Vats Amrita, Ho Tzu-Chuan, Puc Irwin, Chen Yi-Ju, Chang Chiung-Hsin, Chien Yu-Wen, Perng Guey-Chuen
Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
Department of Microbiology and Immunology, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
Heliyon. 2021 Apr 20;7(4):e06785. doi: 10.1016/j.heliyon.2021.e06785. eCollection 2021 Apr.
Recent studies have shown that dengue virus (DENV) can efficiently infect bone marrow hematopoietic stem cells (HSCs) as well as the placenta of pregnant women. Although mother-to-infant vertical transmission of DENV through the placenta has been well documented, the evidence of cell-associated vertical transmission is still unknown. Whether DENV can infect umbilical cord blood (UCB) cells before reaching the fetus remains to be explored. Here, we proposed that human UCB cells were permissive to the DENV infection and DENV infected CD133+ and CD34+ HSCs are reservoir of the virus that could be reactivated upon re-culturing in suitable cells.
Human UCB cells were freshly obtained and subjected to DENV infection. Multicolor flow cytometry (MFCM) was used to demonstrate the phenotypes of the infected HSC populations. Immunofluorescence analysis (IFA) and T-distributed Stochastic Neighbor Embedding (t-SNE) were used to show the association of the DENV antigen, non-structural protein1 (NS1) with HSCs.
UCB cells were highly permissive to DENV infection. DENV altered the phenotype of the infected HSC population, increased the expression of HSCs, and affected the balance of transcription factors (TFs, GATA1/2/3). IFA revealed the association of the DENV antigen, non-structural protein1 (NS1), with CD34 and CD133 cells. T-distributed Stochastic Neighbor Embedding (t-SNE) analysis revealed heterogeneity in the distribution of CD133NS1, and CD34 NS1 cells. DENV particles were recovered from CD133 and CD34 cells even when virus production in the supernatant was negligible.
We predict that infection of CD133+ and CD34+ cells in the UCB serve as reservoirs for the amplification of DENV in UCB prior to the virus reaching the fetus and facilitate vertical transmission.
近期研究表明，登革病毒（DENV）可有效感染骨髓造血干细胞（HSCs）以及孕妇的胎盘。尽管已有充分文献记载DENV通过胎盘进行母婴垂直传播，但细胞相关垂直传播的证据仍不明确。DENV在到达胎儿之前是否能够感染脐带血（UCB）细胞仍有待探索。在此，我们提出人UCB细胞对DENV感染具有易感性，且感染DENV的CD133+和CD34+造血干细胞是该病毒的储存库，在合适的细胞中重新培养时可被重新激活。
新鲜获取人UCB细胞并使其感染DENV。采用多色流式细胞术（MFCM）来展示被感染造血干细胞群体的表型。免疫荧光分析（IFA）和T分布随机邻域嵌入（t-SNE）用于显示DENV抗原、非结构蛋白1（NS1）与造血干细胞的关联。
UCB细胞对DENV感染具有高度易感性。DENV改变了被感染造血干细胞群体的表型，增加了造血干细胞的表达，并影响转录因子（TFs，GATA1/2/3）的平衡。IFA显示DENV抗原、非结构蛋白1（NS1）与CD34和CD133细胞有关联。T分布随机邻域嵌入（t-SNE）分析揭示了CD133NS1和CD34 NS1细胞分布的异质性。即使上清液中的病毒产量可忽略不计，仍可从CD133和CD34细胞中回收DENV颗粒。
我们预测，UCB中CD133+和CD34+细胞的感染在病毒到达胎儿之前可作为UCB中DENV扩增的储存库，并促进垂直传播。
