Beachey E H, Stollerman G H, Johnson R H, Ofek I, Bisno A L
J Exp Med. 1979 Oct 1;150(4):862-77. doi: 10.1084/jem.150.4.862.
We tested the ability of pepsin-extracted, highly purified M protein to induce type-specific immunity in experimental animals and humans. M protein was prepared from limited peptic digests of whole group A type 24 streptococci and was purified to chemical homogeneity as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, quantitative amino acid analysis, and Edman degradation. For vaccination, the lyophilized M24 protein preparation (pep M24) was precipitated in aluminum hydroxide. When injected into laboratory animals, alum-precipitated pep M24 produced type-specific protective antibodies and was free of non-type-specific immunoreactivity. In man, skin tests with 1-microgram doses of pep M24 were negative in all 37 adults tested. 12 adult human volunteers received two-four subcutaneous injections of 100-200 micrograms of alum-precipitated pep M24 at intervals of at least 2 wk. The immune response to pep M24 was measured by a variety of assays designed to detect (a) type-specific humoral antibodies (opsonophagocytic, long chain, and mouse protection tests); (b) total humoral antibodies (complement fixation and enzyme-linked immunosorbent assay); (c) cellular immunity (skin tests); and (d) heart cross-reactive antibodies (immunofluorescence). Type-specific opsonic antibodies developed in 10 of the 12 vaccinees, and positive delayed-type skin tests developed in 11. Immune sera from two of the vaccinees were effective in mouse-protection tests against challenge with M24 but not M6 streptococci. None of the volunteers developed heart-reactive antibodies or antibodies to non-type-specific M protein antigens. Alum-precipitated pep M24 was well-tolerated in man, and no serious local or systemic reactions were observed. Thus, pep M24 induces type-specific, protective antibodies in doses that are well-tolerated in man.
我们测试了经胃蛋白酶提取的高度纯化M蛋白在实验动物和人类中诱导型特异性免疫的能力。M蛋白是从A群24型全链球的有限胃蛋白酶消化物中制备的，并通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、定量氨基酸分析和埃德曼降解判断其纯化至化学纯一性。用于疫苗接种时，冻干的M24蛋白制剂（胃蛋白酶处理的M24，pep M24）沉淀于氢氧化铝中。当注入实验动物时，明矾沉淀的pep M24产生型特异性保护性抗体，且无非型特异性免疫反应性。在人体中，对37名成年人进行1微克剂量的pep M24皮肤试验均为阴性。12名成年志愿者每隔至少2周接受两到四次皮下注射100 - 200微克明矾沉淀的pep M24。通过多种检测方法测量对pep M24的免疫反应，这些检测方法旨在检测：（a）型特异性体液抗体（调理吞噬、长链和小鼠保护试验）；（b）总体液抗体（补体结合和酶联免疫吸附测定）；（c）细胞免疫（皮肤试验）；以及（d）心脏交叉反应抗体（免疫荧光）。12名接种者中有10人产生了型特异性调理抗体，11人出现了阳性迟发型皮肤试验。两名接种者的免疫血清在针对M24而非M6链球菌攻击的小鼠保护试验中有效。志愿者均未产生心脏反应性抗体或针对非型特异性M蛋白抗原的抗体。明矾沉淀的pep M24在人体中耐受性良好，未观察到严重的局部或全身反应。因此，pep M24能以人体耐受性良好的剂量诱导型特异性保护性抗体。
