Omata M, Watanabe M, Hirasawa N, Tsurufuji S, Mue S, Ohuchi K
Department of Biochemistry, Faculty of Pharmaceutical Sciences, Tohoku University, Miyagi, Japan.
J Clin Lab Immunol. 1990 Apr;31(4):175-81.
The number of neutrophils, eosinophils, and mononuclear cells that had migrated into pouch fluid were measured after injecting an antigen (azobenzenearsonate-conjugated acetyl bovine serum albumin) solution into air pouches made on the dorsum of immunized rats. The number of neutrophils began to increase 4 hr after the antigen challenge, reaching a maximum at 16-24 hr. Increases in the number of eosinophils and mononuclear cells were quite poor. The pouch fluid supernatant fraction was fractionated to hydrophilic and lipophilic fractions with the aid of an octadecylsilyl silica cartridge, and the chemotactic activity in each fraction was measured by the modified Boyden chamber method. The chemotactic activity in the pouch fluid supernatant fraction was the highest at 2-4 hr and decreased with time after the antigen challenge. At 16 and 24 hr, chemotactic activity was decreased to a very low level although there was a large number of neutrophils in the pouch fluid. At 4 hr, the chemotactic activity in the lipophilic fraction was significantly higher than that in the hydrophilic fraction. However, at 8 hr, the chemotactic activity in the lipophilic fraction was changed to a significantly lower level than that in the hydrophilic fraction. The chemotactic activity in the lipophilic fraction had decreased to a very low level at 8 hr. Heat treatment of the pouch fluid supernatant fraction at 92 degrees C for 5 min both at 4 and 8 hr significantly increased the chemotactic activity.(ABSTRACT TRUNCATED AT 250 WORDS)
将抗原(偶氮苯砷酸盐结合的乙酰牛血清白蛋白)溶液注入免疫大鼠背部的气袋后,测定迁移到袋液中的中性粒细胞、嗜酸性粒细胞和单核细胞的数量。抗原攻击后4小时,中性粒细胞数量开始增加,在16 - 24小时达到峰值。嗜酸性粒细胞和单核细胞数量的增加很少。借助十八烷基硅烷硅胶柱将袋液上清部分分离成亲水和亲脂部分,并用改良的博伊登室法测定各部分的趋化活性。抗原攻击后,袋液上清部分的趋化活性在2 - 4小时最高,并随时间下降。在16和24小时,尽管袋液中有大量中性粒细胞,但趋化活性降至非常低的水平。在4小时时,亲脂部分的趋化活性明显高于亲水部分。然而,在8小时时,亲脂部分的趋化活性变为明显低于亲水部分。在8小时时,亲脂部分的趋化活性已降至非常低的水平。在4小时和8小时时,将袋液上清部分在92℃加热5分钟均显著提高了趋化活性。(摘要截短至250字)