Department of Cell Biology, Haffkine Institute for Training, Research & Testing, Mumbai, India.
Department of Virology & Immunology, Haffkine Institute for Training, Research & Testing, Mumbai, India.
Eur J Haematol. 2020 Mar;104(3):170-180. doi: 10.1111/ejh.13350. Epub 2019 Dec 26.
HIV-1-infected patients develop haematological disorders such as cytopenias. One possible explanation is the inhibition of haematopoiesis at the level of differentiation of CD34+ haematopoietic progenitor stem cells. Based on our previous studies, we hypothesised that there may be viral encoded, or host cellular factors which participate in the process of inhibition of haematopoiesis.
Virus-depleted media from infected CD4+ T cells was prepared by filtration and added to CD34+ cell differentiation semisolid medium. We have also used the virus-depleted media to isolate host/viral factors including miRNA. Isolated miRNAs were screened for their haematopoietic inhibitory function using the miRNA mining approach.
Addition of virus-depleted media caused a 40% inhibition of differentiation of CD34+ cells into myeloid and erythroid colony formation. Real-time RT-PCR showed miR-15a and miR-24 from both pIndie-C1 and pNL4.3 HIV-1-infected cells showed a significant differential expression when compared to control media.
In this study, we have identified two miRNAs, miR-15a and miR-24 secreted from purified HIV-1-infected CD4+ T cells that inhibited CD34+ haematopoietic progenitor stem cell differentiation into myeloid and erythroid colonies in vitro.
HIV-1 感染患者会出现血液学紊乱，例如细胞减少症。一种可能的解释是 CD34+造血祖细胞干细胞分化水平的造血抑制。基于我们之前的研究，我们假设可能存在参与造血抑制过程的病毒编码或宿主细胞因子。
通过过滤制备感染的 CD4+T 细胞的病毒耗尽培养基，并将其添加到 CD34+细胞分化半固体培养基中。我们还使用病毒耗尽的培养基来分离宿主/病毒因子，包括 miRNA。使用 miRNA 挖掘方法筛选分离的 miRNA 以评估其造血抑制功能。
添加病毒耗尽的培养基会导致 CD34+细胞向髓系和红细胞集落形成的分化减少 40%。实时 RT-PCR 显示，与对照培养基相比，来自 pIndie-C1 和 pNL4.3 HIV-1 感染细胞的 miR-15a 和 miR-24 的表达明显不同。
在这项研究中，我们鉴定了两种 miRNA，即来自纯化的 HIV-1 感染的 CD4+T 细胞的 miR-15a 和 miR-24，它们在体外抑制 CD34+造血祖细胞干细胞向髓系和红细胞集落的分化。
