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Development of a Cell Marker ELISA for the Detection of Goose T Cell Surface CD8α Molecules

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Abstract

CD8 molecule is a key marker on T cell surface and is connected with the antigen recognition and activation of T lymphocytes. In order to provide a detection method for quantifying goose CD8α expression, this study raised the protein and antibody for goose CD8α and developed a feasible cell marker enzyme-linked immunoabsorbent assay (ELISA) method. Recombinant protein of the extracellular region gene of goCD8α was expressed in prokaryotic expression system, and specific polyclonal antibodies for goCD8α were raised and purified, which was further confirmed by Western-blot, immunofluorescence assay (IFA), and immunohistochemistry (IHC). A cell marker ELISA was established and optimized to detect the change of goCD8α expression between goose parvovirus (GPV)-infected and mock-infected goose peripheral blood mononuclear cells (PBMCs), which is consistent with our previously results of real-time quantitative PCR (qPCR). Cell marker ELISA can provide a new method to detect goCD8α in protein level and in a sensitive, specific, and simple way. This may provide a convenient and novel method for the detection of goCD8α expression.

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Abbreviations

CTL:

Cytotoxic T lymphocytes

ELISA:

Enzyme-linked immunosorbent assay

GPV:

Goose parvovirus

IFA:

Immunofluorescence assay

IHC:

Immunohistochemistry

IPTG:

Isopropyl β-d-thiogalactopyranoside

PBMCs:

Peripheral blood mononuclear cells

qPCR:

Quantitative PCR

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

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Acknowledgments

This work was funded by grants from the National Natural Science Foundation of China (31201891), The Ph.D. Programs Foundation of Ministry of Education of China (20125103120012), the Innovative Research Team Program in Education Department of Sichuan Province (2013TD0015), the National Science and Technology Support Program (2015BAD12B05), Integration and Demonstration of Key Technologies for Duck Industrial in Sichuan Province (2014NZ0030), and China Agricultural Research System (CARS-43-8).

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Authors and Affiliations

  1. Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, No. 211 Huimin Road, Wenjiang District, Chengdu City, Sichuan Province, 611130, China

    Beibei Cheng, Wei Zhang, Shun Chen, Mingshu Wang, Renyong Jia, Mafeng Liu, Kunfeng Sun, Qiao Yang, Ying Wu & Anchun Cheng

  2. Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China

    Shun Chen, Mingshu Wang, Renyong Jia, Dekang Zhu, Kunfeng Sun, Qiao Yang, Ying Wu, Xiaoyue Chen & Anchun Cheng

  3. Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China

    Shun Chen, Mingshu Wang, Renyong Jia, Dekang Zhu, Kunfeng Sun, Qiao Yang, Ying Wu, Xiaoyue Chen & Anchun Cheng

Authors
  1. Beibei Cheng
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  2. Wei Zhang
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  3. Shun Chen
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  4. Mingshu Wang
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  5. Renyong Jia
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  6. Dekang Zhu
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  7. Mafeng Liu
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  8. Kunfeng Sun
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  9. Qiao Yang
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  10. Ying Wu
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  11. Xiaoyue Chen
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  12. Anchun Cheng
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Corresponding authors

Correspondence to Shun Chen or Anchun Cheng.

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The authors declare that they have no competing interests.

Additional information

Beibei Cheng, Wei Zhang, and Shun Chen contributed equally as co-first authors of this work.

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Cheng, B., Zhang, W., Chen, S. et al. Development of a Cell Marker ELISA for the Detection of Goose T Cell Surface CD8α Molecules. Appl Biochem Biotechnol 179, 531–544 (2016). https://doi.org/10.1007/s12010-016-2011-1

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  • DOI: https://doi.org/10.1007/s12010-016-2011-1

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