Flow cytometric reticulocyte analysis using thiazole orange; clinical experience and technical limitations

Chin‐Yee, Ian; Keeney, Michael; Lohmann, Reinhard

doi:10.1111/j.1365-2257.1991.tb00267.x

Cited by 32 publications

(10 citation statements)

References 9 publications

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“…Despite being enucleated, circulating erythrocytes are phenotypically diverse and range from young erythrocytes (which consist of early and late reticulocytes) to fully matured biconcave erythrocytes (normocytes). Early reticulocytes egress out of hematopoietic organs (primarily the bone marrow) into the peripheral circulation, where they only consist around 1-3% of total human erythrocytes ( Chin-Yee et al., 1991 ). Human reticulocytes undergo extensive cellular changes as they complete their maturation into normocytes ( Malleret et al., 2013 ), such as the loss of ribosomal RNA and the downregulation of many surface proteins ( Wilson et al., 2016 ; Chu et al., 2018 ; Gautier et al., 2018 ).…”

Section: Introductionmentioning

confidence: 99%

Rodent Malaria Erythrocyte Preference Assessment by an Ex Vivo Tropism Assay

Leong

Lee

Rénia

et al. 2021

Front. Cell. Infect. Microbiol.

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Circulating red blood cells consist of young erythrocytes (early and late reticulocytes) and mature erythrocytes (normocytes). The human malaria parasites, Plasmodium falciparum and P. vivax, have a preference to invade reticulocytes during blood-stage infection. Rodent malaria parasites that also prefer reticulocytes could be useful tools to study human malaria reticulocyte invasion. However, previous tropism studies of rodent malaria are inconsistent from one another, making it difficult to compare cell preference of different parasite species and strains. In vivo measurements of cell tropism are also subjected to many confounding factors. Here we developed an ex vivo tropism assay for rodent malaria with highly purified fractions of murine reticulocytes and normocytes. We measured invasion into the different erythrocyte populations using flow cytometry and evaluated the tropism index of the parasite strains. We found that P. berghei ANKA displayed the strongest reticulocyte preference, followed by P. yoelii 17X1.1, whereas P. chabaudi AS and P. vinckei S67 showed mixed tropism. These preferences are intrinsic and were maintained at different reticulocyte and normocyte availabilities. Our study shed light on the true erythrocyte preference of the parasites and paves the way for future investigations on the receptor-ligand interactions mediating erythrocyte tropism.

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Section: Introductionmentioning

confidence: 99%

Rodent Malaria Erythrocyte Preference Assessment by an Ex Vivo Tropism Assay

Leong

Lee

Rénia

et al. 2021

Front. Cell. Infect. Microbiol.

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“…A variety of flow cytometry assays have been developed to achieve this. Earlier assays relied on nucleic acid stains to detect reticulocytes (Chin‐Yee, Keeney, & Lohmann, ; Ware, Rosse, & Hall, ), whereas more recently, antibodies targeting the transferrin receptor (CD71) have been employed (Naumova, Plekhanova, Lugovskaia, Pochtar, & Bugrov, ; Sutherland et al, ). The latter detects immature RBCs (nucleated RBCs and reticulocytes) (Thomson‐Luque et al, ) and does not require the use of nucleic acid stains.…”

Section: Introductionmentioning

confidence: 99%

CD71 improves delineation of PNH type III, PNH type II, and normal immature RBCS in patients with paroxysmal nocturnal hemoglobinuria

Sutherland

Richards

Ortíz

et al. 2019

Cytometry Part B Clinical

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Background The diagnosis of paroxysmal nocturnal hemoglobinuria (PNH) relies on flow cytometric demonstration of loss of glycosyl‐phosphatidyl inositol (GPI)‐anchored proteins from red blood cells (RBC) and white blood cells (WBC). High‐sensitivity multiparameter assays have been developed to detect loss of GPI‐linked structures on PNH neutrophils and monocytes. High‐sensitivity assays to detect PNH phenotypes in RBCs have also been developed that rely on the loss of GPI‐linked CD59 on CD235a‐gated mature RBCs. The latter is used to delineate PNH Type III (total loss of CD59) and PNH Type II RBCs (partial loss of CD59) from normal (Type I) RBCs. However, it is often very difficult to delineate these subsets, especially in patients with large PNH clones who continue to receive RBC transfusions, even while on eculizumab therapy. Methods We have added allophycocyanin (APC)‐conjugated CD71 to the existing CD235aFITC/CD59PE RBC assay allowing simultaneous delineation and quantification of PNH Type III and Type II immature RBCs (iRBCs). Results We analyzed 24 medium to large‐clone PNH samples (>10% PNH WBC clone size) for PNH Neutrophil, PNH Monocyte, Type III and Type II PNH iRBCs, and where possible, Type III and Type II PNH RBCs. The ability to delineate PNH Type III, Type II, and Type I iRBCs was more objective compared to that in mature RBCs. Additionally, total PNH iRBC clone sizes were very similar to PNH WBC clone sizes. Conclusions Addition of CD71 significantly improves the ability to analyze PNH clone sizes in the RBC lineage, regardless of patient hemolytic and/or transfusion status.

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“…Nothing has been published about potential changes in blood count or differential blood count because of changed life styles over the course of the last decades, which calls into question the transferability of old reference ranges to today's population. However, results from external quality assessment schemes show clear analyzer-specific differences do exist [24][25][26][27][28]. Results from hospital patient groups (uncertain health status) [21,22] or preselected blood donor groups must also be looked at with a critical eye.…”

Section: Introductionmentioning

confidence: 99%

Multi-centric determination of reference ranges for automated blood counts¹

Nebe

Bentzien

Bruegel

et al. 2011

LaboratoriumsMedizin

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The analysis of blood cell count including differential, is the most frequent laboratory test and has still essential diagnostic value in the diagnosis of primary or secondary diseases of the hematopoietic system. The interpretation is based on a comparison with age matched reference intervals, despite the fact that a comparison with the patients own preceding values allows a more precise statement. In the present study, blood counts and differential were performed in 1158 healthy males and females with an age ranging from 16 to 75 years were investigated using current hematology systems. The donors were examined by personnel physicians and those examining blood donors in transfusion medicine and in addition were asked via standardized questionnaire. The study resulted in reference ranges for the complete blood count, the differential blood count and reticulocytes. For several analytes, age-, gender-or analyzerspecific reference ranges were obtained. We found an age dependent decrease of the erythrocyte concentration and an increase for MCH and MCV as well as for basophils. These findings could be important for the diagnosis of diseases of the elderly like like myeloproliferative diseases (MPN) and myelodysplasias (MDS). Gender specific changes could be shown for the hemoglobulin-, the erythrocyte-and reticulocyte concentration, as well as for hematocrit and MCHC, being reduced in females. On the other hand, females showed significant higher levels for platelets and leukocytes, based on an increase of neutrophils and lymphocytes. The leukocytosis of smokers was confirmed for neutrophils and lymphocytes but lower than one could expect from recent studies. A multicentric study shows a broader distribution compared to unicentric data but this probably better reflects the reality and improves the applicability of the reference ranges.

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Flow cytometric reticulocyte analysis using thiazole orange; clinical experience and technical limitations

Cited by 32 publications

References 9 publications

Rodent Malaria Erythrocyte Preference Assessment by an Ex Vivo Tropism Assay

Rodent Malaria Erythrocyte Preference Assessment by an Ex Vivo Tropism Assay

CD71 improves delineation of PNH type III, PNH type II, and normal immature RBCS in patients with paroxysmal nocturnal hemoglobinuria

Multi-centric determination of reference ranges for automated blood counts¹

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Flow cytometric reticulocyte analysis using thiazole orange; clinical experience and technical limitations

Cited by 32 publications

References 9 publications

Rodent Malaria Erythrocyte Preference Assessment by an Ex Vivo Tropism Assay

Rodent Malaria Erythrocyte Preference Assessment by an Ex Vivo Tropism Assay

CD71 improves delineation of PNH type III, PNH type II, and normal immature RBCS in patients with paroxysmal nocturnal hemoglobinuria

Multi-centric determination of reference ranges for automated blood counts1

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Multi-centric determination of reference ranges for automated blood counts¹