Weitzman S A, Desmond M C, Stossel T P
J Clin Invest. 1979 Jul;64(1):321-5. doi: 10.1172/JCI109455.
Turnover of membrane constituents appears important in many biologic processes. We studied this process in neutrophils by immunologic methods. The capacity of neutrophils to recognize other neutrophils coated with antibodies against membrane antigens was used to determine the changes that occur after attachment of the antibody to the neutrophil membrane. Neutrophils were sensitized for 30 min at 22 degrees C with antibodies from three patients with antineutrophil autoantibodies. The sensitized neutrophils were recognized by normal neutrophils, which responded with an increase in glucose oxidation. If, after sensitization, the sensitized neutrophils were not immediately exposed to normal neutrophils, but instead were incubated at 37 degrees C for varying times, the capacity to elicit a recognition response decayed and was gone by 30 min. Additionally, the capacity of the cells to be resensitized by reexposure to antibody also decayed during this period. However, after further incubation at 37 degrees C, the neutrophils recovered the capacity to become sensitized; and this recovery was not inhibited by the addition of cycloheximide. Control incubations with normal immunoglobulin (Ig)G did not elicit a recognition response. The decay in recognition response was temperature dependent. Direct immunofluorescent studies with fluorescein-conjugated antineutrophil IgG revealed that the antibodies were cleared by aggregation and endocytosis. We conclude that: (a) neutrophils clear antibody from the cell surface by a temperature-dependent mechanism; (b) antigenicity is cleared concomitantly; (c) the mechanism of clearance involves internalization; and (d) with time, antigenicity reappears on the cell surface.
膜成分的更新在许多生物过程中似乎都很重要。我们通过免疫学方法研究了中性粒细胞中的这一过程。利用中性粒细胞识别被抗膜抗原抗体包被的其他中性粒细胞的能力,来确定抗体附着于中性粒细胞膜后发生的变化。用来自三名抗中性粒细胞自身抗体患者的抗体,在22℃下将中性粒细胞致敏30分钟。致敏的中性粒细胞可被正常中性粒细胞识别,后者会通过增加葡萄糖氧化做出反应。如果在致敏后,致敏的中性粒细胞没有立即与正常中性粒细胞接触,而是在37℃下孵育不同时间,引发识别反应的能力会衰减,30分钟后就会消失。此外,在此期间,细胞通过再次接触抗体而重新致敏的能力也会衰减。然而,在37℃下进一步孵育后,中性粒细胞恢复了致敏能力;并且这种恢复不受添加放线菌酮的抑制。用正常免疫球蛋白(Ig)G进行的对照孵育未引发识别反应。识别反应的衰减与温度有关。用荧光素偶联的抗中性粒细胞IgG进行的直接免疫荧光研究表明,抗体通过聚集和内吞作用被清除。我们得出以下结论:(a)中性粒细胞通过温度依赖性机制从细胞表面清除抗体;(b)抗原性同时被清除;(c)清除机制涉及内化作用;(d)随着时间的推移,抗原性会重新出现在细胞表面。