PRODUCTION OF ANTIBODY AGAINST HUMAN LEUKOCYTIC PYROGEN
PRODUCTION OF ANTIBODY AGAINST HUMAN LEUKOCYTIC PYROGEN
复制标题
DOI：
10.1172/jci108797
复制
发表时间：
1977-01-01
影响因子：
15.9
通讯作者：
WOLFF, SM
中科院分区：
文献类型：
--
作者：
DINARELLO, CA;RENFER, L;WOLFF, SM
体外用杀死的葡萄球菌刺激人外周血白细胞释放白细胞热原（LP）。将这些受刺激的白细胞的上清浓缩，在弗氏完全佐剂中乳化，然后皮下注射到家兔体内。经过7个月的强化注射，兔抗血清破坏了人LP的热原活性，并且在随后的7个月里，这种中和能力的滴度增加。兔抗血清的热原中和能力在Sephadex G-200的球蛋白部分、免疫球蛋白G和IgM峰以及山羊抗兔IgG免疫吸附剂的酸洗脱部分中恢复。中和抗体对兔、豚鼠、猴LP无影响，对人LP有特异性。当与Sepharose偶联时，该抗体结合人LP；从免疫吸附剂中酸洗脱后，LP被回收，而没有失去生物或化学特性。抗血清被不含LP的刺激白细胞上清吸收，这对抗LP的效价没有影响。从吸收抗血清制备的免疫吸附剂柱中洗脱的粗LP，经聚丙烯酰胺凝胶电泳评估，其含有显著减少的污染蛋白。这些研究证实可以产生针对人白细胞热原的特异性抗体。该抗体对进一步研究和纯化白细胞热原及其在人发热发病机制中的作用具有重要意义。
Human peripheral blood leukocytes were stimulated with killed staphylococci in vitro to release leukocytic pyrogen (LP). Supernates from these stimulated leukocytes were concentrated, emulsified in Freund''s complete adjuvant and injected intradermally into rabbits. After 7 monthly booster injections, rabbit antiserum destroyed the pyrogenic activity of human LP, and the titer of this neutralizing ability increased in the subsequent 7 mo. The pyrogen-neutralizing capacity of the rabbit antiserum was recovered in the globulin fraction, the Ig[immunoglobulin]G and IgM peaks of Sephadex G-200, and the acid-eluted fraction of a goat anti-rabbit IgG immunoadsorbant. The neutralizing antibody was specific for human LP inasmuch as it had no effect on rabbit, guinea pig or monkey LP. When coupled to Sepharose, this antibody bound human LP; after acid elution from this immunoadsorbant, LP was recovered without loss of biologic or chemical characteristics. The antiserum was absorbed with stimulated leukocyte supernates which did not contain LP, and this had no effect on the titer of anti-LP. Crude human LP, eluted from immunoadsorbant columns prepared from absorbed antiserum, contained significantly reduced contaminating protein when evaluated by polyacrylamide gel electrophoresis. These studies established that specific antibody to human leukocytic pyrogen can be produced. This antibody is useful in the further study and pruification of luekocytic pyrogen and its role in the pathogenesis of human fever.