Hainan Key Laboratory of Tropical Animal Reproduction & Breeding and Epidemic Disease Research, Animal Genetic Engineering Key Lab of Haikou, College of Animal Science and Technology, Hainan University, Haikou 570228, China.
Genes (Basel). 2022 Sep 3;13(9):1586. doi: 10.3390/genes13091586.
() is an opportunistic pathogen that is common in livestock and poultry and leads to massive economic losses in the animal husbandry sector. In this study, we challenged mice with strain HN02 by intraperitoneal injection and collected spleens to measure bacterial loads. We also performed histopathological analysis by hematoxylin and eosin (H&E) staining. Then we used RNA-sequencing (RNA-seq) to detect the mRNA expression levels in the mouse spleen and quantitative real-time PCR (qRT-PCR) to verify the sequencing data. Finally, we examined the effect of HN02 on anti-inflammatory cytokine interleukin-10 (IL-10) protein expression in the spleen through immunohistochemical analysis. The results showed that compared to those in the control group, the mouse spleens in the challenge group had lesions, and the average bacteria loads was (3.07 ± 1.09) × 10 CFU (colony-forming unit)/g. The RNA-seq results determined 3653 differentially expressed genes (DEGs), and the qRT-PCR analysis revealed immune-related genes consistent with the expression trend in the sequencing data. The number and area of IL-10 positive cells substantially increased to resist inflammation in the challenge group. In conclusion, we analyzed the spleens of mice infected with from multiple perspectives, and our findings lay a foundation for subsequent studies on the mechanism of pathogen-host interactions.
是一种机会性病原体，常见于家畜和家禽，导致畜牧业部门遭受巨大的经济损失。在本研究中，我们通过腹腔注射挑战小鼠感染 株 HN02，并收集脾脏来测量细菌载量。我们还通过苏木精和伊红（H&E）染色进行组织病理学分析。然后，我们使用 RNA 测序（RNA-seq）检测小鼠脾脏中的 mRNA 表达水平，并通过定量实时 PCR（qRT-PCR）验证测序数据。最后，我们通过免疫组织化学分析检测了 HN02 对脾脏抗炎细胞因子白细胞介素 10（IL-10）蛋白表达的影响。结果表明，与对照组相比，感染组小鼠的脾脏出现病变，平均细菌载量为（3.07±1.09）×10 CFU（菌落形成单位）/g。RNA-seq 结果确定了 3653 个差异表达基因（DEGs），qRT-PCR 分析显示免疫相关基因与测序数据的表达趋势一致。在感染组中，IL-10 阳性细胞的数量和面积显著增加，以抵抗炎症。总之，我们从多个角度分析了感染 的小鼠的脾脏，我们的研究结果为后续研究病原体-宿主相互作用的机制奠定了基础。
