An evaluation of the activity of histidine-rich glycoprotein on differentiated neutrophil-like cells from human cell lines

Published on Dec 27, 2020in Journal of Pharmacology and Experimental Therapeutics3.561
· DOI :10.1124/JPET.120.000182
Yukinori Yoshii3
Estimated H-index: 3
(Okayama University),
Hidenori Wake17
Estimated H-index: 17
(Okayama University)
+ 3 AuthorsMasahiro Nishibori51
Estimated H-index: 51
(Okayama University)
Background: Histidine-rich glycoprotein (HRG) treatment ameliorated the survival rate of septic mice by suppressing excess immunothrombus formation. Although such findings suggested that HRG may be one of the most useful drugs for sepsis, obtaining a stable experimental system to standardize the HRG drug product is difficult to achieve using neutrophils isolated from volunteers. This is due to the short survival time and individual differences of human neutrophils. In the present study, we determined whether the differentiated neutrophil-like cell lines exhibited similar responses to HRG compared to human purified neutrophils. Method: All-trans retinoic acid (ATRA) was employed to induce the differentiation of the human myeloid leukemia cell lines, HL-60 and NB-4. Thereafter, the cells were treated with Hank9s balanced salt solution, human serum albumin, or HRG. The effects of HRG on these cells were evaluated according to cell shape, microcapillary passage, ROS production, neutrophil extracellular traps (NETs) formation, the expression of activated CD11b, and cell viability. Result: HRG maintained the round shape of differentiated neutrophil-like cells, decreased the time required by cells to pass through the microcapillaries, and inhibited ROS production, NETs formation, and the expression of activated CD11b on the cell surface. Moreover, the cells could survive longer in the presence of HRG than the control. Conclusion: The ATRA-induced differentiated cell lines could be used as alternatives to neutrophils to investigate the effects of HRG on neutrophils. This method can thus be used as an essential standardization test in pharmaceutical development. Significance Statement Human neutrophils exhibit varying responses to histidine-rich glycoprotein (HRG); however, all-trans retinoic acid-induced differentiated neutrophil-like cell (NLC) lines can be used as reliably proxies to investigate the effects of HRG on neutrophils. Additionally, these cell lines can be employed in the development of therapies for the treatment of sepsis.
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