Phagocytosis and degradation of DNA‐anti‐DNA complexes by human phagocytes I. Assay conditions, quantitative aspects and differences between human blood monocytes and neutrophils

Phagocytosis and degradation of DNA‐anti‐DNA complexes by human phagocytes I. Assay conditions, quantitative aspects and differences between human blood monocytes and neutrophils
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人吞噬细胞对 DNA-抗 DNA 复合物的吞噬和降解 I. 人血单核细胞和中性粒细胞的测定条件、定量方面以及差异

DOI:
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发表时间:
1981
影响因子:
5.4
通讯作者:
D. Roos
D. Roos
中科院分区:
医学3区
文献类型:
--
作者:
M. Lamers;Elisabeth R. de Groot;D. Roos

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研究了人血液中的中性粒细胞和单核细胞对 3H 标记的 DNA-抗 DNA 复合物的体外摄取。复合物由噬菌体 PM2 的 3H 标记的环状双链 (ds) DNA 和系统性红斑狼疮患者的含抗 dsDNA 血清制备。吞噬作用后,分离细胞和培养基。在对细胞相关放射性进行计数之前,用 DNase 处理细胞以去除粘附和未摄入的复合物。因此,仅测量细胞内的复合物。分析介质的酸沉淀放射性。通过这种方式,我们发现中性粒细胞仅吞噬复合物,而单核细胞吞噬复合物并降解抗原。相反,两种类型的吞噬细胞都会降解破伤风-抗破伤风复合物中的抗原。降解发生在细胞内的吞噬作用之后。中性粒细胞和单核细胞之间 DNA 降解的差异与溶酶体部分的酸性 DNase 活性的差异相关:单核细胞含有 DNase 活性,而中性粒细胞则不含有。利用 DNA 与 131I 标记的抗 DNA 制成的复合物,我们发现两种细胞类型都会降解抗体。复合物的摄取和抗原的降解随着孵育时间和细胞浓度的增加而增加,并且相对于复合物浓度而言是饱和的。该过程受到 5 mM 单碘乙酸或低温的抑制。
The uptake in vitro was studied of 3H‐labeled DNA‐anti‐DNA complexes by neutrophils and monocytes from human blood. Complexes were prepared from 3H‐labeled circular double‐stranded (ds) DNA of bacteriophage PM2 and anti‐dsDNA‐containing sera from patients with systemic lupus erythematosus. After phagocytosis, cells and medium were separated. The cells were treated with DNase to remove adherent and noningested complexes before the cell‐associated radioactivity was counted. Thus, only complexes inside the cells were measured. The medium was analyzed for acid‐precipitable radioactivity. In this way, we found that neutrophils only phagocytose the complexes, whereas monocytes phagocytose the complexes and degrade the antigen. In contrast, both types of phagocyte degraded the antigen in tetanus‐antitetanus complexes. The degradation took place after phagocytosis, inside the cells. The difference in DNA degradation between neutrophils and monocytes correlated with the difference in acid DNase activity of the lysosomal fractions: monocytes contained DNase activity, neutrophils did not. With complexes made from DNA with 131I‐labeled anti‐DNA, we found that both cell types degraded the antibody. Uptake of complexes and degradation of antigen increased with incubation time and cell concentration and was saturable with respect to complex concentration. The processes were inhibited by 5 mM mono‐iodoacetic acid or by low temperatures.