A novel flow cytometry‐based method of analyzing Heinz bodies

Palasuwan, Duangdao; Palasuwan, Attakorn; Charoensappakit, Awirut; Noulsri, Egarit

doi:10.1111/ijlh.12576

Cited by 4 publications

(4 citation statements)

References 24 publications

(37 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The following three markers were evaluated by flow cytometry: PS, intracellular ROS and autofluorescence. Increases in PS on the surface of RBCs are an indicator of membrane damage, intracellular ROS provides information on oxidative stress within the RBC and elevation of autofluorescence has been linked to Heinz body formation and clearance 31 . G6PDd‐huRBC engrafted mice received one dose of VC, PQ (15 mpk) or TQ (10 mpk).…”

Section: Resultsmentioning

confidence: 99%

“…Increases in PS on the surface of RBCs are an indicator of membrane damage, intracellular ROS provides information on oxidative stress within the RBC and elevation of autofluorescence has been linked to Heinz body formation and clearance. 31 G6PDd‐huRBC engrafted mice received one dose of VC, PQ (15 mpk) or TQ (10 mpk). RBCs were analysed at 24‐ and 48‐ h post‐dosing.…”

Section: Resultsmentioning

confidence: 99%

“…ROS levels were determined by staining with 2’,7’‐dichlorodihydrofluorescein‐diacetate (DCFDA) (Sigma, Saint Louis, MO) in Ringer solution (Boston BioProducts, Ashland, MA) as previously described 30 . Autofluorescence was measured by the methods previously described 31 …”

Section: Methodsmentioning

confidence: 99%

“…30 Autofluorescence was measured by the methods previously described. 31 All flow cytometry was performed using a Cytek ® Northern Lights flow cytometer, 10 5 events were collected and results were analysed using FlowJo™ Software (BD) with graphs generated using GraphPad Prism 8.0 software (GraphPad Software, Inc., San Diego, CA). Sample gating strategy for Annexin-PI and autofluorescence is provided in Figure S1.…”

Section: Flow Cytometric Analysis Of G6pdd-hurbcmentioning

confidence: 99%

See 3 more Smart Citations

Mechanisms of 8‐aminoquinoline induced haemolytic toxicity in a G6PDd humanized mouse model

Flaherty

Strauch

Maktabi

et al. 2022

J Cellular Molecular Medi

View full text Add to dashboard Cite

Primaquine (PQ) and Tafenoquine (TQ) are clinically important 8‐aminoquinolines (8‐AQ) used for radical cure treatment of P. vivax infection, known to target hepatic hypnozoites. 8‐AQs can trigger haemolytic anaemia in individuals with glucose‐6‐phosphate dehydrogenase deficiency (G6PDd), yet the mechanisms of haemolytic toxicity remain unknown. To address this issue, we used a humanized mouse model known to predict haemolytic toxicity responses in G6PDd human red blood cells (huRBCs). To evaluate the markers of eryptosis, huRBCs were isolated from mice 24–48 h post‐treatment and analysed for effects on phosphatidylserine (PS), intracellular reactive oxygen species (ROS) and autofluorescence. Urinalysis was performed to evaluate the occurrence of intravascular and extravascular haemolysis. Spleen and liver tissue harvested at 24 h and 5–7 days post‐treatment were stained for the presence of CD169+ macrophages, F4/80+ macrophages, Ter119+ mouse RBCs, glycophorin A+ huRBCs and murine reticulocytes (muRetics). G6PDd‐huRBCs from PQ/TQ treated mice showed increased markers for eryptosis as early as 24 h post‐treatment. This coincided with an early rise in levels of muRetics. Urinalysis revealed concurrent intravascular and extravascular haemolysis in response to PQ/TQ. Splenic CD169+ macrophages, present in all groups at day 1 post‐dosing were eliminated by days 5–7 in PQ/TQ treated mice only, while liver F4/80 macrophages and iron deposits increased. Collectively, our data suggest 8‐AQ treated G6PDd‐huRBCs have early physiological responses to treatment, including increased markers for eryptosis indicative of oxidative stress, resulting in extramedullary haematopoiesis and loss of splenic CD169+ macrophages, prompting the liver to act as the primary site of clearance.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%