Department of Biology and South Texas Center for Emerging Infectious Diseases, University of Texas at San Antonio, San Antonio, TX 78249, USA.
Microb Pathog. 2011 Feb;50(2):100-8. doi: 10.1016/j.micpath.2010.11.006. Epub 2010 Dec 1.
We studied the effect of the presence of Coccidioides on the production of nitric oxide (NO) by primary macrophages previously activated by IFN-γ and LPS. The fungal cells were isolated from cultures of arthroconidia that had been incubated for 24 h in a medium that supported parasitic phase growth and were co-cultured with the macrophages. These live, first-generation parasitic cells of Coccidioides, referred to as spherule initials, suppressed NO production as well as iNOS mRNA expression by activated macrophages. Phagocytosis was not required for suppression of NO. We also showed that the culture supernatant of the spherule initials was capable of suppressing NO production, and that this activity was mediated by an as yet unidentified, secreted fungal factor(s). Heat-, paraformaldehyde- or X-ray-treated spherule initials did not show this inhibitory effect. To our surprise, macrophages obtained from iNOS-deficient mice revealed phagocytic activity and killing efficiency which were comparable to that of macrophages isolated from wild type C57BL/6 mice. Although the cultured fungal pathogen can suppress NO production, this oxidative product is apparently not essential for in vitro killing of Coccidioides by activated macrophages. Our results suggest that other unidentified fungicidal mechanisms exist against Coccidioides which are apparently independent of NO production.
我们研究了在 IFN-γ 和 LPS 预先激活的原代巨噬细胞中,粗球孢子菌的存在对一氧化氮(NO)产生的影响。从培养的关节孢子中分离出真菌细胞,这些关节孢子在支持寄生阶段生长的培养基中孵育了 24 小时,并与巨噬细胞共培养。这些活的、第一代的粗球孢子菌寄生细胞,称为小球体初始细胞,抑制了激活的巨噬细胞的 NO 产生和 iNOS mRNA 表达。吞噬作用对于抑制 NO 不是必需的。我们还表明,小球体初始细胞的培养上清液能够抑制 NO 的产生,并且这种活性是由一种尚未确定的、分泌的真菌因子介导的。热、多聚甲醛或 X 射线处理的小球体初始细胞没有显示这种抑制作用。令我们惊讶的是,从 iNOS 缺陷型小鼠中获得的巨噬细胞显示出吞噬活性和杀伤效率,与从野生型 C57BL/6 小鼠中分离的巨噬细胞相当。尽管培养的真菌病原体可以抑制 NO 的产生,但这种氧化产物对于激活的巨噬细胞体外杀伤粗球孢子菌显然不是必需的。我们的结果表明,针对粗球孢子菌存在其他未被识别的杀菌机制,显然与 NO 的产生无关。