Stable transfection of Eimeria tenella: Constitutive expression of the YFP-YFP molecule throughout the life cycle

Yan, Wenchao; Liu, Xianyong; Shi, Tuanyuan; Hao, Lili; Tomley, Fiona M.; Suo, Xun

doi:10.1016/j.ijpara.2008.06.013

Cited by 59 publications

(62 citation statements)

References 32 publications

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“…Our previous work showed that the reproductivity of transgenic E. tenella expressing exogenous proteins was substantially decreased relative to the WT parasite (18). We examined the infectivity and immunogenicity of our transgenic E. tenella in chickens.…”

Section: Resultsmentioning

confidence: 99%

“…Electroporation was performed in a Gene Pulser Xcell Electroporation System (Bio-Rad, Hercules, CA) at 2000 V, 25 mF, and a pulse time of 0.3-0.4 ms. The sporozoites were allowed to recover for 20 min at room temperature and then inoculated to the ileocecal opening of 4-d-old chickens for in vivo stable transfection (18).…”

Section: Transfection and Selection Of Transgenic Parasitesmentioning

confidence: 99%

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Transgenic Eimeria tenella Expressing Enhanced Yellow Fluorescent Protein Targeted to Different Cellular Compartments Stimulated Dichotomic Immune Responses in Chickens

Huang

Zou

et al. 2011

The Journal of Immunology

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Eimeria tenella, one of the seven species of chicken coccidia, elicits protective immunity against challenge infection with both homologous and heterologous strains. We endeavor to use recombinant E. tenella as a vaccine vehicle for expressing and delivering pathogen Ags and investigate immune responses against these foreign Ags. In this study, two lines of transgenic E. tenella expressing a model Ag, enhanced yellow fluorescent protein (EYFP), targeted to the micronemes and to the cytoplasm of the recombinant parasites were constructed to study the impact of Ag compartmentalization on immunogenicity. The MTT assay, intracellular cytokine staining, and real-time PCR were performed to detect the EYFP-specific proliferation and effector functions of splenic lymphocytes of immunized chickens. ELISA was used to measure anti-EYFP IgG and IgA responses. The results showed that both lines of transgenic parasites stimulated EYFP-specific lymphocyte proliferation and IFN-γ expression in CD4 and CD8 T cells, whereas a higher level of Ag-specific lymphocyte proliferation was elicited by the transgenic line expressing microneme-targeted EYFP. Furthermore, this line stimulated stronger IgA response than the one expressing cytoplasm-targeted EYFP after the second immunization. Our findings are encouraging for further investigation of the effect of Ag compartmentalization in transgenic Eimeria on immunogenicity and for the development of a eukaryotic vaccine vector using genetically modified Apicomplexa parasites.

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Section: Resultsmentioning

confidence: 99%

Section: Transfection and Selection Of Transgenic Parasitesmentioning

confidence: 99%

Transgenic Eimeria tenella Expressing Enhanced Yellow Fluorescent Protein Targeted to Different Cellular Compartments Stimulated Dichotomic Immune Responses in Chickens

Huang

Zou

et al. 2011

The Journal of Immunology

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“…However, the transcription of cyst wall proteins-CWP1, CPW2, and CWP3-and of glucosamine-6-phosphate isomerase (i.e., GFAT equivalent) in encysting Giardia lamblia is synchronized (21) and under the control of a single transcription factor, gMyb2, itself upregulated during encystation (39). The identification of a macrogamete-specific transcription factor and promoter (i.e., that might be controlling the transcription of glycosylation enzymes) could help identify other sexual stage proteins of E. tenella and have applications (33,44). Glycosylation is utilized by other eukaryotes for a multitude of functions, including protein folding, targeting, recognition, and adhesion (26).…”

Section: Discussionmentioning

confidence: 99%

The Glycosylation Pathway of Eimeria tenella Is Upregulated during Gametocyte Development and May Play a Role in Oocyst Wall Formation

et al. 2010

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Sexual-stage glycoproteins of Eimeria are important components of the oocyst wall, a structure that ensures the efficient transmission of these and related parasites. In this study, the primary enzyme in the glycosylation pathway of Eimeria tenella, glucosamine:fructose-6-phosphate aminotransferase (EtGFAT), has been characterized as a macrogamete-specific protein. Although the transcription of EtGFAT was observed early in macrogamete development, protein expression was restricted to mature macrogametes, prior to their conversion into unsporulated oocysts. Genes coding for three other enzymes required for N-acetylgalactosamine (GalNAc) synthesis were also transcribed during E. tenella macrogamete development. Gene transcription of the enzyme responsible for the O-linked transfer of GalNAc to proteins, EtGalNAc-T, was upregulated primarily in unsporulated oocyst stages, and accordingly, a significant increase in GalNAc levels was observed in E. tenella gametocytes and oocysts. Gam56 and Gam82, two well-characterized glycoproteins of Eimeria macrogametes and the oocyst wall, contain high levels of GalNAc and represent probable targets of GalNAc O linkage. It appears that the glycosylation pathway, specifically relating to the formation of GalNAc O links, is dramatically upregulated in E. tenella sexual stages and may play a role in directing a number of macrogamete proteins to the developing oocyst wall.

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“…The different strands of work to identify protective antigens are now being brought together through the pioneering advances of Eimeria transfection technologies by Fiona Tomley and colleagues (for example, Yan et al, 2009). These powerful methodologies continue to take to a new level the ability of coccidiologists to evaluate the role and utility of critical gene products, including: those that are involved with parasite motility and invasion of the host cell; those contributing to the structure and function of sub-cellular organelles, especially rhoptries and micronemes; those responsible for traits such as virulence and host specificity and lifecycle differentiation through the different asexual and sexual stages; and those involved in induction of protective immune responses.…”

Section: Setting the Scenementioning

confidence: 99%

The long view: a selective review of 40 years of coccidiosis research

Shirley¹,

Lillehoj

2012

Avian Pathology

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This selective review of 40 years of coccidiosis research is one of a number on important diseases of poultry to celebrate the 40th anniversary of the birth of Avian Pathology, the journal of the World Veterinary Poultry Association, and is written for the non-specialist. The intention is to provide a flavour of the field problems and intellectual challenges, with emphasis in the areas of immunology and vaccinology that drove research in the 1970s, and to reflect on research progress since.

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Stable transfection of Eimeria tenella: Constitutive expression of the YFP-YFP molecule throughout the life cycle

Cited by 59 publications

References 32 publications

Transgenic Eimeria tenella Expressing Enhanced Yellow Fluorescent Protein Targeted to Different Cellular Compartments Stimulated Dichotomic Immune Responses in Chickens

Transgenic Eimeria tenella Expressing Enhanced Yellow Fluorescent Protein Targeted to Different Cellular Compartments Stimulated Dichotomic Immune Responses in Chickens

The Glycosylation Pathway of Eimeria tenella Is Upregulated during Gametocyte Development and May Play a Role in Oocyst Wall Formation

The long view: a selective review of 40 years of coccidiosis research

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