Van Nerom A, Ducatelle R, Haesebrouck F
Department of Pathology, Bacteriology and Avian Diseases, Faculty of Veterinary Medicine, University of Ghent, Salisburylaan, Merelbeke, Belgium.
Vet Microbiol. 1999 Mar 12;65(3):173-84. doi: 10.1016/s0378-1135(98)00294-6.
Chemiluminescence (CL) was used to investigate the competence of turkey monocytes to mount a respiratory burst response upon interaction with Chlamydia psittaci. The oxidative activity of purified turkey monocytes, following inoculation with the avian C. psittaci serovar D strain 92/1293, was studied using luminol- and lucigenin-enhanced CL. Purified turkey monocytes were inoculated with C. psittaci at multiplicity of infection (MOI) of approximately 100, 10 and 1. In the presence of luminol, no detectable CL or only a weak CL response was obtained, and if present it increased with increasing MOI. Either sham inoculated monocytes, or monocyte-free control assays supplemented with C. psittaci, gave no detectable luminol-enhanced CL responses. In the lucigenin-enhanced assays, monocytes inoculated with C. psittaci demonstrated an immediate CL peak, the height of which was proportional to the MOI used. Following inoculations at a MOI 1, a faint second peak was observed, when applying high concentrations of lucigenin. Sham inoculated monocytes gave no detectable lucigenin-enhanced CL responses. However, in the presence of lucigenin, the addition of C psittaci to monocyte-free controls also resulted in an immediate CL peak, though no second peak was detected. This immediate lucigenin-dependent CL peak induced by C. psittaci was similar to the one observed in the presence of monocytes, and was not inhibited by superoxide dismutase. We demonstrated that this avian C. psittaci strain induces only a very weak respiratory burst response in turkey monocytes. In contrast, C. psittaci itself elicited an intense non-superoxide mediated lucigenin-dependent CL, indicating that in chlamydial research the detection of superoxide, using lucigenin, should be confirmed with a specific superoxide inhibitor.
采用化学发光法（CL）研究火鸡单核细胞与鹦鹉热衣原体相互作用时引发呼吸爆发反应的能力。使用鲁米诺和光泽精增强的化学发光法，研究接种禽源鹦鹉热衣原体血清型D菌株92/1293后纯化的火鸡单核细胞的氧化活性。以约100、10和1的感染复数（MOI）用鹦鹉热衣原体接种纯化的火鸡单核细胞。在鲁米诺存在的情况下，未检测到化学发光或仅获得微弱的化学发光反应，若有反应则随MOI增加而增强。无论是假接种的单核细胞，还是补充了鹦鹉热衣原体的无单核细胞对照试验，均未检测到鲁米诺增强的化学发光反应。在光泽精增强试验中，接种鹦鹉热衣原体的单核细胞呈现立即出现的化学发光峰值，其高度与所用的MOI成正比。以MOI 1接种后，当使用高浓度光泽精时，观察到一个微弱的第二个峰值。假接种的单核细胞未检测到光泽精增强的化学发光反应。然而，在光泽精存在的情况下，向无单核细胞对照中添加鹦鹉热衣原体也导致立即出现化学发光峰值，尽管未检测到第二个峰值。鹦鹉热衣原体诱导的这种立即出现的依赖光泽精的化学发光峰值与在单核细胞存在下观察到的相似，且不受超氧化物歧化酶抑制。我们证明，这种禽源鹦鹉热衣原体菌株在火鸡单核细胞中仅诱导非常微弱的呼吸爆发反应。相比之下，鹦鹉热衣原体本身引发强烈的非超氧化物介导的依赖光泽精的化学发光，这表明在衣原体研究中，使用光泽精检测超氧化物时，应使用特异性超氧化物抑制剂进行确认。
