Extracellular HMGB1 promotes the migration of cord Blood CD34⁺ cells via SDF-1/CXCR-4 axis.
10.7534/j.issn.1009-2137.2014.05.044
- Author:
Lu-Lu YANG
1
;
Zi-Min SUN
1
;
Xin LIU
2
;
Xiao-Yu ZHU
3
;
Xing-Bing WANG
1
;
Jian WANG
1
Author Information
1. Department of Hematology, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei 23001, Auhui Province, China.
2. Department of Hematology, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei 23001, Auhui Province, China. E-mail:lxinahf@sina.com.
3. Department of Hematology, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei 23001, Auhui Province, China. E-mail: bayinhexy@126.com.
- Publication Type:Journal Article
- MeSH:
Antigens, CD34;
metabolism;
Cell Movement;
Chemokine CXCL12;
metabolism;
Fetal Blood;
cytology;
metabolism;
Flow Cytometry;
HMGB1 Protein;
metabolism;
Humans;
Receptors, CXCR4;
metabolism
- From:
Journal of Experimental Hematology
2014;22(5):1415-1421
- CountryChina
- Language:Chinese
-
Abstract:
This study was aimed to investigate the effect of high mobility group box1(HMGB1) and/or stromal cell derived factor-1(SDF-1) on the migration of cord blood CD34⁺ cells, and to explore whether HMGB1 promotes cord blood CD34⁺ cell migration via SDF-1/CXCR4 axis. Cord blood mononuclear cells were isolated by Ficoll-Paque density centrifugation, CD34⁺ cells were collected by a positive immunoselection procedure (CD34 MicroBeads) according to the manufacturer's instructions, the purity of the isolated CD34⁺ cells was detected by flow cytometry. In vitro chemotaxis assays were performed using Transwell cell chambers to detect cells migration. 1 × 10⁵ cells/well cord blood CD34⁺ cells were added into the upper chambers. Different concentrations of HMGB1 and/or SDF-1 (0, 10, 25, 50, 100, 200 ng/ml) were used to detect the optimal concentrations of HMGB1 and/or SDF-1 for inducing migration of cord blood CD34⁺ cells. Freshly isolated cord blood CD34⁺ cells express CXCR4 (SDF-1 receptor), and HMGB1 receptor TLR-2,TLR-4 and RAGE. To explore which receptors were required for the synergy of HGMB1 and/or SDF-1 on cells migration, the anti-SDF-1, anti-CXCR4 and anti-RAGE antibodies were used to detect the effect of HGMB1 alone or with SDF-1 on cord blood CD34⁺ cells migration. The results showed that the purity of CD34⁺ cells isolated from cord blood mononuclear cells by magnetic cell sorting was 97.40 ± 1.26%, the 25 ng/ml SDF-1 did not induce migration of cord blood CD34⁺ cells, whereas the optimal migration was observed at 100 ng/ml. HMGB1 alone did not induce migration up to 100 ng/ml. The dose test found that the the best synergistic concentrations for cells migration were 100 ng/ml HMGB1 combined with 50 ng/ml SDF-1. The blocking test showed that both the anti-SDF-1 (4 µg/ml) and anti-CXCR4 (5 µg/ml) antibodies could block cell migration induced by HMGB1 or combined with SDF-1, but the cord blood CD34⁺ cells in the presence of anti-RAGE, anti-TLR-2 and anti-TLR-4 antibodies did not modify the response to SDF-1 in the presence of HMGB1. It is concluded that both HMGB1 and SDF-1 can induce cord blood CD34⁺ cells migration, HMGB1 enhances SDF-1-induced migration exclusively via CXCR4 and in a RAGE and TLR receptors-independent manner, the exact mechanism needs to be further explored.
