Diamond Gill, Figgins Erika L, Gao Denny, Robinson Tanya O, Wilson Kenneth, Gabbard Jon D, Anderson Charles D, Batra Lalit, Sreenivasan Chithra, Zheng Jian, Do Uyen, Mulcahy Braden, Ryan Lisa K, McLaughlin Nathaniel, Owings Anna H, Dhaliwal Neha, Williams Haley B, Wichman T Grant, Galeas-Pena Michelle, Glover Sarah C
Department of Oral Immunology and Infectious Diseases, University of Louisville School of Dentistry, Louisville, KY, USA.
Center for Predictive Medicine for Biodefence and Emerging Infectious Diseases, University of Louisville, Louisville, KY, USA.
Sci Rep. 2024 Dec 30;14(1):31740. doi: 10.1038/s41598-024-82098-z.
The Paneth cell, a secretory cell of the small intestine, expresses numerous host defense proteins, and is hypothesized to play an important role in host defense against infection. However, studying gene expression in this cell requires invasive procedures. To test the hypothesis that we could observe Paneth cell-specific gene regulation from exfoliated cells in infectious conditions, we obtained stool samples from patients with COVID-19 and uninfected controls. Total mRNA was isolated, and Paneth cell-specific and non-specific gene expression was quantified by RT-PCR. Results revealed a significant decrease in mRNA levels from Paneth cell-specific genes, including DEFA5, DEFA6, PLA2G2A, PRSS2 and ITLN2 in SARS-CoV-2 positive patients compared with controls. Other gut markers, not specific to Paneth cells were unchanged. To validate this experimentally, we infected mice with SARS-CoV-2 and collected fecal pellets over the course of 7 days. We observed a similar time-dependent reduction in Paneth cell-specific transcripts, which correlates with histological changes in the gut. This is the first demonstration of quantification of Paneth cell-specific transcripts without invasive sampling. It also shows the coordinate regulation of these genes as a response to infection with SARS-CoV-2, possibly through viral pathogenesis, to increase infectivity in the gut.
潘氏细胞是小肠的一种分泌细胞，表达多种宿主防御蛋白，据推测在宿主抵御感染中发挥重要作用。然而，研究该细胞中的基因表达需要侵入性操作。为了验证我们能够在感染情况下从脱落细胞中观察到潘氏细胞特异性基因调控这一假设，我们从新冠肺炎患者和未感染的对照者那里获取了粪便样本。分离出总mRNA，并通过逆转录聚合酶链反应（RT-PCR）对潘氏细胞特异性和非特异性基因表达进行定量。结果显示，与对照组相比，新冠病毒阳性患者中潘氏细胞特异性基因（包括防御素A5、防御素A6、磷脂酶A2G2A、丝氨酸蛋白酶2和肠三叶因子2）的mRNA水平显著降低。其他非潘氏细胞特异性的肠道标志物则没有变化。为了通过实验验证这一点，我们用新冠病毒感染小鼠，并在7天内收集粪便颗粒。我们观察到潘氏细胞特异性转录本出现了类似的时间依赖性减少，这与肠道的组织学变化相关。这是首次在不进行侵入性采样的情况下对潘氏细胞特异性转录本进行定量。它还表明，这些基因作为对新冠病毒感染的一种反应，可能通过病毒致病机制进行协同调控，从而增加肠道中的感染性。
