Hart P H, Spencer L K, Nulsen M F, McDonald P J, Finlay-Jones J J
Infect Immun. 1986 Mar;51(3):936-41. doi: 10.1128/iai.51.3.936-941.1986.
Intraabdominal abscesses were induced in mice by intraperitoneal inoculation of Bacteroides fragilis and Escherichia coli plus bran as the abscess-potentiating agent. Six- or seven-day-old abscesses were mechanically disaggregated in buffer, and the cells obtained were fractionated on discontinuous Percoll density gradients. Neutrophil populations of different density, each approximately 90% pure, were isolated. When the abscess-derived neutrophils were subsequently incubated with normal serum in vitro under aerobic conditions, the viability of the gram-negative bacteria that had been phagocytosed within the abscess did not change significantly. This anergy to intracellular bacteria (on subsequent incubation in vitro under optimal conditions for phagocytic killing) was also found for neutrophils that had been obtained from abscesses induced by a mixture that included Proteus mirabilis plus B. fragilis and from those induced by E. coli plus P. mirabilis. While unable to significantly kill intracellular organisms that had been phagocytosed in vivo, the abscess-derived neutrophils could engulf and kill organisms to which they were exposed in vitro. Neutrophils from abscesses induced by P. mirabilis only plus bran killed that organism introduced in vitro significantly more effectively than the organisms that had been engulfed in vivo. In contrast, neutrophils from abscesses induced by the gram-positive organism Staphylococcus aureus plus bran were able to kill their intracellular organisms on subsequent incubation in vitro as effectively as they could kill added S. aureus. Neutrophils isolated from the peripheral blood and from induced peritoneal exudates of abscess-bearing mice were able to phagocytose and kill organisms in vitro with greater efficiency than abscess-derived neutrophils. The mechanism whereby neutrophils from abscesses induced by the gram-positive organism S. aureus can kill the organisms phagocytosed in vivo on subsequent in vitro incubation, in contrast to the relative anergy to their intracellular organisms displayed by neutrophils derived from abscesses induced by combinations of gram-negative bacteria, is not known.
通过腹腔接种脆弱拟杆菌、大肠埃希菌并加入麸皮作为脓肿增强剂,在小鼠体内诱导形成腹腔脓肿。将6日龄或7日龄的脓肿在缓冲液中机械分散,所得细胞在不连续的Percoll密度梯度上进行分级分离。分离出不同密度的中性粒细胞群体,每个群体纯度约为90%。随后,将脓肿来源的中性粒细胞在有氧条件下与正常血清进行体外孵育时,脓肿内被吞噬的革兰氏阴性菌的活力没有显著变化。对于从由奇异变形杆菌加脆弱拟杆菌混合物诱导形成的脓肿以及由大肠埃希菌加奇异变形杆菌诱导形成的脓肿中获得的中性粒细胞,在体外最佳吞噬杀伤条件下随后孵育时,也发现了对细胞内细菌的这种无反应性(无能)。虽然脓肿来源的中性粒细胞无法显著杀死体内吞噬的细胞内生物体,但它们能够吞噬并杀死体外接触的生物体。仅由奇异变形杆菌加麸皮诱导形成的脓肿中的中性粒细胞,对体外引入的该生物体的杀伤效果明显优于体内吞噬的生物体。相比之下,由革兰氏阳性菌金黄色葡萄球菌加麸皮诱导形成的脓肿中的中性粒细胞,在随后的体外孵育中能够像杀死添加的金黄色葡萄球菌一样有效地杀死其细胞内生物体。从外周血以及患有脓肿小鼠的诱导性腹腔渗出物中分离出的中性粒细胞,在体外吞噬和杀死生物体的效率高于脓肿来源的中性粒细胞。与革兰氏阴性菌组合诱导形成的脓肿来源的中性粒细胞对其细胞内生物体表现出的相对无反应性相比,革兰氏阳性菌金黄色葡萄球菌诱导形成的脓肿中的中性粒细胞能够在随后的体外孵育中杀死体内吞噬的生物体,其机制尚不清楚。