Biwei Cao1,2,3, Tao Wen1,2,3, Meng Wei1,2,3, Yuan Xiong1,2,3, Wan Liu1,2,3, Li Zhu1,2,3, Jing Zhou4. 1. Department of Tuina and Rehabilitation Medicine, Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan, 430061, China. 2. Department of Tuina and Rehabilitation Medicine, Affiliated Hospital of Hubei University of Chinese Medicine, Wuhan, 430061, China. 3. Department of Tuina and Rehabilitation Medicine, Hubei Province Academy of Traditional Chinese Medicine, Wuhan, 430074, China. 4. First Clinical Medical College, Hubei University of Chinese Medicine, Wuhan, 430061, China. zhoujing@hbhtcm.com.
Abstract
BACKGROUND: Vitamin D binding protein (VDBP) is an essential plasma carrier protein, which plays possible roles in reproductive health, disease and so on. However, the effects of VDBP on immunity have not been fully studied and the pertinent literatures remain very limited. OBJECTIVE: In this study, we introduced the exogenous VDBP into DC2.4 and established a stable DC2.4/VDBP cell line to explore the role of this gene in immunity. METHODS: Dendritic cells (DCs), as the most effective antigen presenting cells (APC) found so far, are directly involved in regulating some innate immunity. In order to evaluate the biological role of VDBP in DCs, we stably overexpressed VDBP in DCs, and conducted Cell Counting Kit‑8 (CCK-8 kit) and flow cytometry to detect changes in cell function. CCK-8 kit was used to monitor the viability of DCs after gene overexpression, and flow cytometry was used to detect changes in cell cycle distribution and apoptosis. Subsequently, in order to reveal the mechanism of VDBP regulating DCs, we adopted RNA sequencing (RNA-seq). RESULTS: CCK-8 results revealed VDBP successfully inhibited viability of DCs. Besides, we found that overexpression of this gene greatly promoted apoptosis and obviously altered the cell cycle distribution of DCs in G1 and G2 phases. Moreover, RNA-seq was carried out and 151 differently expression genes (DEGs) were obtained. In addition, gene differential expression analysis showed that most of them were uniformly enriched in immunity-related pathways. CONCLUSION: These results indicated that VDBP greatly repressed proliferation, facilitated apoptosis and changed cell cycle in DCs via altering the expression levels of gene associated with their cellular immunity.
BACKGROUND: Vitamin D binding protein (VDBP) is an essential plasma carrier protein, which plays possible roles in reproductive health, disease and so on. However, the effects of VDBP on immunity have not been fully studied and the pertinent literatures remain very limited. OBJECTIVE: In this study, we introduced the exogenous VDBP into DC2.4 and established a stable DC2.4/VDBP cell line to explore the role of this gene in immunity. METHODS: Dendritic cells (DCs), as the most effective antigen presenting cells (APC) found so far, are directly involved in regulating some innate immunity. In order to evaluate the biological role of VDBP in DCs, we stably overexpressed VDBP in DCs, and conducted Cell Counting Kit‑8 (CCK-8 kit) and flow cytometry to detect changes in cell function. CCK-8 kit was used to monitor the viability of DCs after gene overexpression, and flow cytometry was used to detect changes in cell cycle distribution and apoptosis. Subsequently, in order to reveal the mechanism of VDBP regulating DCs, we adopted RNA sequencing (RNA-seq). RESULTS: CCK-8 results revealed VDBP successfully inhibited viability of DCs. Besides, we found that overexpression of this gene greatly promoted apoptosis and obviously altered the cell cycle distribution of DCs in G1 and G2 phases. Moreover, RNA-seq was carried out and 151 differently expression genes (DEGs) were obtained. In addition, gene differential expression analysis showed that most of them were uniformly enriched in immunity-related pathways. CONCLUSION: These results indicated that VDBP greatly repressed proliferation, facilitated apoptosis and changed cell cycle in DCs via altering the expression levels of gene associated with their cellular immunity.
Authors: Benny Dahl; Frank V Schiødt; Peter Ott; Frank Wians; William M Lee; Jody Balko; Grant E O'Keefe Journal: Crit Care Med Date: 2003-01 Impact factor: 7.598
Authors: C Sue Carter; William M Kenkel; Evan L MacLean; Steven R Wilson; Allison M Perkeybile; Jason R Yee; Craig F Ferris; Hossein P Nazarloo; Stephen W Porges; John M Davis; Jessica J Connelly; Marcy A Kingsbury Journal: Pharmacol Rev Date: 2020-10 Impact factor: 25.468