In:
Journal of Immunology Research, Hindawi Limited, Vol. 2016 ( 2016), p. 1-11
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 (r Lr IL-10) using a heterologous expression system and its biophysical and functional characterization. High yield (~70 mg/L) of soluble r Lr IL-10 was obtained at shake flask level. The r Lr IL-10 was found to exist as a dimer. Far-UV CD spectroscopy showed presence of predominantly alpha helices. The tertiary structure of the purified r Lr IL-10 was verified by fluorescence spectroscopy. Two-dimensional gel analysis revealed the presence of six isoforms of the r Lr IL-10. The r Lr IL-10 was biologically active and its administration significantly reduced serum proinflammatory cytokines, namely, interleukin 1 β , TNF α , and IL-8, and augmented the NKEF transcript levels in spleen of L. rohita . Anti-inflammatory role of the r Lr IL-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.
Type of Medium:
Online Resource
ISSN:
2314-8861
,
2314-7156
DOI:
10.1155/2016/3962596
Language:
English
Publisher:
Hindawi Limited
Publication Date:
2016
detail.hit.zdb_id:
2817541-4
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