Splino M, Mĕrka V, Kyntera F
Zentralbl Bakteriol Orig A. 1976 Aug;235(4):512-20.
The study deals with the phagocytosis of Nocardia asteroides (strain Weipheld) and the subsequent intracellular proliferation in peritoneal macrophage cells. Normal, two-stage-immunized and long-term cortison-treated guinea-pig (28 mg cortison / kg weight / day during 30 days) macrophages were used. Further, the cytotoxic effect of Nocardia upon the cells in the peritoneal washing liquid in vitro and the influence of the normal, immune and antimacrophage serum upon the phagocytosis and the intracellular proliferation were studied. Among the cells obtained from the peritoneal washing liquid macrophages were most frequently subject to phagocytosis, leukocytes to a lesser degree. The normal macrophages phagocytized in 14.56% (Fig. 1), macrophages of two-stage-immunized guinea-pigs in 18.21% (Fig. 2) and macrophages from cortison treated guinea-pigs in 12.48% of cases. Intracellular observation showed phagocytized germs after 3 min. of exposure. The course of the intracellular proliferation of Nocardia can be seen in Fig. 3. The phagocytosis index increases slowly in all three groups of macrophages; least so in the immunized macrophages (1.30-after 8 hours). The highest values were obtained in the macrophages of cortison treated guinea-pigs (2.02-after 8 hours). Within 8 hours of exposure the filaments of Nocardia grew through the cell membrane of phagocytizing cells (Figs. 4 A, 4 B). Fig. 5 shows the course of the cytopathogenic effect of Nocardia upon the cells. After 1 hr. the number of dead cells increased from 0.30% to 1.9-3.8%; after 4 hrs. it reached 8.15-9.80%; after 8 hrs. 10.1-14.80%. The highest values were observed in cells from cortison treated guinea-pigs (14.80%). After addition of normal serum (time of phagocytosis 60 min.) normal peritoneal macrophages phagocytized in 13.30% of cases; immune serum stimulated phagocytosis (16.21%); antimacrophage serum significantly reduced phagocytosis (4.10%). The phagocytosis index in peritoneal macrophages with normal and immune serum increased, reaching values of 1.20-1.58 with in 8 hours; the differences were statistically not significant (Fig. 6). A significant increase of proliferation was observed in peritoneal macrophages with antimacrophage serum (2.07 in 8 hours). Neither normal nor immune guine-pig serum influenced the cytopathogenic effect of Nocardia on the cells of peritoneal washing liquid (Fig. 7). The antimacrophage serum had a rapid and expressive effect upon the macrophages (78.23% of cells died after 2 hours, 100% after 3 hours of exposure).
该研究涉及星状诺卡菌(魏费尔德菌株)的吞噬作用以及随后在腹膜巨噬细胞中的细胞内增殖。使用了正常的、经过两阶段免疫的和长期接受可的松治疗的豚鼠(在30天内每天28毫克可的松/千克体重)的巨噬细胞。此外,还研究了诺卡菌对体外腹膜洗液中细胞的细胞毒性作用,以及正常血清、免疫血清和抗巨噬细胞血清对吞噬作用和细胞内增殖的影响。从腹膜洗液中获得的细胞中,巨噬细胞最常被吞噬,白细胞被吞噬的程度较低。正常巨噬细胞的吞噬率为14.56%(图1),两阶段免疫豚鼠的巨噬细胞为18.21%(图2),可的松治疗豚鼠的巨噬细胞为12.48%。细胞内观察显示,暴露3分钟后可见被吞噬的细菌。诺卡菌细胞内增殖的过程见图3。三组巨噬细胞的吞噬指数均缓慢增加;免疫巨噬细胞增加最少(8小时后为1.30)。可的松治疗豚鼠的巨噬细胞吞噬指数最高(8小时后为2.02)。暴露8小时内,诺卡菌的细丝穿过吞噬细胞的细胞膜(图4A、4B)。图5显示了诺卡菌对细胞的细胞致病作用过程。1小时后,死亡细胞数量从0.30%增加到1.9 - 3.8%;4小时后达到8.15 - 9.80%;8小时后为10.1 - 14.80%。可的松治疗豚鼠的细胞中观察到的数值最高(14.80%)。加入正常血清(吞噬时间60分钟)后,正常腹膜巨噬细胞的吞噬率为13.30%;免疫血清刺激吞噬作用(16.21%);抗巨噬细胞血清显著降低吞噬作用(4.10%)。加入正常血清和免疫血清后,腹膜巨噬细胞的吞噬指数增加,8小时内达到1.20 - 1.58;差异无统计学意义(图6)。抗巨噬细胞血清处理的腹膜巨噬细胞增殖显著增加(暴露8小时后为2.07)。正常豚鼠血清和免疫豚鼠血清均未影响诺卡菌对腹膜洗液细胞的细胞致病作用(图7)。抗巨噬细胞血清对巨噬细胞有快速且显著的作用(暴露2小时后78.23%的细胞死亡,3小时后100%死亡)。