Name: in vivo imaging of transgenic leishmania parasites in a live host
Uploaded: 2010-07-27T17:00:00
Description: Watch this Scientific Journal Video about In vivo Imaging of Transgenic Leishmania Parasites in a Live Host at JoVE.com

Detta arbete har finansierats delvis av en Merit Review bidrag från Department of Veterans Affairs, genom NIH anslag AI045540, AI067874, AI076233-01 och AI080801 (MEW), och genom AI29646 (SMB). Arbetet utfördes delvis under finansiering av CT och JG genom NIH T32 AI07511.

1. Infektion av små djur med transgena Leishmania 1. Parasit linjer Transgena Leishmania spp.. parasiter uttrycka luciferas genereras med en episomal eller en integrerande vektor som rapporterats. 1 2 klonal linjer är att föredra. Två viktiga punkter: (A)-luciferas är att föredra framför episomal luciferas, eftersom det i teorin dessa parasiter linjer bör hålla oss till den transgenen i frånvaro av dr...

<ol>
	<li>Capul, A. A., Barron, T., Dobson, D. E., Turco, S. J., Beverley, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two+functionally+divergent+UDP-Gal+nucleotide+sugar+transporters+participate+in+phosphoglycan+synthesis+in+Leishmania+major.">Two functionally divergent UDP-Gal nucleotide sugar transporters participate in phosphoglycan synthesis in Leishmania major.</a> J Biol Chem. 282, 14006-14017 (2007).</li><li>LeBowitz, J. H., Coburn, C. M., McMahon-Pratt, D., Beverley, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+stable+Leishmania+expression+vector+and+application+to+the+study+of+parasite+surface+antigen+genes.">Development of a stable Leishmania expression vector and application to the study of parasite surface antigen genes.</a> Proc Natl Acad Sci U S A. 87, 9736-9740 (1990).</li><li>Mureev, S., Kushnir, S., Kolesnikov, A. A., Breitling, R., Alexandrov, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Construction+and+analysis+of+Leishmania+tarentolae+transgenic+strains+free+of+selection+markers.">Construction and analysis of Leishmania tarentolae transgenic strains free of selection markers.</a> Mol Biochem Parasitol. 155, 71-83 (2007).</li><li>Chakkalath, H. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Priming+of+a+beta-galactosidase+(beta-GAL)-specific+type+1+response+in+BALB/c+mice+infected+with+beta-GAL-transfected+Leishmania+major.">Priming of a beta-galactosidase (beta-GAL)-specific type 1 response in BALB/c mice infected with beta-GAL-transfected Leishmania major.</a> Infect Immun. 68, 809-814 (2000).</li><li>Sacks, D. L., Perkins, P. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+an+infective+stage+of+Leishmania+promastigotes.">Identification of an infective stage of Leishmania promastigotes.</a> Science. 223, 1417-1419 (1984).</li><li>da Silva, R., Sacks, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metacyclogenesis+is+a+major+determinant+of+Leishmania+promastigote+virulence+and+attenuation.">Metacyclogenesis is a major determinant of Leishmania promastigote virulence and attenuation.</a> Infect Immun. 55, 2802-2806 (1987).</li><li>Spath, G. F., Beverley, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+lipophosphoglycan-independent+method+for+isolation+of+infective+Leishmania+metacyclic+promastigotes+by+density+gradient+centrifugation.">A lipophosphoglycan-independent method for isolation of infective Leishmania metacyclic promastigotes by density gradient centrifugation.</a> Exp Parasitol. 99, 97-103 (2001).</li><li>Scott, P., Caspar, P., Sher, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protection+against+Leishmania+major+in+BALB/c+mice+by+adoptive+transfer+of+a+T+cell+clone+recognizing+a+low+molecular+weight+antigen+released+by+promastigotes.">Protection against Leishmania major in BALB/c mice by adoptive transfer of a T cell clone recognizing a low molecular weight antigen released by promastigotes.</a> J Immunol. 144, 1075-1079 (1990).</li><li>Belkaid, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+interleukin+(IL)-10+in+the+persistence+of+Leishmania+major+in+the+skin+after+healing+and+the+therapeutic+potential+of+anti-IL-10+receptor+antibody+for+sterile+cure.">The role of interleukin (IL)-10 in the persistence of Leishmania major in the skin after healing and the therapeutic potential of anti-IL-10 receptor antibody for sterile cure.</a> J Exp Med. 194, 1497-1506 (2001).</li><li>Wilson, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Local+suppression+of+IFN-gamma+in+hepatic+granulomas+correlates+with+tissue-specific+replication+of+Leishmania+chagasi.">Local suppression of IFN-gamma in hepatic granulomas correlates with tissue-specific replication of Leishmania chagasi.</a> J Immunol. 156, 2231-2239 (1996).</li><li>McElrath, M. J., Murray, H. W., Cohn, Z. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+dynamics+of+granuloma+formation+in+experimental+visceral+leishmaniasis.">The dynamics of granuloma formation in experimental visceral leishmaniasis.</a> J Exp Med. 167, 1927-1937 (1988).</li><li>Ato, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Loss+of+dendritic+cell+migration+and+impaired+resistance+to+Leishmania+donovani+infection+in+mice+deficient+in+CCL19+and+CCL21.">Loss of dendritic cell migration and impaired resistance to Leishmania donovani infection in mice deficient in CCL19 and CCL21.</a> J Immunol. 176, 5486-5493 (2006).</li><li>Ahmed, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intradermal+infection+model+for+pathogenesis+and+vaccine+studies+of+murine+visceral+leishmaniasis.">Intradermal infection model for pathogenesis and vaccine studies of murine visceral leishmaniasis.</a> Infect Immun. 71, 401-410 (2003).</li><li>Kamala, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hock+immunization:+a+humane+alternative+to+mouse+footpad+injections.">Hock immunization: a humane alternative to mouse footpad injections.</a> J Immunol Methods. 328, 204-214 (2007).</li><li>Lang, T., Goyard, S., Lebastard, M., Milon, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bioluminescent+Leishmania+expressing+luciferase+for+rapid+and+high+throughput+screening+of+drugs+acting+on+amastigote-harbouring+macrophages+and+for+quantitative+real-time+monitoring+of+parasitism+features+in+living+mice.">Bioluminescent Leishmania expressing luciferase for rapid and high throughput screening of drugs acting on amastigote-harbouring macrophages and for quantitative real-time monitoring of parasitism features in living mice.</a> Cell Microbiol. 7, 383-392 (2005).</li><li>Brittingham, A., Miller, M. A., Donelson, J. E., Wilson, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+GP63+mRNA+stability+in+promastigotes+of+virulent+and+attenuated+Leishmania+chagasi.">Regulation of GP63 mRNA stability in promastigotes of virulent and attenuated Leishmania chagasi.</a> Mol Biochem Parasitol. 112, 51-59 (2001).</li><li>Roy, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Episomal+and+stable+expression+of+the+luciferase+reporter+gene+for+quantifying+Leishmania+spp.+infections+in+macrophages+and+in+animal+models.">Episomal and stable expression of the luciferase reporter gene for quantifying Leishmania spp. infections in macrophages and in animal models.</a> Mol Biochem Parasitol. 110, 195-206 (2000).</li><li>Contag, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photonic+detection+of+bacterial+pathogens+in+living+hosts.">Photonic detection of bacterial pathogens in living hosts.</a> Mol Microbiol. 18, 593-603 (1995).</li><li>Hardy, J., Margolis, J. J., Contag, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induced+Biliary+Excretion+of+Listeria+monocytogenes.">Induced Biliary Excretion of Listeria monocytogenes.</a> Infect. Immun. 74, 1819-1827 (2006).</li><li>Lecoeur, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimization+of+Topical+Therapy+for+Leishmania+major+Localized+Cutaneous+Leishmaniasis+Using+a+Reliable+C57BL/6+Model.">Optimization of Topical Therapy for Leishmania major Localized Cutaneous Leishmaniasis Using a Reliable C57BL/6 Model.</a> PLoS Negl Trop Dis. 1, e34-e34 (2007).</li><li>Lang, T., Lecoeur, H., Prina, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+Leishmania+development+in+their+host+cells.">Imaging Leishmania development in their host cells.</a> Trends Parasitol. 25, 464-473 (2009).</li></ol>

In vivo imaging-system (IVIS) ger en metod för hela djuret avbildning eller in vivo imaging experimentell infektion modeller av olika former av leishmaniasis. 18,16 Den Leishmania spp.. parasiter kan vara konstruerad för att uttrycka eldfluga luciferas på en nivå som upptäcks in vivo med IVIS bildteknik. En av de stora fördelarna med denna metod är att det tillåter icke-invasiv visualisering av Leishmania spp.. inuti levande djur värd. Denna metod har till...

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">	
		
		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>D-Luciferin Potassium Salt </td>
<td>Reagent</td>
<td>Caliper LifeSciences (Formerly Xenogen)</td>
<td>122796</td>
<td>&nbsp;</td>
<tr>
<td>IVIS Imaging System 200 Series</td>
<td>Equipment</td>
<td>Caliper LifeSciences</td>
<td>&nbsp;</td>
<td>Other IVIS models that can be used include: Lumina II, Lumina XR, Kinetic, and Spectrum. </td>
</tr>		</tbody>
		</table>
		</div>

in vivo imaging of transgenic leishmania parasites in a live host

Distinkta arter av<em&gt; Leishmania,</em&gt; En protozo parasit i familjen<em&gt; Trypanosomatidae</em&gt;, Typiskt orsaka olika sjukdomar hos människan manifestationer. De vanligaste formerna av sjukdomen är visceral leishmaniasis (VL) och kutan leishmaniasis (CL). Musmodeller av leishmaniasis används allmänt, men kvantifiering av parasiten bördor under murina sjukdom kräver möss att avlivas vid olika tillfällen efter infektion. Parasit laster därefter mäts antingen med mikroskopi, begränsa utspädningen analys, eller qPCR amplifiering av parasitens DNA. Den<em&gt; In vivo</em&gt; Imaging System (IVIS) har ett integrerat programpaket som gör det möjligt att upptäcka en självlysande signal i samband med celler i levande organismer. Både för att minimera djurens användning och följa infektionen längden hos individer, in vivo-modeller för imaging<em&gt; Leishmania</em&gt; SPP. orsakar VL eller CL etablerades. Parasiter var konstruerad för att uttrycka luciferas, och dessa infördes i möss antingen intradermalt eller intravenöst. Kvantitativa mätningar av luciferas drivande mareld av transgena<em&gt; Leishmania</em&gt; Parasiter inom musen gjordes med hjälp IVIS. Individuella möss kan avbildas flera gånger under longitudinella studier, tillåter oss att bedöma mellan djur variation i den inledande experimentella parasit inokulat, och bedöma uppförökning av parasiter i mus vävnader. Parasiter upptäcks med hög känslighet vid kutan platser. Även om det är mycket troligt att signalen (fotoner / sekund / parasit) är lägre i djupare inre organ än huden, men kvantitativa jämförelser av signaler i ytlig respektive djup webbplatser som inte har gjorts. Det är möjligt att parasiten tal mellan kropp webbplatser inte direkt kan jämföras, men parasit belastningar i samma vävnader kan jämföras mellan möss. Exempel på en visceralizing arter (<em&gt; L. infantum chagasi</em&gt;) Och en art som orsakar kutan leishmaniasis (<em&gt; L. Mexicana</em&gt;) Visas. Det IVIS förfarande kan användas för att övervaka och analysera små djurmodeller av en mängd olika<em&gt; Leishmania</em&gt; Arter orsakar olika former av mänsklig leishmaniasis.

Watch this Scientific Journal Video about In vivo Imaging of Transgenic Leishmania Parasites in a Live Host at JoVE.com

In vivo Imaging of Transgenic Leishmania Parasites in a Live Host

En<em&gt; In vivo</em&gt; Imaging System används för att generera kvantitativa mätningar av murina infektion med Trypanosomatid protozo<em&gt; Leishmania</em&gt;. Detta är en icke-invasiv och icke-dödliga metod för att upptäcka parasiter uttrycka luciferas inom många vävnader under hela kroniska<em&gt; Leishmania</em&gt; SPP. infektion.

In vivo avbildning av Transgena Leishmania parasiter i ett Live-värd

Distinct species of Leishmania, a protozoan parasite of the family Trypanosomatidae, typically cause different human disease manifestations. The most ...

Research

JoVE Journal

Immunology and Infection

In vivo Imaging of Transgenic Leishmania Parasites in a Live Host

Non-invasive Imaging of Leukocyte Homing and Migration in vivo

Non-invasive Imaging of Leukocyte Homing and Migration in vivo

Icke-invasiv avbildning av Leukocyt Homing och migration In vivo</em

Two-photon (2P) microscopy is a high resolution imaging technique that has been broadly adapted by biologists.  The value of 2P microscopy is that it ...

Use of an Optical Trap for Study of Host-Pathogen Interactions for Dynamic Live Cell Imaging

Användning av en optisk fälla för studier av Host-patogen interaktioner för Dynamic levande cell imaging

Dynamic live cell imaging allows direct visualization of real-time interactions between cells of the immune system1, 2; however, the lack of spatial and ...

Induction of Graft-versus-host Disease and In Vivo T Cell Monitoring Using an MHC-matched Murine Model

Induction of Graft-versus-host Disease and In Vivo T Cell Monitoring Using an MHC-matched Murine Model

Induktion av transplantat-mot-värd-sjukdom och<em&gt; In Vivo</em&gt; T cell övervakning med en MHC-matchade musmodell

Graft-versus-host disease (GVHD) is the limiting barrier to the broad use of bone marrow transplant as a curative therapy for a variety of hematological ...

Human Placental and Decidual Organ Cultures to Study Infections at the Maternal-fetal Interface

Human placenta och decidual Organ kulturer att studera Infektioner hos moder och foster gränssnitt

The placenta shows a large degree of interspecies anatomic variability. To best understand biology and pathophysiology of the human placenta, it is ...

Quantification of Intracellular Growth Inside Macrophages is a Fast and Reliable Method for Assessing the Virulence of Leishmania Parasites

Quantification of Intracellular Growth Inside Macrophages is a Fast and Reliable Method for Assessing the Virulence of Leishmania Parasites

Kvantifiering av intracellulära tillväxt inuti makrofager är en snabb och tillförlitlig metod för att bedöma virulens Leishmania parasiter

The lifecycle of Leishmania, the causative agent of leishmaniasis, alternates between promastigote and amastigote stages inside the insect and vertebrate ...

In Vivo Imaging of Reactive Oxygen Species in a Murine Wound Model

In Vivo Imaging of Reactive Oxygen Species in a Murine Wound Model

In Vivo Avbildning av reaktiva syreradikaler i en murin sår modell

The generation of reactive oxygen species (ROS) is a hallmark of inflammatory processes, but in excess, oxidative stress is widely implicated in various ...

Human Colonoid Monolayers to Study Interactions Between Pathogens, Commensals, and Host Intestinal Epithelium

Mänskliga Colonoid enskiktslager att studera interaktioner mellan patogener, förknippas och värd Intestinal epitel

Human 3-dimensional (3D) enteroid or colonoid cultures derived from crypt base stem cells are currently the most advanced ex vivo model of the intestinal ...

In Vivo Infection with Leishmania amazonensis to Evaluate Parasite Virulence in Mice

In Vivo Infection with Leishmania amazonensis to Evaluate Parasite Virulence in Mice

In Vivo Infektion med Leishmania amazonensis att utvärdera parasit virulens i möss

Leishmania spp. are protozoan parasites that cause leishmaniases, diseases that present a wide spectrum of clinical manifestations from cutaneous to ...

Live Imaging of Chemokine Receptors in Zebrafish Neutrophils During Wound Responses

Levande avbildning av kemokinreceptorer i zebrafiskneutrofiler under sårsvar

Leukocyte guidance by chemical gradients is essential for immune responses. Neutrophils are the first cells to be recruited to sites of tissue damage ...

Recurrent Escherichia coli Urinary Tract Infection Triggered by Gardnerella vaginalis Bladder Exposure in Mice

Recurrent Escherichia coli Urinary Tract Infection Triggered by Gardnerella vaginalis Bladder Exposure in Mice

Återkommande Escherichia coli urinvägsinfektion utlöst av Gardnerella vaginalis urinblåsans exponering hos möss

Recurrent urinary tract infections (rUTI) caused by uropathogenic Escherichia coli (UPEC) are common and costly. Previous articles describing models of ...