Weiss J, Victor M, Stendhal O, Elsbach P
J Clin Invest. 1982 Apr;69(4):959-70. doi: 10.1172/jci110535.
Previous studies have suggested that a cationic bactericidal/permeability-increasing protein (BPI) present in both rabbit and human polymorphonuclear leukocytes is the principal O2-independent bactericidal agent of these cells toward several strains of Escherichia coli and Salmonella typhimurium (1978. J. Biol. Chem. 253: 2664--2672; 1979. J. Biol. Chem. 254: 11000--11009). To further evaluate the possible role of this protein in the killing of gram-negative bacteria by polymorphonuclear leukocytes, we have measured the bactericidal activity of intact rabbit peritoneal exudate leukocytes under aerobic or anaerobic conditions and of intact human leukocytes from a patient with chronic granulomatous disease. Anaerobic conditions were created by flushing the cells under a nitrogen stream. Effective removal of oxygen was demonstrated by the inability of nitrogen-flushed leukocytes to mount a respiratory burst (measured as increased conversion of 1-[14C]glucose leads to 14CO2 or by superoxide production) during bacterial ingestion. At a bacteria/leukocyte ratio of 10:1, killing of gram-positive, BPI-resistant, Staphylococcus epidermidis is markedly impaired in the absence of oxygen (76.4 +/- 3.3% killing in room air, 29.2 +/- 8.2% killing in nitrogen). Essentially all increased bacterial survival is intracellular. In contrast, both a nonopsonized rough strain (MR-10) and an opsonized smooth strain (MS) of S. typhimurium 395 are killed equally well in room air and nitrogen. A maximum of 70--80 MR-10 and 30--40 MS are killed per leukocyte either in the presence or absence of oxygen. There is no intracellular bacterial survival in either condition indicating that intracellular O2-independent bactericidal system(s) of rabbit polymorphonuclear leukocytes can at least match the leukocyte's ingestive capacity. Whole homogenates and crude acid extracts manifest similar bactericidal capacity toward S. typhimurium 395. This activity can be accounted for by the BPI content of these cell fractions and is virtually eliminated by immune (anti-BPI), but not by preimmune goat IgG-rich fractions. Opsonization of smooth MS, required for bacterial killing by intact leukocytes, does not alter bacterial sensitivity to BPI in crude or purified form. Leukocytes of a patient with chronic granulomatous disease killed ingested S. typhimurium 396 MS nearly as well as did normal leukocytes. The bactericidal activity toward E. coli (J5) of crude acid extracts of the CGD and normal human leukocytes was virtually the same and was nearly completely inhibited by anti-BPI IgG-rich fractions, but not by preimmune IgG-rich fractions. These findings suggest that the killing of gram-negative bacteria such as S. typhimurium by intact polymorphonuclear leukocytes may also be attributed to the action of BPI.
先前的研究表明，兔和人多形核白细胞中存在的一种阳离子杀菌/通透性增加蛋白（BPI）是这些细胞对几种大肠杆菌和鼠伤寒沙门氏菌菌株的主要非氧依赖性杀菌因子（1978年。《生物化学杂志》253:2664 - 2672；1979年。《生物化学杂志》254:11000 - 11009）。为了进一步评估这种蛋白在多形核白细胞杀灭革兰氏阴性菌中的可能作用，我们测量了完整兔腹膜渗出白细胞在有氧或无氧条件下以及一名慢性肉芽肿病患者的完整人白细胞的杀菌活性。通过在氮气流下冲洗细胞来创造无氧条件。在细菌摄取过程中，用氮气冲洗的白细胞无法产生呼吸爆发（以1 - [14C]葡萄糖向14CO2的转化增加或超氧化物产生来衡量），这证明了氧气的有效去除。在细菌/白细胞比例为10:1时，在无氧情况下，革兰氏阳性、对BPI耐药的表皮葡萄球菌的杀灭明显受损（在空气中杀灭率为76.4±3.3%，在氮气中杀灭率为29.2±8.2%）。基本上所有增加的细菌存活都是细胞内的。相比之下，鼠伤寒沙门氏菌395的非调理粗糙菌株（MR - 10）和调理光滑菌株（MS）在空气和氮气中杀灭效果相同。在有氧或无氧情况下，每个白细胞最多可杀灭70 - 80个MR - 10和30 - 40个MS。在这两种情况下都没有细胞内细菌存活，这表明兔多形核白细胞的细胞内非氧依赖性杀菌系统至少可以与白细胞的摄取能力相匹配。全匀浆和粗酸提取物对鼠伤寒沙门氏菌395表现出相似的杀菌能力。这种活性可以由这些细胞组分中的BPI含量来解释，并且实际上被免疫（抗BPI）但不被富含免疫前山羊IgG的组分消除。完整白细胞杀灭细菌所需的光滑MS的调理作用，不会改变细菌对粗制或纯化形式的BPI的敏感性。一名慢性肉芽肿病患者的白细胞杀灭摄取的鼠伤寒沙门氏菌396 MS的效果几乎与正常白细胞一样好。慢性肉芽肿病患者和正常人白细胞的粗酸提取物对大肠杆菌（J5）的杀菌活性几乎相同，并且几乎完全被富含抗BPI IgG的组分抑制，但不被富含免疫前IgG的组分抑制。这些发现表明，完整的多形核白细胞对革兰氏阴性菌如鼠伤寒沙门氏菌的杀灭也可能归因于BPI的作用。
