Luminol‐dependent chemiluminescence of human phagocyte cell lines: comparison between DMSO differentiated PLB 985 and HL 60 cells

Abstract

The human promyelocytic leukemia HL 60 and PLB 985 cell lines can differentiate into terminally mature neutrophil-like cells via dimethyl sulfoxide (DMSO) induction. In this study the luminol-dependent chemiluminescence (LCL) of both neutrophil-like cells was analayzed and compared in response to phorbol myristate acetate (PMA) and opsonized zymosan (OZ) stimulants. It was shown that, like human blood neutrophils, both neutrophil-like cells expressed high levels of CD11b, but unlike human blood neutrophils these cells almost lack LCL-detectable intracellular oxidase activity. By studying the pattern of activation to OZ and PMA and priming with GM-CSF, we concluded that there is no difference between the percentage of differentiation and function of DMSO-induced HL 60 and PLB 985. However, the LCL capacity (area under the curve) of DMSO induced PLB 985 cells was higher than that of HL 60 cells in response to both PMA and OZ, which implies a higher capacity to generate reactive oxygen species in PLB 985 cells.

A map of gene expression in neutrophil-like cell lines

“…However, further characterization determined that PLB-985 is actually a sub-line of HL-60 [ 12 ]. PLB-985 cells have similar, but slightly different, properties from those of HL-60 cells, and they are used to study processes including neutrophil ROS production [ 13 ] and chemotaxis [ 14 – 16 ]. These cell lines serve as primary model systems, but even so, they do not fully recapitulate all neutrophil behaviors [ 5 , 17 ].…”

Chemiluminescence of non‐differentiated THP‐1 promonocytes: developing an assay for screening anti‐inflammatory milk proteins and peptides

“…However, in most studies on peripheral blood neutrophils, change of the CL kinetics has not been observed or paid attention to. In human promyelocytic leukemia cell lines HL‐60 and PLB 985 priming with GM‐CSF resulted in an acceleration of OZ‐induced CL reaction, seen as a 25–40% higher CL slope (28). Shorter OZ‐induced CL peaktimes were observed in whole blood neutrophils of diabetic peritoneal dialysis patients with and without acute peritonitis (40).…”

“…Upon differentiation, the THP‐1 cells express increased NADPH‐oxidase activity, cytochrome b content and cell membrane CD11b receptors and acquire manifold capacity to produce superoxide radical during RB (24–27). Indeed, the appearance of RB has been used as an indicator of maturation of THP‐1 cells and leukemic cell lines PLB 985 and HL 60 (28). The transition of normal state circulating monocytes to primed adherent cytokine producing inflammatory cells is a step of critical importance in the development of pathological processes leading to widespread diseases, thus the ability to quantitatively measure RB of nd‐THP‐1 cells is of utmost importance.…”

Multifunctional Micromachines Constructed by Combining Multiple Protein-Based Components with Different Functions

“…The neutrophil-like cells (3 × 10 3 cells), suspended in Hank’s Balanced Salt Solution (HBSS; Invitrogen, CA, USA), were seeded into the multiwell plate (1.9 cm 2 /well), and the diapocynin-loaded turtle machine, prepared by the procedure described in Section , was added into the well, followed by incubation at 37 °C for 1 h. Then, to measure the amounts of the superoxide anion secreted from the cells using chemiluminescence, the cells were collected from the well and suspended in the assay buffer containing the Diogenes, a chemiluminescence probe specific for the superoxide anion, according to the manufacturer’s instructions (Diogenes enhanced superoxide detection kit; National Diagnostics, GA, USA). Secretion of the superoxide anion from neutrophil-like cells was induced by adding the stimulant, phorbol myristate acetate (Wako) at a final concentration of 200 ng mL –1 , , and chemiluminescence was monitored for 10 min using a luminescence reader (AB-2270 Luminescencer Octa; ATTO Co., Ltd., Tokyo, Japan). Chemiluminescence intensity was determined by calculating the area under the chemiluminescence intensity curve (integral chemiluminescence).…”