“Self-cleaving” 2A peptide from porcine teschovirus-1 mediates cleavage of dual fluorescent proteins in transgenic Eimeria tenella

Tang, Xinming; Liu, Xianyong; Tao, Geru; Qin, Mei; Yin, Guangwen; Suo, Jingxia; Suo, Xun

doi:10.1186/s13567-016-0351-z

Cited by 41 publications

(46 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…GC catalytic dead (TgGC E2987A-mCherry), 8. Skip peptide (Tang et al, 2016) separating the P-type ATPase and GC domains (TgGC P2A-mCherry), 9. Non-skip peptide control (TgGC P2A GP/AA -mCherry).…”

Section: Resultsmentioning

confidence: 99%

Essential cGMP Signaling in Toxoplasma Is Initiated by a Hybrid P-Type ATPase-Guanylate Cyclase

Brown

Sibley

2018

Cell Host & Microbe

101

View full text Add to dashboard Cite

Summary (150) Apicomplexan parasites rely on cyclic nucleotide-dependent kinases for host cell infection, yet the mechanisms that control their activation remain unknown. Here we show that an apically-localized guanylate cyclase (GC) controls microneme secretion and lytic growth in the model apicomplexan Toxoplasma gondii. Cell permeable cGMP reversed the block in microneme secretion seen in a knockdown of TgGC, linking its function to production of cGMP. TgGC possesses an N-terminal P-type-ATPase domain fused to a C-terminal heterodimeric guanylate cyclase domain, an architecture found only in Apicomplexa and related protists. Complementation with a panel of mutants revealed a critical requirement for the P-type ATPase domain for maximum GC function. We further demonstrate that knockdown of TgGC in vivo protects mice from lethal infection by blocking parasite expansion and dissemination. Collectively this work demonstrates that cGMP-mediated signaling in Toxoplasma relies on a multi-domain architecture, which may serve a conserved role in related parasites.

show abstract

Section: Resultsmentioning

confidence: 99%

Essential cGMP Signaling in Toxoplasma Is Initiated by a Hybrid P-Type ATPase-Guanylate Cyclase

Brown

Sibley

2018

Cell Host & Microbe

101

View full text Add to dashboard Cite

show abstract

“…Here, we used the regulatory sequences of E. tenella , which was shown to be reliable in a previous study on transfection of E. intestinalis (26) in a plasmid generated by Tang et al (10). This reliability might result from the functional conservation of E. tenella histone 4 and actin genes among these Eimeria species.…”

Section: Discussionmentioning

confidence: 99%

“…This reliability might result from the functional conservation of E. tenella histone 4 and actin genes among these Eimeria species. Additionally, Tang et al (10) provided an in-depth study on E. tenella demonstrating the “self-cleavage” peptide 2A from porcine teschovirus-1. We also found that it was efficient in our transgenic E. magna .…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Transgenic Eimeria magna Pérard, 1925 Displays Similar Parasitological Properties to the Wild-type Strain and Induces an Exogenous Protein-Specific Immune Response in Rabbits (Oryctolagus cuniculus L.)

et al. 2017

Self Cite

View full text Add to dashboard Cite

Rabbit coccidiosis causes great economic losses to world rabbitries. Little work has been done considering genetic manipulation on the etiological agents, rabbit Eimeria spp. In this study, we constructed a transgenic line of Eimeria magna (EmagER) expressing enhanced yellow fluorescent protein (EYFP) and red fluorescent protein (RFP) using regulatory sequences of Eimeria tenella and Toxoplasma gondii. We observed the life cycle of EmagER and confirmed that the transgenic parasites express exogenous proteins targeted to different cellular compartments throughout the entire life cycle. EYFP was expressed mainly in the nucleus and RFP both in the nucleus and cytoplasm. Then, coccidia-free, laboratory-reared 40-day-old rabbits were primarily infected with either EmagER or wild-type strain oocysts and challenged with the wild-type strain. EmagER showed similar reproductivity and immunogenicity to the wild-type strain. Finally, we examined the foreign protein-specific immune response elicited by EmagER. Rabbits were immunized with either transgenic or wild-type oocysts. Immune response against parasite-soluble antigen, EYFP and RFP in spleen, and mesenteric lymph nodes were detected by quantitative real-time PCR. The relative expression level of IFN-γ, IL-2, and TNF-α were higher in EmagER-immunized rabbits than wild-type parasites-immunized rabbits after stimulation with EYFP and RFP. Our study confirmed that a specific immune response was induced by the exogenous protein expressed by EmagER and favored future studies on application of transgenic rabbit coccidia as recombinant vaccine vectors.

show abstract

“…The first small 'self-cleaving' peptide F2A was identified in the foot-and-mouth disease virus, a member of the picornavirus group (35,36). Subsequently, '2A like' peptides from equine rhinitis A virus, P2A and T2A were identified, and their activities in proteolytic cleavage were certified in various in vitro and in vivo eukaryotic systems (35,36). Subsequently, the PLAC1-TCR gene was expressed in primary human CD8+T cells using a negative selection process using magnetic beads.…”

Section: Discussionmentioning

confidence: 99%

PLAC1-specific TCR-engineered T cells mediate antigen-specific antitumor effects in breast cancer

Liu

et al. 2018

Oncol Lett

View full text Add to dashboard Cite

Abstract. Placenta-specific 1 (PLAC1), a novel cancer-testis antigen (CTA), is expressed in a number of different human malignancies. It is frequently produced in breast cancer, serving a function in tumorigenesis. Adoptive immunotherapy using T cell receptor (TCR)-engineered T cells against CTA mediates objective tumor regression; however, to the best of our knowledge, targeting PLAC1 using engineered T cells has not yet been attempted. In the present study, the cDNAs encoding TCRα-and β-chains specific for human leukocyte antigen (HLA)-A * 0201-restricted PLAC1 were cloned from a cytotoxic T-lymphocyte, generated by in vitro by the stimulation of CD8+ T cells using autologous HLA-A2+ dendritic cells loaded with a PLAC1-specific peptide (p28-36, VLCSIDWFM). The TCRα/β-chains were linked by a 2A peptide linker (TCRα-Thosea asigna virus-TCRβ), and the constructs were cloned into the lentiviral vector, followed by transduction into human cytotoxic (CD8+) T cells. The efficiency of transduction was up to 25.16%, as detected by PLAC1 multimers. TCR-transduced CD8+ T cells, co-cultured with human non-metastatic breast cancer MCF-7 cells (PLAC1+, HLA-A2+) and triple-negative breast cancer MDAMB-231 cells (PLAC1+, HLA-A2+), produced interferon γ and tumor necrosis factor α, suggesting TCR activation. Furthermore, the PLAC1 TCR-transduced CD8+ T cells efficiently and specifically identified and annihilated the HLA-A2+/PLAC1+ breast cancer cell lines in a lactate dehydrogenase activity assay. Western blot analysis demonstrated that TCR transduction stimulated the production of mitogen-activated protein kinase signaling molecules, extracellular signal-regulated kinases 1/2 and nuclear factor-κB, through phosphoinositide 3-kinase γ-mediated phosphorylation of protein kinase B in CD8+ T cells. Xenograft mouse assays revealed that PLAC1 TCR-transduced CD8+T cells significantly delayed the tumor progression in mice-bearing breast cancer compared with normal saline or negative control-transduced groups. In conclusion, a novel HLA-A2-restricted and PLAC1-specific TCR was identified. The present study demonstrated PLAC1 to be a potential target for breast cancer treatment; and the usage of PLAC1-specific TCR-engineered T cells may be a novel strategy for PLAC1-positive breast cancer treatment.

show abstract

“Self-cleaving” 2A peptide from porcine teschovirus-1 mediates cleavage of dual fluorescent proteins in transgenic Eimeria tenella

Cited by 41 publications

References 26 publications

Essential cGMP Signaling in Toxoplasma Is Initiated by a Hybrid P-Type ATPase-Guanylate Cyclase

Essential cGMP Signaling in Toxoplasma Is Initiated by a Hybrid P-Type ATPase-Guanylate Cyclase

Transgenic Eimeria magna Pérard, 1925 Displays Similar Parasitological Properties to the Wild-type Strain and Induces an Exogenous Protein-Specific Immune Response in Rabbits (Oryctolagus cuniculus L.)

PLAC1-specific TCR-engineered T cells mediate antigen-specific antitumor effects in breast cancer

Contact Info

Product

Resources

About