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Evaluation of interferon gamma (IFN-γ) of Labeo rohita as an immunomodulator: in vitro expression model

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Abstract

Interferon gamma (IFN-γ) or type II interferon is a cytokine that is critical for innate and adaptive immunity against viral and some bacterial and protozoal infections. The importance of IFN-γ in the immune system lies in its ability to inhibit viral replication directly and most importantly from its immunomodulatory effects. Previously, we successfully co-administered IFN-γ along with GAPDH gene of Edwardsiella tarda as bicistronic DNA vaccine in Labeo rohita. In order to ascertain the individual role of IFN-γ, the present study involves cloning and expression of 552-bp IFN-γ open-reading frame (ORF) of L. rohita in striped snakehead (SSN-1) cell line using eukaryotic expression vector system (pQE-TriSystem) followed by transfection in peripheral blood lymphocytes (PBMCs) to evaluate its immunomodulatory ability in comparison to polyinosinic-polycytidylic acid (Poly I:C)-treated PBMCs. The 18.7-kDa protein, expressed in the pQE-IFNγ-transfected SSN-1 cells, reacted with anti-His antibody in Western blot confirming it to be recombinant IFN-γ, whereas the relative expression of IFN-γ, iNOS, Mx, and IL-1β genes in PBMCs was quantified at 24 h and 48 h post treatment by qPCR. The comparative kinetics of all four genes showed significantly (p < 0.05) high upregulation pattern in both pQE-IFNγ-transfected cell group and Poly I:C-treated cell group demonstrating recombinant IFN-γ as an equally efficient inducer like Poly I:C. Thus, our in vitro experiment results highlight the immunomodulatory potential of recombinant IFN-γ as an analogue to synthetic Poly I:C which warranted future studies to further explore the potential of recombinant IFN-γ as an effective vaccine adjuvant against different microbial invasion.

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Acknowledgements

The authors thank the present Director Dr. Gopal Krishna and Ex-Director Dr. W S Lakra, ICAR-Central Institute of Fisheries Education, Mumbai, India, for providing necessary facilities and support for this study.

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Author notes

  1. Sajal Kole

    Present address: Department of Aqualife Medicine, Chonnam National University, Gwangju, Republic of Korea

Authors and Affiliations

  1. ICAR-Central Institute of Fisheries Education, Mumbai, India

    Megha Kadam Bedekar, Praveena Soman, Sajal Kole, Deepika Anand, Gayatri Tripathi & K. V. Rajendran

  2. ICAR-Central Institute of Brackishwater Aquaculture, Chennai, India

    M. Makesh

Authors
  1. Megha Kadam Bedekar
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  2. Praveena Soman
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  3. Sajal Kole
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  4. Deepika Anand
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  5. Gayatri Tripathi
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  6. M. Makesh
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  7. K. V. Rajendran
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Correspondence to Megha Kadam Bedekar.

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Highlights

• The 552-bp ORF of IFN-γ of L. rohita cloned in pQE-TriSystem expression vector.

• The recombinant IFN-γ positively expresses in SSN-1 cells and PBMCs.

• IFN-γ, iNOS, Mx, and IL-1β genes significantly upregulated in pQE-IFNγ-transfected cells.

• The recombinant IFN-γ was observed to be an equally efficient inducer like Poly I:C.

• Recombinant IFN-γ can be used as an immunomodulator in therapeutic treatment in fish.

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Bedekar, M.K., Soman, P., Kole, S. et al. Evaluation of interferon gamma (IFN-γ) of Labeo rohita as an immunomodulator: in vitro expression model. Aquacult Int 26, 1401–1413 (2018). https://doi.org/10.1007/s10499-018-0292-9

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  • DOI: https://doi.org/10.1007/s10499-018-0292-9

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