Name: parasite induced genetically driven autoimmune chagas heart disease in the chicken model
Uploaded: 2012-07-29T14:00:00
Duration: 9 min 23 s
Description: Watch this Scientific Journal Video about Parasite Induced Genetically Driven Autoimmune Chagas Heart Disease in the Chicken Model at JoVE.com

Somos gratos a Nancy R. Sturm, Departamento de Imunologia, Microbiologia e Biologia Molecular, David Geffen School of Medicine, Universidade da Califórnia em Los Angeles, para a leitura crítica do manuscrito. O Conselho Nacional de Pesquisa-CNPq, ea Fundação para o Desenvolvimento da Pesquisa-FAPDF, Brasil, apoiou o estudo. Agradecemos a ajuda técnica de Alessandro O. Souza, C. Maria Guimaro, Cordeiro Ciro, de Ana Cassia Rosa, Roseneide Alves e Rafael Andrade, da Universidade de Brasília, Brasil.

<ol>
	<li>Teixeira, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathogenesis+of+chagas'+disease:+parasite+persistence+and+autoimmunity.">Pathogenesis of chagas' disease: parasite persistence and autoimmunity.</a> CMR. 24, 592-630 (2011).</li><li>Teixeira, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chagas+disease.">Chagas disease.</a> Postg. Med. J. 82, 788-798 (2006).</li><li>Teixeira, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trypanosoma+cruzi+in+the+chicken+model:+Chagas-like+heart+disease+in+the+absence+of+parasitism.">Trypanosoma cruzi in the chicken model: Chagas-like heart disease in the absence of parasitism.</a> PLoS Negl. Trop. Dis. 5, e1000 (2011).</li><li>Hecht, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inheritance+of+DNA+transferred+from+American+trypanosomes+to+human+hosts.">Inheritance of DNA transferred from American trypanosomes to human hosts.</a> PLoS One. 5, e9181 (2010).</li><li>Xing, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Roles+of+the+ERK+MAPK+in+the+regulation+of+proinflammatory+and+apoptotic+responses+in+chicken+macrophages+infected+with+H9N2+avian+influenza+virus.">Roles of the ERK MAPK in the regulation of proinflammatory and apoptotic responses in chicken macrophages infected with H9N2 avian influenza virus.</a> J. Gen. Virol. 91, 343-351 (2010).</li><li>Kim, H. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NIK+and+IKKbeta+interdependence+in+NF-kappaB+signalling--flux+analysis+of+regulation+through+metabolites.">NIK and IKKbeta interdependence in NF-kappaB signalling--flux analysis of regulation through metabolites.</a> Biosystems. 99, 140-149 (2010).</li><li>Karakhanova, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ERK/p38+MAP-kinases+and+PI3K+are+involved+in+the+differential+regulation+of+B7-H1+expression+in+DC+subsets.">ERK/p38 MAP-kinases and PI3K are involved in the differential regulation of B7-H1 expression in DC subsets.</a> Eur. J. Immunol. 40, 254-266 (2010).</li><li>Finsterer, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+heart+in+human+dystrophinopathies.">The heart in human dystrophinopathies.</a> Cardiology. 99, 1-19 (2003).</li><li>Nitz, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heritable+integration+of+kDNA+minicircle+sequences+from+Trypanosoma+cruzi+into+the+avian+genome:+insights+into+human+Chagas+disease.">Heritable integration of kDNA minicircle sequences from Trypanosoma cruzi into the avian genome: insights into human Chagas disease.</a> Cell. 118, 175-186 (2004).</li><li>Simpson, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinetoplast+DNA+in+trypanosomid+flagellates.">Kinetoplast DNA in trypanosomid flagellates.</a> Int. Rev. Cytol. 99, 1-19 (1986).</li><li>Bonney, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heat-killed+Trypanosoma+cruzi+induces+acute+cardiac+damage+and+polyantigenic+autoimmunity.">Heat-killed Trypanosoma cruzi induces acute cardiac damage and polyantigenic autoimmunity.</a> PLoS One. 6, e14571 (2011).</li><li>Moser, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+Trypanosoma+cruzi+by+DNA+amplification+using+the+polymerase+chain+reaction.">Detection of Trypanosoma cruzi by DNA amplification using the polymerase chain reaction.</a> J. Clin. Microbiol. 27, 1477-1482 (1989).</li><li>Sturm, N. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sensitive+detection+and+schizodeme+classification+of+Trypanosoma+cruzi+cells+by+amplification+of+kinetoplast+minicircle+DNA+sequences:+use+in+diagnosis+of+Chagas'+disease.">Sensitive detection and schizodeme classification of Trypanosoma cruzi cells by amplification of kinetoplast minicircle DNA sequences: use in diagnosis of Chagas' disease.</a> Mol. Biochem. Parasitol. 33, 205-214 (1989).</li><li>Teixeira, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evolution+and+pathology+in+chagas+disease--a+review.">Evolution and pathology in chagas disease--a review.</a> Mem. Inst. Oswaldo Cruz. 101, 463-491 (2006).</li><li>Yurchenko, V. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structure+of+Leishmania+minicircle+kinetoplast+DNA+classes.">Structure of Leishmania minicircle kinetoplast DNA classes.</a> J. Clin. Microbiol. 37, 1656-1657 (1999).</li><li>Simpson, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+genomic+organization+of+guide+RNA+genes+in+kinetoplastid+protozoa:+several+conundrums+and+their+solutions.">The genomic organization of guide RNA genes in kinetoplastid protozoa: several conundrums and their solutions.</a> Mol. Biochem. Parasitol. 86, 133-141 (1997).</li><li>Thomas, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+non-universal+transcription+factor?+The+Leishmania+tarentolae+TATA+box-binding+protein+LtTBP+associates+with+a+subset+of+promoters.">A non-universal transcription factor? The Leishmania tarentolae TATA box-binding protein LtTBP associates with a subset of promoters.</a> Int J. Parasitol. 36, 1217-1226 (2006).</li></ol>

Não há conflitos de interesse declarados.

In contrast to mammals susceptible to life-long T. cruzi infections, chickens are refractory to T. cruzi infection. The major advantage of the chicken model system is the elimination of the infection early in the development of the embryonic immune system. Thus, the only parasite DNA remaining within the chicken body is integrated at several loci.
Use of the optimal quantity of virulent T. cruzi trypomastigotes to inoculate fertile egg is the critical step towards obtaining integration of the kDNA minicircles into the chicken embryo genome. The rate of live chicks hatching from eggs inoculated with 100 trypomastigotes is four-fold higher than that obtained with 500 parasites. Care should be taken to inoculate the parasites suspension in 10 &mu;L of culture medium into the egg air chamber. There should be no leakage of egg white. Under optimal conditions, the intracellular parasitic infection takes place within a few hours after incubation and parasite multiplication inside the host cells proceeds for one week; thereafter the infection is eliminated by the innate immunity. The kDNA integration requires a living infection, and the inoculation of naked minicircles into early embryo chicken eggs does not result in integration. The kDNA-positive embryos and controls should be housed under controlled conditions at 37.5 &deg;C and 65% humidity. The chicks are kept in cages for two weeks at 33 &deg;C room temperature. Thereafter the chickens are kept in cages on suspended racks separated by 1.5 meter width aisles in a room at 22 &deg;C with filtered air and positive pressure under constant exhaustion to secure animal welfare conditions. The adults are fed chicken-chow and drink potable running water to achieve full growth and maturity, laying eggs at five months of age. Maintenance of hygienic procedures are essential for reproducibility of results when working with T. cruzi inoculated into fertile chicken eggs.
In the chicken model system the T. cruzi infections are eradicated after development of the immune system in the early period of embryo growth. Additionally to being infection-free, the chicks that hatch from T. cruzi inoculated eggs, in lack of specific antibodies, are tolerant to the parasite antigens. The rejection of the target heart cells by cytotoxic lymphocytes (minimal rejection unit, Figure 3) is seen in the kDNA-mutated chickens showing genotype modifications and breakdown of immunological surveillance 3. The genotypically modified T-cells present accelerated rejection of self tissues in the body. The main lesion site is the heart, which is a hallmark of Chagas disease. The passage from a physiologic (surveillance) to a physiopathologic state is seen in the kDNA-mutated chicken showing clonally proliferation of cytotoxic lymphocytes 3.
This transkingdom model system shows a parasite-induced, genetically-driven autoimmune disease (Figure 6), stemming from genome modifications by the T. cruzi kDNA minicircle integrations. These modifications are not seen in chicks hatched from Lb-inoculated eggs. 
This phenomenon suggests that experimental treatment of the inflammatory autoimmune cardiomyopathy in kDNA-mutated chickens may require drug suppression of bone marrow progenitor of specific T-cell phenotype infiltrating the myocardium, and transplantation of histocompatible healthy bone marrow to prevent the rejection of self-tissue.

<table border="1"><tbody><tr><td> Nome do reagente </td><td> Companhia </td><td> Número de catálogo </td><td> Comentários </td></tr><tr><td> Taq DNA polimerase recombinante </td><td> Invitrogen </td><td> 11615-010 </td><td></td></tr><tr><td> Platinum Taq DNA polimerase </td><td> Invitrogen </td><td> 10966-030 </td><td></td></tr><tr><td> Aleatória Primers DNA Labeling Sistema </td><td> Invitrogen </td><td> 18187-013 </td><td></td></tr><tr><td> Eco RI </td><td> Invitrogen </td><td> 15202-021 </td><td></td></tr><tr><td> Mbo I </td><td> Invitrogen </td><td> 15248-016 </td><td></td></tr><tr><td> dNTP Set, 100mM Solutions </td><td&gt; GE Healthcare </td><td> 28-4065-51 </td><td></td></tr><tr><td> Amersham Hybond - N + - Cat n. </td><td> GE Healthcare </td><td> RPN303B </td><td></td></tr><tr><td> PlasmidPrep Mini spin kit </td><td> GE Healthcare </td><td> 28-9042-70 </td><td></td></tr><tr><td> Nsi I </td><td> Sigma-Aldrich </td><td> R5884 1KU </td><td></td></tr><tr><td> DNA, de sódio de esperma de peixe Sal </td><td> Amresco </td><td> 0644-10G </td><td></td></tr><tr><td> De ratinho anti-galinha Bu-1b </td><td> SouthernBiotech </td><td> 8370-02 </td><td></td></tr><tr><td> De ratinho anti-CD45 de galinha </td><td> SouthernBiotech </td><td> 8270-08 </td><td></td></tr><tr><td>Rato anti-galinha TCRγδ </td><td> SouthernBiotech </td><td> 8230-08 </td><td></td></tr><tr><td> De ratinho anti-CD8a galinha </td><td> SouthernBiotech </td><td> 9220-02 </td><td></td></tr><tr><td> Rato monócitos anti-galinha / macrófagos </td><td> SouthernBiotech </td><td> 8420-02 </td><td></td></tr><tr><td> MyCycle Termocycler </td><td> Bio-Rad Laboratories </td><td> 580BR 5501 </td><td></td></tr></tbody></table>

parasite induced genetically driven autoimmune chagas heart disease in the chicken model

As infecções agudas do Trypanosoma cruzi adquiridos na infância parecem assintomáticas, mas aproximadamente um terço dos casos cronicamente infectados apresentam a doença de Chagas até três décadas ou mais tarde. Autoimunidade e persistência parasita estão competindo teorias para explicar a patogênese da doença de Chagas 1, 2. Para papéis distintos desempenhados por parasita persistência e auto-imunidade na doença de Chagas nos inocular o T. cruzi na câmara de ar de ovos fertilizados. O sistema imunológico maduro frango é uma barreira biológica apertado contra T. cruzi e que a infecção é erradicada sobre o desenvolvimento da sua sistema imunitário, até ao final da primeira semana de crescimento 3. Os filhotes são parasita-livre em eclosão, mas eles mantêm integrados parasita mitocondrial DNA do cinetoplasto (kDNA) minicírculos dentro de seu genoma que são transferidos aos seus descendentes. Documentação da integração kDNA minicírculo no genoma de galinha foi obtido por um targeted principal TAIL-PCR, hibridização do Sul, clonagem, sequenciamento e 3, 4. O kDNA minicírculos integrações ruptura quadros de leitura aberta para a transcrição e factores do sistema imunitário, fosfatase (GTPase), adenilato ciclase e fosforilases (PKC, NF-kappa B activador, PI-3K) associado com a fisiologia da célula, o crescimento, diferenciação e 3, 5 - 7, e as funções de outros genes. Miocardite grave devido a rejeição de fibras de coração do alvo por efectores linfócitos citotóxicos é visto nas galinhas kDNA mutados, mostrando uma cardiomiopatia inflamatória semelhante à observada na doença de Chagas humana. Notavelmente, a insuficiência cardíaca e fraqueza muscular esquelética estão presentes em frangos adultos com kDNA ruptura do gene da distrofina no cromossoma 1 8. Similares alterações genotípicas estão associados com a destruição do tecido realizado por efectores CD45 +, CD8γδ + e CD8a linfócitos. Assim, esta protozoose pode induzir geneticamente orientada doença auto-imune.

Watch this Scientific Journal Video about Parasite Induced Genetically Driven Autoimmune Chagas Heart Disease in the Chicken Model at JoVE.com

Parasite Induced Genetically Driven Autoimmune Chagas Heart Disease in the Chicken Model

A inoculação de<em&gt; Trypanosoma cruzi</em&gt; Em ovos férteis para incubação anteriores torna o parasita minicírculos do kDNA integração no genoma de células de embriões. Cruzamento revela a transferência vertical das mutações à progenitura. O kDNA integra em regiões codificantes em vários cromossomos e as galinhas morrer com uma doença cardíaca inflamatória auto-imune.

Parasite Induzida geneticamente guiadas Doenças Auto-Imunes do Coração Chagas no Modelo de Frango

The Trypanosoma cruzi acute infections acquired in infancy and childhood seem asymptomatic, but approximately one third of the chronically infected cases ...

Research

JoVE Journal

Immunology and Infection

Induction of Experimental Autoimmune Hypophysitis in SJL Mice

Indução de hipofisite Auto-Imune Experimental em Ratos SJL

Autoimmune hypophysitis can be reproduced experimentally by the injection of pituitary proteins mixed with an adjuvant into susceptible mice1. Mouse ...

Imunologia e Infecção

Myelin Oligodendrocyte Glycoprotein (MOG35-55) Induced Experimental Autoimmune Encephalomyelitis (EAE) in C57BL/6 Mice

Myelin Oligodendrocyte Glycoprotein MOG35-55 Induced Experimental Autoimmune Encephalomyelitis EAE in C57BL/6 Mice

Mielina Oligodendrocyte glicoproteína (MOG<sub&gt; 35-55</sub&gt;) Induzida Encefalomielite Auto-imune Experimental (EAE) em murganhos C57BL / 6

Multiple sclerosis is a chronic neuroinflammatory demyelinating disorder of the central nervous system with a strong neurodegenerative component. While ...

Induction of Experimental Autoimmune Encephalomyelitis in Mice and Evaluation of the Disease-dependent Distribution of Immune Cells in Various Tissues

Indução de encefalomielite autoimune experimental em camundongos e avaliação da distribuição da doença dependente de células imunes em vários tecidos

Multiple sclerosis is presumed to be an inflammatory autoimmune disease, which is characterized by lesion formation in the central nervous system (CNS) ...

Sexual Transmission of American Trypanosomes from Males and Females to Naive Mates

Transmissão sexual de tripanosomas americanas de machos e fêmeas para companheiros de ingênua

American trypanosomiasis is transmitted to humans by triatomine bugs through the ingestion of contaminated food, by blood transfusions or accidently in ...

An Ex Vivo Chicken Primary Bursal-cell Culture Model to Study Infectious Bursal Disease Virus Pathogenesis

An Ex Vivo Chicken Primary Bursal-cell Culture Model to Study Infectious Bursal Disease Virus Pathogenesis

Um modelo de cultura primária de células bolsas serosas Ex Vivo frango para estudar a patogênese de doenças infecciosas bolsas serosas vírus

Infectious bursal disease virus (IBDV) is a birnavirus of economic importance to the poultry industry. The virus infects B cells, causing morbidity, ...

A Murine Model of Dengue Virus-induced Acute Viral Encephalitis-like Disease

Um modelo murino da doença da encefalite viral aguda induzida pelo vírus da dengue

Dengue virus (DENV), an arthropod-borne virus transmitted by mosquitoes, may cause the severe disease known as dengue hemorrhagic fever, which is ...

Induction of Drug-Induced, Autoimmune Hepatitis in BALB/c Mice for the Study of Its Pathogenic Mechanisms

Indução de Hepatite Autoimune Induzida por Drogas em CAMUNDONGOS BALB/c para o Estudo de Seus Mecanismos Patogênicos

Drug-induced autoimmune hepatitis (DIH) is the most common hepatic drug-induced hypersensitization process observed in approximately 9 to 12% of patients ...

Adoptive Immunotherapy of iNKT Cells in Glucose-6-Phosphate Isomerase (G6PI)-Induced RA Mice

Adoptive Immunotherapy of iNKT Cells in Glucose-6-Phosphate Isomerase G6PI-Induced RA Mice

Imunoterapia adotiva de células iNKT em Isomerase Isomerase Induzida de Glicose-6-Fosfato (G6PI)-Induzido RaMice

Rheumatoid arthritis (RA) is a complex chronic inflammatory autoimmune disease. The pathogenesis of the disease is related to invariant natural killer T ...

In Vitro Microfluidic Disease Model to Study Whole Blood-Endothelial Interactions and Blood Clot Dynamics in Real-Time

Modelo de doença microfluida in vitro para estudar interações endoteliais de sangue inteiro e dinâmica do coágulo sanguíneo em tempo real

The formation of blood clots involves complex interactions between endothelial cells, their underlying matrix, various blood cells, and proteins. The ...

Propagation of the Microsporidian Parasite Edhazardia aedis in Aedes aegypti Mosquitoes

Propagation of the Microsporidian Parasite Edhazardia aedis in Aedes aegypti Mosquitoes

Propagação do Parasita Microsporidiano Edhazardia aedis em Mosquitos Aedes aegypti

Edhazardia aedis is a microsporidian parasite of Aedes aegypti mosquitoes, a disease vector that transmits multiple arboviruses which cause millions of ...