Adam A, Ciorbaru R, Petit J F, Lederer E
Proc Natl Acad Sci U S A. 1972 Apr;69(4):851-4. doi: 10.1073/pnas.69.4.851.
Trypsin- and chymotrypsin-treated delipidated cell walls of Mycobacterium smegmatis were digested overnight with lysozyme. The water-soluble products thus obtained were filtered on a column of Sephadex G-50; the first peak, excluded from the column, has immunological adjuvant activity. The material of the excluded peak is obtained after lyophilization as a snow-white, fluffy material, soluble in water and insoluble in organic solvents. It behaves as a slightly polydisperse macromolecule in an ultracentrifuge, with an approximate molecular weight of 20,000. All the constituents of this material are typical bacterial cell-wall constituents; thus, the water-soluble adjuvant is considered to be an "oligomer" of the cell wall. When added to Freund's incomplete adjuvant with an antigen (e.g., ovalbumin) and injected into hind-foot pads of guinea pigs, this water-soluble adjuvant increases the amount of precipitating antibodies and induces hypersensitivity to ovalbumin and the biosynthesis of gamma(2)-type precipitating antibodies. The water-soluble material has a stronger adjuvant activity than equal amounts of whole bacteria, cell walls, or wax D, and seems to be the first well-defined, water-soluble, adjuvant-active fraction isolated from Mycobacteria.
用胰蛋白酶和胰凝乳蛋白酶处理耻垢分枝杆菌的脱脂细胞壁，然后用溶菌酶消化过夜。将由此获得的水溶性产物在葡聚糖G - 50柱上过滤；从柱中被排除的第一个峰具有免疫佐剂活性。被排除峰的物质经冻干后得到一种雪白蓬松的物质，可溶于水但不溶于有机溶剂。在超速离心机中，它表现为一种略具多分散性的大分子，分子量约为20,000。该物质的所有成分都是典型的细菌细胞壁成分；因此，这种水溶性佐剂被认为是细胞壁的一种“寡聚物”。当将其与抗原（如卵清蛋白）一起添加到弗氏不完全佐剂中，并注射到豚鼠的后足垫时，这种水溶性佐剂会增加沉淀抗体的量，并诱导对卵清蛋白的超敏反应以及γ(2)型沉淀抗体的生物合成。这种水溶性物质比等量的全菌、细胞壁或蜡D具有更强的佐剂活性，似乎是从分枝杆菌中分离出的第一个明确的、水溶性的、具有佐剂活性的组分。
