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Cultivation of Anaplasma ovis in the HL-60 human promyelocytic leukemia cell line
The tick-borne bacterium Anaplasma ovis is a widely distributed pathogen affecting sheep, goats and wild ruminants. Here, the HL-60 human promyelocytic leukemia cell line was used to isolate A. ovis from PCR-positive sheep and goats in Heilongjiang Province, China. Two weeks after inoculation, morul...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5625320/ https://www.ncbi.nlm.nih.gov/pubmed/28928415 http://dx.doi.org/10.1038/emi.2017.70 |
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author | Wei, Ran Liu, Hong-Bo Jongejan, Frans Jiang, Bao-Gui Chang, Qiao-Cheng Fu, Xue Jiang, Jia-Fu Jia, Na Cao, Wu-Chun |
author_facet | Wei, Ran Liu, Hong-Bo Jongejan, Frans Jiang, Bao-Gui Chang, Qiao-Cheng Fu, Xue Jiang, Jia-Fu Jia, Na Cao, Wu-Chun |
author_sort | Wei, Ran |
collection | PubMed |
description | The tick-borne bacterium Anaplasma ovis is a widely distributed pathogen affecting sheep, goats and wild ruminants. Here, the HL-60 human promyelocytic leukemia cell line was used to isolate A. ovis from PCR-positive sheep and goats in Heilongjiang Province, China. Two weeks after inoculation, morulae were observed in cytoplasmic vacuoles in four different HL-60 cultures. Confocal microscopy using a Cy3-labeled A. ovis-specific probe confirmed that the HL-60 cells were infected with A. ovis. Cells from the 6th HL-60 subculture displayed positive fluorescence when incubated with A. ovis antiserum in the indirect fluorescent antibody assay. PCR amplification and sequencing of 16S rRNA, groEL, gltA, msp2 and msp4 Anaplasma genes revealed that the four A. ovis culture isolates were identical. Phylogenetic analysis showed that the sequences clustered with other A. ovis strains but could clearly be distinguished from other Anaplasma species. When the 18th subculture of infected HL-60 cells was examined by electron microscopy, lysosomes were often observed near the vacuoles. After the 24th subculture, Giemsa staining and PCR indicated that the HL-60 cells were negative for A. ovis. Although A. ovis can infect HL-60 cells for only four months, the ability of the organism to infect and multiply in HL-60 cells provides a tool to study intra-erythrocytic Anaplasma and host cell interactions. |
format | Online Article Text |
id | pubmed-5625320 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-56253202017-10-04 Cultivation of Anaplasma ovis in the HL-60 human promyelocytic leukemia cell line Wei, Ran Liu, Hong-Bo Jongejan, Frans Jiang, Bao-Gui Chang, Qiao-Cheng Fu, Xue Jiang, Jia-Fu Jia, Na Cao, Wu-Chun Emerg Microbes Infect Original Article The tick-borne bacterium Anaplasma ovis is a widely distributed pathogen affecting sheep, goats and wild ruminants. Here, the HL-60 human promyelocytic leukemia cell line was used to isolate A. ovis from PCR-positive sheep and goats in Heilongjiang Province, China. Two weeks after inoculation, morulae were observed in cytoplasmic vacuoles in four different HL-60 cultures. Confocal microscopy using a Cy3-labeled A. ovis-specific probe confirmed that the HL-60 cells were infected with A. ovis. Cells from the 6th HL-60 subculture displayed positive fluorescence when incubated with A. ovis antiserum in the indirect fluorescent antibody assay. PCR amplification and sequencing of 16S rRNA, groEL, gltA, msp2 and msp4 Anaplasma genes revealed that the four A. ovis culture isolates were identical. Phylogenetic analysis showed that the sequences clustered with other A. ovis strains but could clearly be distinguished from other Anaplasma species. When the 18th subculture of infected HL-60 cells was examined by electron microscopy, lysosomes were often observed near the vacuoles. After the 24th subculture, Giemsa staining and PCR indicated that the HL-60 cells were negative for A. ovis. Although A. ovis can infect HL-60 cells for only four months, the ability of the organism to infect and multiply in HL-60 cells provides a tool to study intra-erythrocytic Anaplasma and host cell interactions. Nature Publishing Group 2017-09 2017-09-20 /pmc/articles/PMC5625320/ /pubmed/28928415 http://dx.doi.org/10.1038/emi.2017.70 Text en Copyright © 2017 The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Original Article Wei, Ran Liu, Hong-Bo Jongejan, Frans Jiang, Bao-Gui Chang, Qiao-Cheng Fu, Xue Jiang, Jia-Fu Jia, Na Cao, Wu-Chun Cultivation of Anaplasma ovis in the HL-60 human promyelocytic leukemia cell line |
title | Cultivation of Anaplasma ovis in the HL-60 human promyelocytic leukemia cell line |
title_full | Cultivation of Anaplasma ovis in the HL-60 human promyelocytic leukemia cell line |
title_fullStr | Cultivation of Anaplasma ovis in the HL-60 human promyelocytic leukemia cell line |
title_full_unstemmed | Cultivation of Anaplasma ovis in the HL-60 human promyelocytic leukemia cell line |
title_short | Cultivation of Anaplasma ovis in the HL-60 human promyelocytic leukemia cell line |
title_sort | cultivation of anaplasma ovis in the hl-60 human promyelocytic leukemia cell line |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5625320/ https://www.ncbi.nlm.nih.gov/pubmed/28928415 http://dx.doi.org/10.1038/emi.2017.70 |
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