Toxoplasma gondii tachyzoite-infected peripheral blood mononuclear cells are enriched in mouse lungs and liver

[Display omitted] •We developed a method to estimate tachyzoite and PBMC numbers in solid organs.•The PBMC infection rates in several mouse organs were compared.•The tachyzoite-infected PBMCs were enriched in mouse lungs and liver. The intracellular parasite Toxoplasma gondii is thought to dissemina...

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Veröffentlicht in:Experimental parasitology 2013-06, Vol.134 (2), p.160-164
Hauptverfasser: Unno, Akihiro, Kachi, Seira, Batanova, Tatiana A., Ohno, Tamio, Elhawary, Nagwa, Kitoh, Katsuya, Takashima, Yasuhiro
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container_end_page 164
container_issue 2
container_start_page 160
container_title Experimental parasitology
container_volume 134
creator Unno, Akihiro
Kachi, Seira
Batanova, Tatiana A.
Ohno, Tamio
Elhawary, Nagwa
Kitoh, Katsuya
Takashima, Yasuhiro
description [Display omitted] •We developed a method to estimate tachyzoite and PBMC numbers in solid organs.•The PBMC infection rates in several mouse organs were compared.•The tachyzoite-infected PBMCs were enriched in mouse lungs and liver. The intracellular parasite Toxoplasma gondii is thought to disseminate throughout the host by circulation of tachyzoite-infected leukocytes in the blood, and adherence and migration of such leukocytes into solid tissues. However, it is unclear whether T. gondii-infected leukocytes can migrate to solid organs via the general circulation. In this study, we developed a real-time quantitative PCR (qRT-PCR) method to determine the rate of infection of peripheral blood mononuclear cells (PBMCs) flowing into and remaining within solid organs in mice. A transgenic T. gondii parasite line derived from the PLK strain that expresses DsRed Express, and transgenic green fluorescent protein-positive PBMCs, were used for these experiments. Tachyzoite-infected PBMCs were injected into mouse tail veins and qRT-PCR was used to measure the infection rates of the PBMCs remaining in the lungs, liver, spleen and brain. We found that the PBMCs in the lungs and liver had statistically higher infection rates than that of the original inoculum; this difference was statistically significant. However, the PBMC infection rate in the spleen showed no such enhancement. These results show that tachyzoite-infected PBMCs in the general circulation remain in the lungs and liver more effectively than non-infected PBMCs.
doi_str_mv 10.1016/j.exppara.2013.03.006
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The intracellular parasite Toxoplasma gondii is thought to disseminate throughout the host by circulation of tachyzoite-infected leukocytes in the blood, and adherence and migration of such leukocytes into solid tissues. However, it is unclear whether T. gondii-infected leukocytes can migrate to solid organs via the general circulation. In this study, we developed a real-time quantitative PCR (qRT-PCR) method to determine the rate of infection of peripheral blood mononuclear cells (PBMCs) flowing into and remaining within solid organs in mice. A transgenic T. gondii parasite line derived from the PLK strain that expresses DsRed Express, and transgenic green fluorescent protein-positive PBMCs, were used for these experiments. Tachyzoite-infected PBMCs were injected into mouse tail veins and qRT-PCR was used to measure the infection rates of the PBMCs remaining in the lungs, liver, spleen and brain. We found that the PBMCs in the lungs and liver had statistically higher infection rates than that of the original inoculum; this difference was statistically significant. However, the PBMC infection rate in the spleen showed no such enhancement. These results show that tachyzoite-infected PBMCs in the general circulation remain in the lungs and liver more effectively than non-infected PBMCs.</description><identifier>ISSN: 0014-4894</identifier><identifier>EISSN: 1090-2449</identifier><identifier>DOI: 10.1016/j.exppara.2013.03.006</identifier><identifier>PMID: 23538031</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Animals ; brain ; cell movement ; Cercopithecus aethiops ; Circulation ; DNA, Protozoan - isolation &amp; purification ; fluorescence ; genetically modified organisms ; Green Fluorescent Proteins ; Heart Ventricles - parasitology ; Heart Ventricles - pathology ; inoculum ; Leukocytes, Mononuclear - cytology ; Leukocytes, Mononuclear - parasitology ; liver ; Liver - parasitology ; Liver - pathology ; Luminescent Agents ; Lung - parasitology ; Lung - pathology ; lungs ; Male ; Mice ; Mice, Inbred C57BL ; mononuclear leukocytes ; parasites ; parasitology ; Peripheral blood mononuclear cells (PBMCs) ; quantitative polymerase chain reaction ; Real-Time Polymerase Chain Reaction ; Real-time quantitative PCR (qRT-PCR) ; reverse transcriptase polymerase chain reaction ; spleen ; Spleen - parasitology ; Spleen - pathology ; Tachyzoite ; tail ; tissues ; Toxoplasma - genetics ; Toxoplasma - isolation &amp; purification ; Toxoplasma - physiology ; Toxoplasma gondii ; Toxoplasmosis, Animal - blood ; Toxoplasmosis, Animal - parasitology ; Toxoplasmosis, Animal - pathology ; Vero Cells</subject><ispartof>Experimental parasitology, 2013-06, Vol.134 (2), p.160-164</ispartof><rights>2013 Elsevier Inc.</rights><rights>Copyright © 2013 Elsevier Inc. 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The intracellular parasite Toxoplasma gondii is thought to disseminate throughout the host by circulation of tachyzoite-infected leukocytes in the blood, and adherence and migration of such leukocytes into solid tissues. However, it is unclear whether T. gondii-infected leukocytes can migrate to solid organs via the general circulation. In this study, we developed a real-time quantitative PCR (qRT-PCR) method to determine the rate of infection of peripheral blood mononuclear cells (PBMCs) flowing into and remaining within solid organs in mice. A transgenic T. gondii parasite line derived from the PLK strain that expresses DsRed Express, and transgenic green fluorescent protein-positive PBMCs, were used for these experiments. Tachyzoite-infected PBMCs were injected into mouse tail veins and qRT-PCR was used to measure the infection rates of the PBMCs remaining in the lungs, liver, spleen and brain. We found that the PBMCs in the lungs and liver had statistically higher infection rates than that of the original inoculum; this difference was statistically significant. However, the PBMC infection rate in the spleen showed no such enhancement. These results show that tachyzoite-infected PBMCs in the general circulation remain in the lungs and liver more effectively than non-infected PBMCs.</description><subject>Animals</subject><subject>brain</subject><subject>cell movement</subject><subject>Cercopithecus aethiops</subject><subject>Circulation</subject><subject>DNA, Protozoan - isolation &amp; purification</subject><subject>fluorescence</subject><subject>genetically modified organisms</subject><subject>Green Fluorescent Proteins</subject><subject>Heart Ventricles - parasitology</subject><subject>Heart Ventricles - pathology</subject><subject>inoculum</subject><subject>Leukocytes, Mononuclear - cytology</subject><subject>Leukocytes, Mononuclear - parasitology</subject><subject>liver</subject><subject>Liver - parasitology</subject><subject>Liver - pathology</subject><subject>Luminescent Agents</subject><subject>Lung - parasitology</subject><subject>Lung - pathology</subject><subject>lungs</subject><subject>Male</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>mononuclear leukocytes</subject><subject>parasites</subject><subject>parasitology</subject><subject>Peripheral blood mononuclear cells (PBMCs)</subject><subject>quantitative polymerase chain reaction</subject><subject>Real-Time Polymerase Chain Reaction</subject><subject>Real-time quantitative PCR (qRT-PCR)</subject><subject>reverse transcriptase polymerase chain reaction</subject><subject>spleen</subject><subject>Spleen - parasitology</subject><subject>Spleen - pathology</subject><subject>Tachyzoite</subject><subject>tail</subject><subject>tissues</subject><subject>Toxoplasma - genetics</subject><subject>Toxoplasma - isolation &amp; purification</subject><subject>Toxoplasma - physiology</subject><subject>Toxoplasma gondii</subject><subject>Toxoplasmosis, Animal - blood</subject><subject>Toxoplasmosis, Animal - parasitology</subject><subject>Toxoplasmosis, Animal - pathology</subject><subject>Vero Cells</subject><issn>0014-4894</issn><issn>1090-2449</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkcFuEzEQhi0EoqHwCICPXDaM1_ZmfUKoKhSpEgfas-W1J4kjr73Yu1XL0-MogWulkXzwNzO_viHkPYM1A9Z9PqzxcZpMNusWGF9DLehekBUDBU0rhHpJVgBMNKJX4oK8KeUAAD1rxWty0XLJe-BsReJdekxTMGU0dJei857Oxu6f_iQ_Y-PjFu2Mjk6Y_bTHbAIdQkqOjimmuNiAJlOLIRRqMlKM2dt95X2sxFKQhiXu6l90NPgHzG_Jq60JBd-d30ty_-367uqmuf35_cfV19vGCinnpuOuM1IprIktG1rHrbUb0Qtne6YkExINgBICO7Q9R8WsZFb0A2dWsQH4Jfl0mjvl9HvBMuvRl2NOE7Hm0oxLYFJs-rai8oTanErJuNVT9qPJT5qBPqrWB31WrY-qNdSCrvZ9OK9YhhHd_65_bivw8QRsTdJml33R97_qBFnPsuF9d1z95URgVfHgMetiPUaLzufqXbvknwnxF_TPnQc</recordid><startdate>20130601</startdate><enddate>20130601</enddate><creator>Unno, Akihiro</creator><creator>Kachi, Seira</creator><creator>Batanova, Tatiana A.</creator><creator>Ohno, Tamio</creator><creator>Elhawary, Nagwa</creator><creator>Kitoh, Katsuya</creator><creator>Takashima, Yasuhiro</creator><general>Elsevier Inc</general><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20130601</creationdate><title>Toxoplasma gondii tachyzoite-infected peripheral blood mononuclear cells are enriched in mouse lungs and liver</title><author>Unno, Akihiro ; Kachi, Seira ; Batanova, Tatiana A. ; Ohno, Tamio ; Elhawary, Nagwa ; Kitoh, Katsuya ; Takashima, Yasuhiro</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c455t-63d6a599e008c1b2d3ccc7484dc8195145ea00944e6ec83e91c51c48b31c91b03</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Animals</topic><topic>brain</topic><topic>cell movement</topic><topic>Cercopithecus aethiops</topic><topic>Circulation</topic><topic>DNA, Protozoan - isolation &amp; purification</topic><topic>fluorescence</topic><topic>genetically modified organisms</topic><topic>Green Fluorescent Proteins</topic><topic>Heart Ventricles - parasitology</topic><topic>Heart Ventricles - pathology</topic><topic>inoculum</topic><topic>Leukocytes, Mononuclear - cytology</topic><topic>Leukocytes, Mononuclear - parasitology</topic><topic>liver</topic><topic>Liver - parasitology</topic><topic>Liver - pathology</topic><topic>Luminescent Agents</topic><topic>Lung - parasitology</topic><topic>Lung - pathology</topic><topic>lungs</topic><topic>Male</topic><topic>Mice</topic><topic>Mice, Inbred C57BL</topic><topic>mononuclear leukocytes</topic><topic>parasites</topic><topic>parasitology</topic><topic>Peripheral blood mononuclear cells (PBMCs)</topic><topic>quantitative polymerase chain reaction</topic><topic>Real-Time Polymerase Chain Reaction</topic><topic>Real-time quantitative PCR (qRT-PCR)</topic><topic>reverse transcriptase polymerase chain reaction</topic><topic>spleen</topic><topic>Spleen - parasitology</topic><topic>Spleen - pathology</topic><topic>Tachyzoite</topic><topic>tail</topic><topic>tissues</topic><topic>Toxoplasma - genetics</topic><topic>Toxoplasma - isolation &amp; purification</topic><topic>Toxoplasma - physiology</topic><topic>Toxoplasma gondii</topic><topic>Toxoplasmosis, Animal - blood</topic><topic>Toxoplasmosis, Animal - parasitology</topic><topic>Toxoplasmosis, Animal - pathology</topic><topic>Vero Cells</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Unno, Akihiro</creatorcontrib><creatorcontrib>Kachi, Seira</creatorcontrib><creatorcontrib>Batanova, Tatiana A.</creatorcontrib><creatorcontrib>Ohno, Tamio</creatorcontrib><creatorcontrib>Elhawary, Nagwa</creatorcontrib><creatorcontrib>Kitoh, Katsuya</creatorcontrib><creatorcontrib>Takashima, Yasuhiro</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Experimental parasitology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Unno, Akihiro</au><au>Kachi, Seira</au><au>Batanova, Tatiana A.</au><au>Ohno, Tamio</au><au>Elhawary, Nagwa</au><au>Kitoh, Katsuya</au><au>Takashima, Yasuhiro</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Toxoplasma gondii tachyzoite-infected peripheral blood mononuclear cells are enriched in mouse lungs and liver</atitle><jtitle>Experimental parasitology</jtitle><addtitle>Exp Parasitol</addtitle><date>2013-06-01</date><risdate>2013</risdate><volume>134</volume><issue>2</issue><spage>160</spage><epage>164</epage><pages>160-164</pages><issn>0014-4894</issn><eissn>1090-2449</eissn><abstract>[Display omitted] •We developed a method to estimate tachyzoite and PBMC numbers in solid organs.•The PBMC infection rates in several mouse organs were compared.•The tachyzoite-infected PBMCs were enriched in mouse lungs and liver. The intracellular parasite Toxoplasma gondii is thought to disseminate throughout the host by circulation of tachyzoite-infected leukocytes in the blood, and adherence and migration of such leukocytes into solid tissues. However, it is unclear whether T. gondii-infected leukocytes can migrate to solid organs via the general circulation. In this study, we developed a real-time quantitative PCR (qRT-PCR) method to determine the rate of infection of peripheral blood mononuclear cells (PBMCs) flowing into and remaining within solid organs in mice. A transgenic T. gondii parasite line derived from the PLK strain that expresses DsRed Express, and transgenic green fluorescent protein-positive PBMCs, were used for these experiments. Tachyzoite-infected PBMCs were injected into mouse tail veins and qRT-PCR was used to measure the infection rates of the PBMCs remaining in the lungs, liver, spleen and brain. We found that the PBMCs in the lungs and liver had statistically higher infection rates than that of the original inoculum; this difference was statistically significant. However, the PBMC infection rate in the spleen showed no such enhancement. These results show that tachyzoite-infected PBMCs in the general circulation remain in the lungs and liver more effectively than non-infected PBMCs.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>23538031</pmid><doi>10.1016/j.exppara.2013.03.006</doi><tpages>5</tpages></addata></record>
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source MEDLINE; Access via ScienceDirect (Elsevier)
subjects Animals
brain
cell movement
Cercopithecus aethiops
Circulation
DNA, Protozoan - isolation & purification
fluorescence
genetically modified organisms
Green Fluorescent Proteins
Heart Ventricles - parasitology
Heart Ventricles - pathology
inoculum
Leukocytes, Mononuclear - cytology
Leukocytes, Mononuclear - parasitology
liver
Liver - parasitology
Liver - pathology
Luminescent Agents
Lung - parasitology
Lung - pathology
lungs
Male
Mice
Mice, Inbred C57BL
mononuclear leukocytes
parasites
parasitology
Peripheral blood mononuclear cells (PBMCs)
quantitative polymerase chain reaction
Real-Time Polymerase Chain Reaction
Real-time quantitative PCR (qRT-PCR)
reverse transcriptase polymerase chain reaction
spleen
Spleen - parasitology
Spleen - pathology
Tachyzoite
tail
tissues
Toxoplasma - genetics
Toxoplasma - isolation & purification
Toxoplasma - physiology
Toxoplasma gondii
Toxoplasmosis, Animal - blood
Toxoplasmosis, Animal - parasitology
Toxoplasmosis, Animal - pathology
Vero Cells
title Toxoplasma gondii tachyzoite-infected peripheral blood mononuclear cells are enriched in mouse lungs and liver
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