Detection of Canine Erythrocytes Infected with Babesia gibsoni by Flow Cytometry

Fukata, Tsuneo; Ohnishi, Takafumi; Okuda, Sinnya; Sasai, Kazumi; Baba, Eiichiroh; Arakawa, Akira

doi:10.2307/3283793

Cited by 12 publications

(7 citation statements)

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“…This assay relies on the uptake and metabolic conversion of hydroethidine into ethidium by live parasites. Because conversion does not occur in reticulocytes (2), this assay fails to detect those Babesia species that prefer reticulocytes, such as B. gibsoni (7). In the present study, we adapted the flow cytometric assay that Barkan et al (1) developed to assess parasitemia in mouse models of malaria.…”

Section: Discussionmentioning

confidence: 99%

“…Although considered the gold standard of detection, this test is not ideally suited to quantitatively distinguish reticulocytes from erythrocytes. Inspired by major advances in the diagnosis of malaria using fluorescent nucleic acid stains, flow cytometric assays were developed to assess the viability and growth of B. bovis in red blood cells in vitro (32) and to quantify the percentage of red blood cells infected with B. canis or B. gibsoni in naturally (2) or experimentally infected dogs (7). Although flow cytometry is amenable to multiple and simultaneous detection of surface and intracellular molecules, these studies did not attempt to distinguish reticulocytes from erythrocytes.…”

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confidence: 99%

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Babesia microti Primarily Invades Mature Erythrocytes in Mice

et al. 2006

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Babesia microti is a tick-borne red blood cell parasite that causes babesiosis in people. Its most common vertebrate reservoir is the white-footed mouse. To determine whether B. microti invades reticulocytes, as does the canine pathogen B. gibsoni, we infected the susceptible inbred mouse strains C.B-17.scid and DBA/2 with a clinical isolate of B. microti. Staining of fixed permeabilized red blood cells with 4,6-diamidino-2-phenylindole or YOYO-1, a sensitive nucleic acid stain, revealed parasite nuclei as large bright dots. Flow cytometric analysis indicated that parasite DNA is primarily found in mature erythrocytes that expressed Babesia antigens but not the transferrin receptor CD71. In contrast, CD71-positive reticulocytes rarely contained Babesia nuclei and failed to express Babesia antigens. Accordingly, the frequency of YOYO-1-positive, CD71-negative cells strongly correlated with parasitemia, defined as the frequency of infected red blood cells assessed on Giemsastained blood smears. Importantly, the absolute numbers generated by the two techniques were similar. Parasitemia was modest and transient in DBA/2 mice but intense and sustained in C.B-17.scid mice. In both strains, parasitemia preceded reticulocytosis, but reticulocytes remained refractory to B. microti. In immunocompetent C.B-17 mice, reticulocytosis developed early, despite a marginal and short-lived parasitemia. Likewise, an early reticulocytosis developed in resistant BALB/cBy and B10.D2 mice. These studies establish that B. microti has a tropism for mature erythrocytes. Although reticulocytes are rarely infected, the delayed reticulocytosis in susceptible strains may result from parasite or host activities to limit renewal of the mature erythrocyte pool, thereby preventing an overwhelming parasitemia.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Babesia microti Primarily Invades Mature Erythrocytes in Mice

et al. 2006

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“…This assay, which makes use of the nucleic-acid staining dye YOYO-1 (Molecular Probes), is highly reproducible and correlates very well with counts of B. gibsoni-infected RBCs obtained by microscopy [13]. We have made some minor modifications in this technique.…”

Section: Methodsmentioning

confidence: 99%

Age‐Associated Decline in Resistance toBabesia microtiIs Genetically Determined

Vannier

Borggraefe

Telford³

et al. 2004

J INFECT DIS

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“…The sensitivity of flow cytometry for documentation of acute babesial infections parallels that of conventional light microscopy, with a correlation coefficient as high as 0.97 when used in combination with fluorescent nucleic acid staining 107,109,110 . Flow cytometry has also successfully documented chronic infection, with a higher sensitivity than conventional light microscopy, but lower sensitivity than indirect fluorescent antibody (IFA) testing 111 …”

Section: Diagnosismentioning

confidence: 98%