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

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 1β,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. � 2016 sweta karan et al.