白酶K处理的影响。LPS经高碘酸盐处理后，破坏了MUM/F1-6/hardjo的结合，但保留了MUM/F1-4/hardjo的结合。碱性磷酸酶显著降低了MUM/F1-4/hardjo与LPS的结合，但仅轻微降低了MUM/F1-6/hardjo的结合。另一方面，磷酸二酯酶在酶免疫分析（EIA）中完全破坏了两种单克隆抗体的结合能力。在EIA抑制研究中使用了多种单糖和寡糖。6-磷酸甘露糖和6-磷酸半乳糖抑制了MUM/F1-4/hardjo（在5 mM浓度下50%抑制）与抗原的结合，但6-磷酸葡萄糖没有。半乳糖胺和甘露糖胺抑制了MUM/F1-6/hardjo（在3-4 mM浓度下50%抑制）的结合，而葡糖胺仅观察到微弱的抑制。相比之下，N-乙酰化氨基糖没有显示出任何抑制作用。一个O-乙酰基团似乎也参与了抗原-抗体结合过程。
