Onyekaba C O, Harty J T, Plagemann P G
Department of Microbiology, Medical School, University of Minnesota, Minneapolis 55455.
Virus Res. 1989 Dec;14(4):327-38. doi: 10.1016/0168-1702(89)90025-7.
Indirect fluorescent antibody staining was used to examine the replication of lactate dehydrogenase-elevating virus (LDV) in primary cultures of peritoneal macrophages from BALB/c mice of different ages. Up to 80% of the total peritoneal macrophages from 1-2-week-old mice were susceptible to productive infection by LDV, though only 1-2% of the cells expressed detectable levels of IA antigen. The proportion of LDV-permissive peritoneal macrophages progressively decreased to 5-15% between 2 and 5 weeks of age of the mice. Macrophages from 9-day-old mice, when cultured in the presence of L cell conditioned medium, retained undiminished LDV permissiveness for at least 10 days in culture. The maximum proportion of LDV antigen-positive cells was detected between 8-10 h post infection of macrophages cultured from both 1-2-week-old and adult mice, concomitant with maximum LDV RNA synthesis. The LDV antigen positive macrophages disappeared between 12 and 48 h post infection. In cultures of macrophages from 9-10-day-old mice, the loss of infected cells was clearly due to cell killing, proving unequivocally that LDV replication is cytocidal. Disintegration of LDV-infected macrophages or phagocytosis of killed macrophages by surviving macrophages must be very sudden and complete since infected cells disappeared without the appearance of trypan blue-stainable cells in the culture. Ten cell lines established from macrophages of 2, 9, and 10-day-old mice all contained a small proportion of LDV-permissive cells (1-4%). Individual clones of one of the lines contained a similar small proportion of LDV-permissive cells.
采用间接荧光抗体染色法检测乳酸脱氢酶升高病毒(LDV)在不同年龄BALB/c小鼠腹腔巨噬细胞原代培养物中的复制情况。1 - 2周龄小鼠的腹腔巨噬细胞中,高达80%易受LDV的增殖性感染,尽管只有1 - 2%的细胞表达可检测水平的IA抗原。在小鼠2至5周龄期间,对LDV敏感的腹腔巨噬细胞比例逐渐降至5 - 15%。9日龄小鼠的巨噬细胞在L细胞条件培养基存在下培养时,在培养中至少10天保持对LDV的敏感性不变。在1 - 2周龄和成年小鼠培养的巨噬细胞感染后8 - 10小时检测到LDV抗原阳性细胞的最大比例,同时伴随最大量的LDV RNA合成。感染后12至48小时,LDV抗原阳性巨噬细胞消失。在9 - 10日龄小鼠巨噬细胞培养物中,感染细胞的损失显然是由于细胞杀伤,明确证明LDV复制具有细胞毒性。LDV感染的巨噬细胞解体或存活巨噬细胞对死亡巨噬细胞的吞噬作用一定非常突然且彻底,因为感染细胞消失时,培养物中未出现台盼蓝可染细胞。从2、9和10日龄小鼠巨噬细胞建立的10个细胞系均含有一小部分对LDV敏感的细胞(1 - 4%)。其中一个细胞系的单个克隆含有相似比例的对LDV敏感的细胞。