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首页> 外文期刊>Infection and immunity >Murine peritoneal macrophages activated by the mycobacterial 65-kilodalton heat shock protein express enhanced microbicidal activity in vitro.
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Murine peritoneal macrophages activated by the mycobacterial 65-kilodalton heat shock protein express enhanced microbicidal activity in vitro.

机译：由分枝杆菌65-千洛酮热休克蛋白激活的小鼠腹膜巨噬细胞在体外表达增强的杀菌活性。

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After an intraperitoneal (i.p.) injection of purified protein derivative, peritoneal macrophages from mice infected with Mycobacterium bovis bacillus Calmette-Guérin (BCG) show an enhanced respiratory burst, inhibit the intracellular proliferation of Toxoplasma gondii, and kill Listeria monocytogenes more efficiently than peritoneal macrophages from normal mice. One of the immunodominant antigens of Mycobacterium spp. is the 65-kDa heat shock protein (Hsp 65), and in the present study, we determined whether injection of this protein into mice leads to activation of their peritoneal macrophages. After an i.p. injection of Hsp 65, peritoneal macrophages from BCG-infected CBA/J mice also released more H2O2, inhibited the proliferation of T. gondii, and killed L. monocytogenes faster than peritoneal macrophages from normal mice, although Hsp 65 was less effective than purified protein derivative. When normal mice were injected with Hsp 65 suspended in saline after a booster injection with Hsp 65, their macrophages did not display enhanced antimicrobial activity, indicating that an adjuvant was required for a cellular immune response against Hsp 65. In the present study, the adjuvant dimethyl dioctadecylammonium bromide (DDA) was preferred because it contains no endotoxin or mycobacterial antigens and because it has been reported that DDA does not induce the production of gamma interferon. Peritoneal macrophages from C57BL/6 and CBA/J mice that had received a subcutaneous injection of Hsp 65 suspended in DDA followed by an i.p. booster injection of Hsp 65 suspended in saline were activated, as indicated by the enhanced production of H2O2, inhibition of the intracellular proliferation of T. gondii, and increased rate of intracellular killing of L. monocytogenes in vitro relative to that by resident peritoneal macrophages and peritoneal macrophages obtained from mice that had received ovalbumin instead of Hsp 65. The rate of phagocytosis of L. monocytogenes was not affected by Hsp 65 treatment. Despite the in vitro expression of enhanced microbicidal activity of peritoneal macrophages, no difference in the growth of L. monocytogenes in the liver and spleen between Hsp 65-treated and control mice was found.
机译：腹膜内（ip）注射纯化的蛋白衍生物后，感染牛分枝杆菌Calmette-Guérin（BCG）的小鼠的腹腔巨噬细胞显示出增强的呼吸爆发，抑制弓形虫的细胞内增殖，并且比腹膜巨噬细胞更有效地杀死单核细胞增生性李斯特菌来自正常小鼠。分枝杆菌属的免疫优势抗原之一。是65 kDa的热休克蛋白（Hsp 65），在本研究中，我们确定了将该蛋白注射入小鼠是否会导致其腹膜巨噬细胞活化。在ip之后注射Hsp 65后，来自BCG感染的CBA / J小鼠的腹膜巨噬细胞也释放出更多的H2O2，抑制弓形虫的增殖，并且杀死单核细胞增生李斯特菌的速度比正常小鼠的腹膜巨噬细胞要快，尽管Hsp 65的效果不如纯化的蛋白衍生物。当正常小鼠在注射Hsp 65的加强剂量后注射悬浮在盐水中的Hsp 65时，它们的巨噬细胞没有显示出增强的抗微生物活性，这表明佐剂是针对Hsp 65的细胞免疫反应所必需的。在本研究中，佐剂优选二甲基二十八烷基溴化铵（DDA），因为它不包含内毒素或分枝杆菌抗原，并且据报道，DDA不会诱导γ干扰素的产生。来自C57BL / 6和CBA / J小鼠的腹膜巨噬细胞，这些小鼠接受了DDA皮下注射的Hsp 65的皮下注射，随后腹膜内注射。相对于常驻腹膜巨噬细胞和巨噬细胞，H2O2产生增加，抑制弓形虫细胞内增殖以及体外对单核细胞增生李斯特氏菌的细胞内杀灭率提高表明，悬浮在盐水中的Hsp 65的加强注射被激活。从接受卵清蛋白而不是Hsp 65的小鼠获得的腹膜巨噬细胞。单核细胞增生李斯特氏菌的吞噬率不受Hsp 65处理的影响。尽管在体外表达了增强的腹膜巨噬细胞杀微生物活性，但是在经Hsp 65处理的小鼠和对照小鼠之间，未发现肝和脾中单核细胞增生李斯特氏菌的生长差异。

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