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Structural and Functional Characterization of Recombinant Interleukin-10 from Indian Major Carp Labeo rohita.


ABSTRACT: Interleukin-10, an important regulator of both the innate and adaptive immune systems, is a multifunctional major cytokine. Though it is one of the major cytokines, IL-10 from the Indian major carp, Labeo rohita, has not yet been characterized. In the present study, we report large scale production and purification of biologically active recombinant IL-10 of L. rohita (rLrIL-10) using a heterologous expression system and its biophysical and functional characterization. High yield (~70?mg/L) of soluble rLrIL-10 was obtained at shake flask level. The rLrIL-10 was found to exist as a dimer. Far-UV CD spectroscopy showed presence of predominantly alpha helices. The tertiary structure of the purified rLrIL-10 was verified by fluorescence spectroscopy. Two-dimensional gel analysis revealed the presence of six isoforms of the rLrIL-10. The rLrIL-10 was biologically active and its administration significantly reduced serum proinflammatory cytokines, namely, interleukin 1?, TNF?, and IL-8, and augmented the NKEF transcript levels in spleen of L. rohita. Anti-inflammatory role of the rLrIL-10 was further established by inhibition of phagocytosis using NBT reduction assay in vitro. The data indicate that the dimeric alpha helical structure and function of IL-10 of L. rohita as a key regulator of anti-inflammatory response have remained conserved during evolution.

SUBMITTER: Karan S 

PROVIDER: S-EPMC5023823 | biostudies-literature | 2016

REPOSITORIES: biostudies-literature

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Structural and Functional Characterization of Recombinant Interleukin-10 from Indian Major Carp <i>Labeo rohita</i>.

Karan Sweta S   Dash Pujarini P   Kaushik Himani H   Sahoo Pramoda K PK   Garg Lalit C LC   Dixit Aparna A  

Journal of immunology research 20160901


Interleukin-10, an important regulator of both the innate and adaptive immune systems, is a multifunctional major cytokine. Though it is one of the major cytokines, IL-10 from the Indian major carp, <i>Labeo rohita,</i> has not yet been characterized. In the present study, we report large scale production and purification of biologically active recombinant IL-10 of <i>L. rohita</i> (r<i>Lr</i>IL-10) using a heterologous expression system and its biophysical and functional characterization. High  ...[more]

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