Department of Molecular Biology and Microbiology, Tufts University School of Medicine and Howard Hughes Medical Institute, Boston, Massachusetts, USA.
Infect Immun. 2014 Jun;82(6):2434-47. doi: 10.1128/IAI.00054-14. Epub 2014 Mar 31.
Vibrio cholerae is the causative agent of the acute diarrheal disease of cholera. Innate immune responses to V. cholerae are not a major cause of cholera pathology, which is characterized by severe, watery diarrhea induced by the action of cholera toxin. Innate responses may, however, contribute to resolution of infection and must be required to initiate adaptive responses after natural infection and oral vaccination. Here we investigated whether a well-established infant mouse model of cholera can be used to observe an innate immune response. We also used a vaccination model in which immunized dams protect their pups from infection through breast milk antibodies to investigate innate immune responses after V. cholerae infection for pups suckled by an immune dam. At the peak of infection, we observed neutrophil recruitment accompanied by induction of KC, macrophage inflammatory protein 2 (MIP-2), NOS-2, interleukin-6 (IL-6), and IL-17a. Pups suckled by an immunized dam did not mount this response. Accessory toxins RtxA and HlyA played no discernible role in neutrophil recruitment in a wild-type background. The innate response to V. cholerae deleted for cholera toxin-encoding phage (CTX) and part of rtxA was significantly reduced, suggesting a role for CTX-carried genes or for RtxA in the absence of cholera toxin (CTX). Two extracellular V. cholerae DNases were not required for neutrophil recruitment, but DNase-deficient V. cholerae caused more clouds of DNA in the intestinal lumen, which appeared to be neutrophil extracellular traps (NETs), suggesting that V. cholerae DNases combat NETs. Thus, the infant mouse model has hitherto unrecognized utility for interrogating innate responses to V. cholerae infection.
霍乱弧菌是引起霍乱这种急性腹泻病的病原体。先天免疫反应不是霍乱病理的主要原因,其特征是霍乱毒素作用引起严重的水样腹泻。然而,先天反应可能有助于感染的解决,并且在自然感染和口服疫苗接种后必须启动适应性反应。在这里,我们研究了一个成熟的婴儿小鼠霍乱模型是否可用于观察先天免疫反应。我们还使用了一种疫苗接种模型,其中免疫母鼠通过乳腺抗体保护其幼崽免受感染,以研究感染霍乱弧菌后,通过免疫母鼠哺乳的幼崽的先天免疫反应。在感染高峰时,我们观察到中性粒细胞募集伴随着 KC、巨噬细胞炎症蛋白 2 (MIP-2)、NOS-2、白细胞介素 6 (IL-6) 和 IL-17a 的诱导。由免疫母鼠哺乳的幼崽没有产生这种反应。在野生型背景下,辅助毒素 RtxA 和 HlyA 在中性粒细胞募集中没有明显作用。缺失编码霍乱毒素噬菌体 (CTX) 和 rtxA 部分的霍乱弧菌的先天反应明显减少,表明 CTX 携带的基因或在没有霍乱毒素 (CTX) 的情况下,RtxA 发挥作用。两种细胞外霍乱弧菌 DNase 对于中性粒细胞募集不是必需的,但是 DNase 缺陷型霍乱弧菌在肠腔中引起更多的 DNA 云,这些 DNA 云似乎是中性粒细胞细胞外陷阱 (NETs),表明霍乱弧菌 DNase 对抗 NETs。因此,婴儿小鼠模型迄今在研究对霍乱弧菌感染的先天反应方面具有尚未被认识到的用途。