Iizawa Y, Brown J F, Czuprynski C J
Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison 53706.
Infect Immun. 1992 Oct;60(10):4068-73. doi: 10.1128/iai.60.10.4068-4073.1992.
Protective immunity first becomes evident at 3 to 4 days after inoculation of mice with a sublethal dose of Listeria monocytogenes. Recent evidence suggests that production of gamma interferon (IFN-gamma) occurs earlier (within the first 24 h of infection). The purpose of this study was to define better the sequence of cytokine mRNA expression during the early stages of L. monocytogenes infection. Cytokine mRNA expression was detected by polymerase chain reaction-assisted amplification of RNA extracted from the spleen cells of individual mice euthanized at 0.5 to 120 h after L. monocytogenes challenge. By using this method, mRNAs for tumor necrosis factor alpha, interleukin-1 alpha (IL-1 alpha), IL-2, IL-4, IL-5, and IFN-gamma were detected in RNA from the spleen cells of uninfected mice. The intensity of the bands for IFN-gamma, however, was increased greatly at 16 h after intravenous injection of 5 x 10(4) CFU (nearly 1 50% lethal dose) of L. monocytogenes. IL-6 and granulocyte-macrophage colony-stimulating factor mRNAs were not detected in spleen cell RNA from uninfected mice but were induced within 30 and 60 min, respectively, after inoculation with L. monocytogenes. Increased amounts of mRNAs for IFN-gamma, IL-6, and granulocyte-macrophage colony-stimulating factor were detected after injection of viable, but not killed, L. monocytogenes. IL-3 mRNA was not detected at any time in RNA extracted from the spleen cells of uninfected or L. monocytogenes infected mice. These results suggest that infection with L. monocytogenes elicits a detectable cytokine mRNA response within the first few hours of infection.
用亚致死剂量的单核细胞增生李斯特菌接种小鼠后,保护性免疫在3至4天首次明显显现。最近的证据表明,γ干扰素(IFN-γ)的产生更早(在感染的最初24小时内)。本研究的目的是更好地确定单核细胞增生李斯特菌感染早期细胞因子mRNA表达的顺序。通过聚合酶链反应辅助扩增从单核细胞增生李斯特菌攻击后0.5至120小时安乐死的个体小鼠脾脏细胞中提取的RNA,检测细胞因子mRNA表达。通过使用这种方法,在未感染小鼠的脾脏细胞RNA中检测到肿瘤坏死因子α、白细胞介素-1α(IL-1α)、IL-2、IL-4、IL-5和IFN-γ的mRNA。然而,在静脉注射5×10⁴CFU(接近1/50致死剂量)的单核细胞增生李斯特菌后16小时,IFN-γ条带的强度大大增加。未感染小鼠的脾脏细胞RNA中未检测到IL-6和粒细胞-巨噬细胞集落刺激因子mRNA,但在接种单核细胞增生李斯特菌后分别在30和60分钟内被诱导。注射活的而非灭活的单核细胞增生李斯特菌后,检测到IFN-γ、IL-6和粒细胞-巨噬细胞集落刺激因子的mRNA量增加。在未感染或单核细胞增生李斯特菌感染小鼠的脾脏细胞提取的RNA中,任何时候都未检测到IL-3 mRNA。这些结果表明,单核细胞增生李斯特菌感染在感染的最初几个小时内引发了可检测到的细胞因子mRNA反应。