Andoh A, Fujiyama Y, Hirotani S, Hodohara K, Bamba T, Hosoda S
The Second Department of Internal Medicine, Shiga University of Medical Science, Otsu.
Nihon Ketsueki Gakkai Zasshi. 1990 May;53(3):567-74.
We analyzed the phagocytic activity of purified human monocytes using fluorescent latex beads sensitized with IgG or IgG.C3 by flow cytometry. To prepare IgG-sensitized latex beads (BA), BSA-coated latex beads (B) were incubated with diluted rabbit IgG anti-BSA. To bind complement components, BA were incubated with whole serum pretreated with K-76 monocarboxylic acid (K-76COOH). K-76COOH inhibits the activity of factor I and C5, resulting in deposition of C1, C4b, C2a, C3b on BA (BAC). Phagocytic activity was assessed by percent phagocytosis and phagocytic index (PI). To eliminate the effects of non-phagocytosed latex beads, subtraction of the data at 4 degrees C from 37 degrees C was performed. Percent phagocytosis for 60 min. was B 5.0%, BA 18.3%, and BAC 57.5%, and PI (ingested latex beads/100 cells) was B 7.9, BA 36.8, and BAC 152.7, respectively. In addition, K-76COOH caused dose dependent inhibition on IgG.C3 mediated phagocytosis. Comparison of inhibition pattern on BAC and BA indicated that K-76COOH directly inhibited C3.C3-receptor binding.
我们通过流式细胞术，使用经IgG或IgG.C3致敏的荧光乳胶珠，分析了纯化的人单核细胞的吞噬活性。为制备IgG致敏乳胶珠（BA），将牛血清白蛋白包被的乳胶珠（B）与稀释的兔抗牛血清白蛋白IgG一起孵育。为结合补体成分，将BA与用K-76单羧酸（K-76COOH）预处理的全血清一起孵育。K-76COOH抑制因子I和C5的活性，导致C1、C4b、C2a、C3b沉积在BA上（BAC）。通过吞噬百分比和吞噬指数（PI）评估吞噬活性。为消除未吞噬乳胶珠的影响，进行了37℃数据减去4℃数据的操作。60分钟的吞噬百分比分别为：B为5.0%，BA为18.3%，BAC为57.5%；PI（摄取的乳胶珠/100个细胞）分别为：B为7.9，BA为36.8，BAC为152.7。此外，K-76COOH对IgG.C3介导的吞噬作用产生剂量依赖性抑制。对BAC和BA抑制模式的比较表明，K-76COOH直接抑制C3.C3受体结合。
