Transcriptomic Analysis Reveal the Responses of Dendritic Cells to VDBP

Overview

Journal Genes Genomics

Specialty Genetics

Date 2022 Mar 12

PMID 35278207

Authors

Biwei Cao

Tao Wen

Meng Wei

Yuan Xiong

Wan Liu

Li Zhu

Jing Zhou

Affiliations

Soon will be listed here.

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.

Citing Articles

Hypovitaminosis D in persons with Down syndrome and autism spectrum disorder.

Boyd N, Nguyen J, Khoshnood M, Jiang T, Nguyen L, Mendez L J Neurodev Disord. 2023; 15(1):35.

PMID: 37880588 PMC: 10599027. DOI: 10.1186/s11689-023-09503-y.

Transcriptome Analysis Reveals the Immunoregulatory Activity of Rice Seed-Derived Peptide PEP1 on Dendritic Cells.

Qu T, He S, Wu Y, Wang Y, Ni C, Wen S Molecules. 2023; 28(13).

PMID: 37446885 PMC: 10343632. DOI: 10.3390/molecules28135224.

References

Akiba T, Morikawa T, Odaka M, Nakada T, Kamiya N, Yamashita M . Vitamin D Supplementation and Survival of Patients with Non-small Cell Lung Cancer: A Randomized, Double-Blind, Placebo-Controlled Trial. Clin Cancer Res. 2018; 24(17):4089-4097. DOI: 10.1158/1078-0432.CCR-18-0483. View

Bag A, Saha S, Sundar S, Saha B, Chakrabarti A, Mandal C . Comparative proteomics and glycoproteomics of plasma proteins in Indian visceral leishmaniasis. Proteome Sci. 2014; 12(1):48. PMC: 4179796. DOI: 10.1186/s12953-014-0048-z. View

Behrouz G, Farzaneh G, Leila J, Jaleh Z, Eskandar K . Presence of auto-antibody against two placental proteins, annexin A1 and vitamin D binding protein, in sera of women with pre-eclampsia. J Reprod Immunol. 2013; 99(1-2):10-6. DOI: 10.1016/j.jri.2013.04.007. View

Berger D, Winter M, Beger H . Influence of human transferrin and group-specific protein on endotoxicity in vitro. Clin Chim Acta. 1990; 189(1):1-5. DOI: 10.1016/0009-8981(90)90227-j. View

Binder R, Kress A, Kan G, Herrmann K, Kirschfink M . Neutrophil priming by cytokines and vitamin D binding protein (Gc-globulin): impact on C5a-mediated chemotaxis, degranulation and respiratory burst. Mol Immunol. 2000; 36(13-14):885-92. DOI: 10.1016/s0161-5890(99)00110-8. View

Braun A, Kofler A, Morawietz S, CLEVE H . Sequence and organization of the human vitamin D-binding protein gene. Biochim Biophys Acta. 1993; 1216(3):385-94. DOI: 10.1016/0167-4781(93)90005-x. View

Carter C, Kenkel W, MacLean E, Wilson S, Perkeybile A, Yee J . Is Oxytocin "Nature's Medicine"?. Pharmacol Rev. 2020; 72(4):829-861. PMC: 7495339. DOI: 10.1124/pr.120.019398. View

Cavanagh L, Weninger W . Dendritic cell behaviour in vivo: lessons learned from intravital two-photon microscopy. Immunol Cell Biol. 2008; 86(5):428-38. DOI: 10.1038/icb.2008.25. View

Cribbs A, Kennedy A, Gregory B, Brennan F . Simplified production and concentration of lentiviral vectors to achieve high transduction in primary human T cells. BMC Biotechnol. 2013; 13:98. PMC: 3830501. DOI: 10.1186/1472-6750-13-98. View

10.

Dahl B, Schiodt F, Ott P, Wians F, Lee W, Balko J . Plasma concentration of Gc-globulin is associated with organ dysfunction and sepsis after injury. Crit Care Med. 2003; 31(1):152-6. DOI: 10.1097/00003246-200301000-00024. View

11.

de Menezes S, Kelly J, Wolfe R, Farrugia H, Mar V . The increasing use of shave biopsy for diagnosing invasive melanoma in Australia. Med J Aust. 2019; 211(5):213-218. DOI: 10.5694/mja2.50289. View

12.

Delanghe J, Speeckaert R, Speeckaert M . Behind the scenes of vitamin D binding protein: more than vitamin D binding. Best Pract Res Clin Endocrinol Metab. 2015; 29(5):773-86. DOI: 10.1016/j.beem.2015.06.006. View

13.

Diaz-Ochoa V, Jellbauer S, Klaus S, Raffatellu M . Transition metal ions at the crossroads of mucosal immunity and microbial pathogenesis. Front Cell Infect Microbiol. 2014; 4:2. PMC: 3900919. DOI: 10.3389/fcimb.2014.00002. View

14.

DiMartino S, Shah A, Trujillo G, Kew R . Elastase controls the binding of the vitamin D-binding protein (Gc-globulin) to neutrophils: a potential role in the regulation of C5a co-chemotactic activity. J Immunol. 2001; 166(4):2688-94. DOI: 10.4049/jimmunol.166.4.2688. View

15.

Dueland S, Nenseter M, Drevon C . Uptake and degradation of filamentous actin and vitamin D-binding protein in the rat. Biochem J. 1991; 274 ( Pt 1):237-41. PMC: 1149943. DOI: 10.1042/bj2740237. View

16.

Freire-Garabal M, Balboa J, Nunez M, Castano M, Llovo J, Belmonte A . Effects of amphetamine on T-cell immune response in mice. Life Sci. 1991; 49(16):PL107-12. DOI: 10.1016/0024-3205(91)90570-2. View

17.

Goodwin K, Nelson C . Generating tissue topology through remodeling of cell-cell adhesions. Exp Cell Res. 2017; 358(1):45-51. PMC: 5544595. DOI: 10.1016/j.yexcr.2017.03.016. View

18.

Govender P, Keyes C, Hankinson E, OHara C, Sanchorawala V, Berk J . Transbronchial biopsies safely diagnose amyloid lung disease. Amyloid. 2017; 24(1):37-41. PMC: 6014610. DOI: 10.1080/13506129.2017.1301917. View

19.

Guha C, Osawa M, Werner P, Galbraith R, Paddock G . Regulation of human Gc (vitamin D--binding) protein levels: hormonal and cytokine control of gene expression in vitro. Hepatology. 1995; 21(6):1675-81. View

20.

Hiam-Galvez K, Allen B, Spitzer M . Systemic immunity in cancer. Nat Rev Cancer. 2021; 21(6):345-359. PMC: 8034277. DOI: 10.1038/s41568-021-00347-z. View