Granger D L, Perfect J R, Durack D T
J Immunol. 1986 Jul 15;137(2):693-701.
Peritoneal macrophages from Mycobacterium bovis- or Toxoplasma gondii-infected mice cultured in vitro in Dulbecco's medium containing 10% fetal bovine serum (FBS) and endotoxin stopped replication of Cryptococcus neoformans for 30 hr, whereas yeast cells cultured alone reproduced with a 3.0-hr doubling time. Without at least 5% FBS, macrophage fungistasis was poor. FBS without macrophages enhanced the growth rate of cryptococci. Macrophages preincubated in vitro for 24 hr without serum became fungistatic when challenged with cryptococci in medium with FBS but were not fungistatic without FBS. Macrophages preincubated in medium with FBS were never subsequently fungistatic. Dialyzed, heated (56 degrees C, 30 min), or delipidated FBS supported macrophage fungistasis, whereas FBS heated at 70 degrees C for 30 min did not. FBS contained no measurable opsonic activity for C. neoformans. Inclusion of endotoxin and/or murine IFN-gamma over wide concentration ranges did not substitute for FBS. Ultrafiltration estimation of FBS activity localized to 50 to 150 Kd. By gel filtration chromatography, FBS activity ran in the 25 to 100 Kd range. Dye-ligand affinity chromatography on Cibacron blue agarose gel dissociated the FBS activity from the albumin and lipoprotein fractions. Anion-exchange chromatography on DEAE-Sephacel revealed activity in the first fraction eluting at low ionic strength, pointing to a protein(s) with an isoelectric point toward neutral. Activated macrophages can prevent microbial replication within host tissues; the local environment is critical for fulfillment of this important physiologic function. These results point to a macromolecular factor(s) present in serum that is essential for full fungistatic capability of activated macrophages.
来自感染牛分枝杆菌或刚地弓形虫小鼠的腹腔巨噬细胞，在含有10%胎牛血清（FBS）和内毒素的杜尔贝科培养基中体外培养时，可使新型隐球菌的复制停止30小时，而单独培养的酵母细胞则以3.0小时的倍增时间繁殖。若FBS含量低于5%，巨噬细胞的抑真菌作用则较差。没有巨噬细胞时，FBS会提高隐球菌的生长速率。在无血清条件下体外预孵育24小时的巨噬细胞，当在含有FBS的培养基中受到隐球菌攻击时会变成抑真菌状态，但在没有FBS时则无抑真菌作用。在含有FBS的培养基中预孵育的巨噬细胞随后再也不会具有抑真菌作用。经过透析、加热（56℃，30分钟）或脱脂的FBS可支持巨噬细胞的抑真菌作用，而在70℃加热30分钟的FBS则不能。FBS对新型隐球菌没有可检测到的调理活性。在很宽的浓度范围内加入内毒素和/或鼠干扰素-γ并不能替代FBS。FBS活性的超滤估计定位于50至150千道尔顿。通过凝胶过滤色谱法，FBS活性在25至100千道尔顿范围内。在蓝色葡聚糖琼脂糖凝胶上进行染料配体亲和色谱法可将FBS活性与白蛋白和脂蛋白组分分离。在DEAE-葡聚糖凝胶上进行阴离子交换色谱法显示，在低离子强度下洗脱的第一部分中有活性，表明存在一种等电点趋向中性的蛋白质。活化的巨噬细胞可防止微生物在宿主组织内复制；局部环境对于实现这一重要生理功能至关重要。这些结果表明血清中存在一种大分子因子，它对于活化巨噬细胞的完全抑真菌能力至关重要。
