Heterogeneity of peripheral blood reticulocytes: A flow cytometric analysis with monoclonal antibody HAE9 and thiazole orange

Mechetner, Eugene; Sedmak, Daniel D.; Barth, Rolf F.

doi:10.1002/ajh.2830380110

Cited by 5 publications

(2 citation statements)

References 6 publications

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“…This will contribute to the establishment of a continuous P. vivax culture, since the latter requires a continuous supply of reticulocytes in a large amount. Until now, availability of adequate quantities of reticulocytes was a bottleneck, as they represent only 1% of all circulating cells in the blood stream [24]. Several approaches have been tested to tackle this problem.…”

Section: Discussionmentioning

confidence: 99%

Cryopreserved Reticulocytes Derived from Hematopoietic Stem Cells Can Be Invaded by Cryopreserved Plasmodium vivax Isolates

et al. 2012

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The development of a system for the continuous culture of Plasmodium vivax in vitro would benefit from the use of reticulocytes derived from differentiated hematopoietic stem cells (HCS). At present, the need to use both fresh reticulocytes and fresh P. vivax isolates represents a major obstacle towards this goal, particularly for laboratories located in non-endemic countries. Here, we describe a new method for the cryopreservation of HSC-derived reticulocytes to be used for both P. falciparum and P. vivax invasion tests. Cryopreserved P. falciparum and P. vivax isolates could invade both fresh and cryopreserved HSC-derived reticulocytes with similar efficiency. This new technique allows the storage of HSC-derived reticulocytes which can be used for later invasion tests and represents an important step towards the establishment of a continuous P. vivax culture.

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Section: Discussionmentioning

confidence: 99%

Cryopreserved Reticulocytes Derived from Hematopoietic Stem Cells Can Be Invaded by Cryopreserved Plasmodium vivax Isolates

et al. 2012

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“…Nagakura S, Nakakuma H, Horikawa K, Hidaka M, Kagimoto T, Kawakita M, Tomita M, Takatsuki K: Expression of decay-accelerating factor and CD59 in lymphocyte subsets of healthy individuals and paroxysmal nocturnal hemoglobinuria patients. Am J Hematol 43: 14,1993 9. Fujioka S, Yamada T: Decay-accelerating factor and CD59 expression in peripheral blood cells in aplastic anaemia and report of a case of paroxysmal nocturnal haemoglobinuria secondary to aplastic anaemia.…”

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confidence: 99%

Markedly high population of affected reticulocytes negative for decay- accelerating factor and CD59 in paroxysmal nocturnal hemoglobinuria

Iwamoto

Kawaguchi

Nagakura

et al. 1995

Blood

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Paroxysmal nocturnal hemoglobinuria (PNH) blood cells lack glycosylphosphatidylinositol-anchored membrane proteins such as decay-accelerating factor (DAF) and CD59. This lack is of diagnostic value in PNH. Because reticulocytes in PNH are not yet well characterized, we analyzed reticulocytes obtained from 12 patients with PNH and from 5 healthy volunteers by two-color flow cytometry with a membrane-permeable fluorescent dye, thiazole orange, to identify reticulocytes and monoclonal antibodies to DAF and CD59. Healthy individuals had no affected cells. In all patients, the population of affected reticulocytes negative for DAF and CD59 was markedly higher than the population of affected erythrocytes. Moreover, the population of affected erythrocytes became obviously low in patients who received transfusions and suffered from hemolytic precipitation, whereas the population of affected reticulocytes was unchanged. The persistently high population of affected reticulocytes, despite cytolytic exclusion and an inherently short lifetime, might possibly be explained by relative reticulocytosis caused by an anemia-induced feedback stimulation of erythropoiesis in PNH. Thus, affected reticulocytes could be a reliable marker for the diagnosis of PNH and for the evaluation of erythropoiesis by PNH stem cell.

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Reticulocyte quantification by flow cytometry, image analysis, and manual counting

et al. 1992

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Reticulocyte counting by flow cytometry with thiazole orange was compared to manual or automated counting of new methylene blue stained blood smears. Forty-nine samples were compared for manual counting from randomly chosen clinical samples. Two hundred and eighty-nine samples from bone marrow transplant patients were compared during the period before and through chemo-irradiation and engraftment. The slopes of correlation plots were less than 1 when flow cytometric data were the dependent variable, suggesting that thiazole orange is less sensitive than new methylene blue. In a third study, 407 samples from bone marrow transplant patients were compared after increasing the thiazole orange concentration. The reticulocyte fluorescence distribution was divided into four groups of the brightest (youngest) 40, 60, 80, and 100% of reticulocytes. The slopes from regression analysis were 0.25, 0.49, 0.78, and 1.14, respectively. This demonstrates that thiazole orange is more sensitive than new methylene blue because the window of analysis includes an increased fraction of mature reticulocytes. In addition, the precision of each assay was measured. The rank order of precision from high to low was flow cytometry > image analysis > > manual counting. 0 1992 Wiley-Liss, Inc.

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Heterogeneity of peripheral blood reticulocytes: A flow cytometric analysis with monoclonal antibody HAE9 and thiazole orange

Cited by 5 publications

References 6 publications

Cryopreserved Reticulocytes Derived from Hematopoietic Stem Cells Can Be Invaded by Cryopreserved Plasmodium vivax Isolates

Cryopreserved Reticulocytes Derived from Hematopoietic Stem Cells Can Be Invaded by Cryopreserved Plasmodium vivax Isolates

Markedly high population of affected reticulocytes negative for decay- accelerating factor and CD59 in paroxysmal nocturnal hemoglobinuria

Reticulocyte quantification by flow cytometry, image analysis, and manual counting

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