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AAV-Vectored Fms-Related Tyrosine Kinase 3 Ligand Inhibits CD34+ Progenitor Cell Engraftment in Humanized Mice

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Abstract

Humanized mice have become useful animal models for HIV/AIDS. Since NOD.Cg-Prkdc scid Il2rgtm1Wjl/SzJ (NSG) mice allow the engraftment of primary human immune cells, we aim to determine the role of human Fms-related tyrosine kinase 3 ligand (hFlt3L), a major growth factor for dendritic cells (DCs), in regulating the differentiation of cord blood-derived CD34+ progenitor cells in this murine species. Soluble recombinant hFlt3L protein and AAV-vectored hFlt3L were administrated before or after human CD34+ progenitor cell transplantation, respectively. We then measured the peripheral levels of hFlt3L by ELISA. Meantime, reconstituted human immune cells were analyzed by flow cytometry over time. We found that without hFlt3L there were significantly increased types of human immune cells in NSG-huCD34 compared with NSG-huPBL mice but the frequency of human DCs remains low. Transient treatment with recombinant hFlt3L expanded human conventional CD1c+ and CD141+ DCs as well as plasmacytoid DCs in humanized NSG-huCD34 mice. Surprisingly, however, the prolonged in vivo expression of AAV-vectored hFlt3L resulted in significant suppression of total human CD34+ cell engraftment and differentiation. The suppression occurred within 2 weeks when AAV-vectored hFlt3L was administered either before or after the transplantation of CD34+ progenitor cells, which was likely associated with the induction of murine myeloid-derived immune suppressive cells and reactive oxygen species in NSG-huCD34 mice. Since chronic  HIV-1 patients displayed significantly high levels of hFlt3L expression, our findings may have implication to explore the role of prolonged hFlt3L in regulating  the differentiation of human CD34+ progenitor cells in both NSG-huCD34 mice and infected people.

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Acknowledgments

ZC and LL designed the study, analyzed data and wrote the manuscript. LL, XT, XH, JL did the experiments. HW provided technical supports. The authors would like to thank Hong Kong Research Grant Council TRS T11-706/18-N, HKU5/CRF/13G, the San-Ming Project of Medicine in Shenzhen and the High Level Hospital-Summit Program in Guangdong for financial supports.

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Authors and Affiliations

  1. AIDS Institute and Department of Microbiology, State Key Laboratory of Emerging Infectious Diseases, Li Ka Shing Faculty of Medicine, The University of Hong Kong, L5-45, Laboratory Block, 21 Sassoon Road, Pokfulam, Hong Kong Special Administrative Region, People’s Republic of China

    Lijun Ling, Xiuyan Huang, Jingjing Li & Zhiwei Chen

  2. University of Hong Kong AIDS Institute-Shenzhen Research Laboratory, Guangdong Key Laboratory of Emerging Infectious Diseases and Shenzhen Key Laboratory of Infection and Immunity, Shenzhen Third People’s Hospital, Shenzhen, People’s Republic of China

    Lijun Ling, Xian Tang, Hui Wang & Zhiwei Chen

  3. University of Hong Kong-Shenzhen Hospital, Shenzhen, People’s Republic of China

    Zhiwei Chen

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  1. Lijun Ling
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Correspondence to Zhiwei Chen.

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Ling, L., Tang, X., Huang, X. et al. AAV-Vectored Fms-Related Tyrosine Kinase 3 Ligand Inhibits CD34+ Progenitor Cell Engraftment in Humanized Mice. J Neuroimmune Pharmacol 13, 541–550 (2018). https://doi.org/10.1007/s11481-018-9819-0

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