Eschenbach C, Hahm W, Adrian U
Klin Wochenschr. 1985 Apr 15;63(8):364-70. doi: 10.1007/BF01731656.
We have described a method to measure the chemiluminescence of stimulated monocytes, which necessitated only a few ml blood and thus make possible the investigation of clinical problems. The measurement of the chemiluminescence of monocytes is performed in a suspension of mononuclear leukocytes containing 40%-50% monocytes and 50%-60% lymphocytes. The preparation of this suspension requires a smaller blood volume than that of a suspension of pure monocytes. This method is possible because of the incapability of lymphocytes to produce chemiluminescence. A further important element of the described procedure is the fact that contamination of the mononuclear cell suspension with polymorphonuclear leukocytes, which show pronounced activity of oxidative metabolism and thus a high degree of chemiluminescence, is excluded. The application of 7 Dimethylamino-naphthalene-1,2-dicarbonic acid hydrazide (DMNH) as highly efficient chemiluminescent oxidant indicator and of the sensitive chemiluminescence measuring instrument Biolumat LB 9 500 permit the assessment of the chemiluminescence activity of monocytes in 10(5) cells. By varying the single components of the measuring approach (number of monocytes, number of ingesting particles, concentration of DMNH) the testing conditions were examined and a standard approach was determined. The average value of the chemiluminescence of phagocytosing monocytes in healthy adults during 30 min measuring time is 3.13 (S = 0.79) X 10(6) relative light units/10(5) cells. The point of maximum photone emission per unit in time is reached after 5-10 min.
我们描述了一种测量受刺激单核细胞化学发光的方法,该方法仅需几毫升血液,从而使临床问题的研究成为可能。单核细胞化学发光的测量是在含有40%-50%单核细胞和50%-60%淋巴细胞的单核白细胞悬液中进行的。制备这种悬液所需的血量比纯单核细胞悬液要少。由于淋巴细胞不能产生化学发光,所以这种方法是可行的。所述程序的另一个重要因素是排除了多形核白细胞对单核细胞悬液的污染,多形核白细胞表现出明显的氧化代谢活性,因此化学发光程度很高。使用7-二甲基氨基萘-1,2-二羧酸酰肼(DMNH)作为高效化学发光氧化剂指示剂以及灵敏的化学发光测量仪器Biolumat LB 9500,可以评估10⁵个细胞中单核细胞的化学发光活性。通过改变测量方法的各个组成部分(单核细胞数量、吞噬颗粒数量、DMNH浓度)来检查测试条件并确定标准方法。在30分钟测量时间内,健康成年人吞噬单核细胞的化学发光平均值为3.13(S = 0.79)×10⁶相对光单位/10⁵个细胞。单位时间内最大光发射点在5-10分钟后达到。