Evaluation of interferon gamma (IFN-γ) of Labeo rohita as an immunomodulator: in vitro expression model
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Title |
Evaluation of interferon gamma (IFN-γ) of Labeo rohita as an immunomodulator: in vitro expression model
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Creator |
Bedekar MK, Soman P, Kole S, Anand D, Tripathi G, Makesh M, Rajendran KV
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Subject |
IFN-γ . Labeo rohita . Peripheral blood mononuclear cells . Poly I:C
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Description |
Not Available
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 polyinosinicpolycytidylic 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 Not Available |
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Date |
2022-06-18T05:51:12Z
2022-06-18T05:51:12Z 2018-07-31 |
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Type |
Research Paper
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Identifier |
Not Available
Not Available http://krishi.icar.gov.in/jspui/handle/123456789/72884 |
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Language |
English
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Relation |
Not Available;
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Publisher |
CrossMark
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