Dataset on characterization of recombinant interleukin-23α, IL-12p40 and IL-23 complex protein, which activates JAK-STAT signaling pathway in chicken cell lines using immunocytochemical staining

The data herein is related to the research article entitled “Functional analyses of the interaction of chicken interleukin 23 subunit p19 with IL-12 subunit p40 to form the IL-23 complex” [1] where we demonstrated that the chicken interleukin (IL)-23α, IL-12p40, and IL-23 complex regulates Th1, Th17, and Treg cytokine production through heterodimer receptors as well as a homodimer receptor consisting of IL-12Rβ1 and IL-23R, and activates the JAK/STAT signaling pathways. Here, we evaluated the effects of the recombinant chicken IL-23α, IL-12p40, and IL-23 complex protein on cell proliferation and nitric oxide (NO) production in chicken macrophage (HD11) and CU91 T cell lines. In addition, the expression of IL-6, IL-17A, and interferon-γ mRNA were upregulated in vivo and in vitro. Moreover, treatment with the chicken IL-23α, IL-12p40, and IL-23 complex activated phosphorylation of tyrosine and serine residues in JAK2, STAT1, TYK2, and SOCS1 in chicken cell lines.


a b s t r a c t
The data herein is related to the research article entitled "Functional analyses of the interaction of chicken interleukin 23 subunit p19 with IL-12 subunit p40 to form the IL-23 complex" [1] where we demonstrated that the chicken interleukin (IL)-23α, IL-12p40, and IL-23 complex regulates Th1, Th17, and Treg cytokine production through heterodimer receptors as well as a homodimer receptor consisting of IL-12Rβ1 and IL-23R, and activates the JAK/ STAT signaling pathways. Here, we evaluated the effects of the recombinant chicken IL-23α, IL-12p40, and IL-23 complex protein on cell proliferation and nitric oxide (NO) production in chicken macrophage (HD11) and CU91 T cell lines. In addition, the expression of IL-6, IL-17A, and interferon-γ mRNA were upregulated in vivo and in vitro. Moreover, treatment with the chicken To analyze recombinant proteins, western blot analysis using specific a HRP-anti-His (C-Term) antibody Nitric oxide content and cell proliferation were measured as described [2,3] To analyze signaling protein expression, immunocytochemical analysis using specific bodies were used [4] Data format Analyzed Experimental factors Recombinant protein was produced in E. coli and purified using HisPur™ Cobalt Resin; HD11 and OU2 cell lines were treated with recombinant protein as described [5] Experimental features

Anseong, Republic of Korea
Data accessibility Data are provided with this article

Value of the data
The data is valuable for the expression of pro-inflammatory molecules in chicken cell lines treated with DMSO and LPS and various tissues of chicken following S. enteritidis infection.
The data is a contribution to the effect of chicken IL-23α, IL-12p40, IL-12p40 þ IL-23α, and IL-23 complex protein to cell proliferation and production of reactive oxygen species in the form of NO in chicken cell lines.
The data provided the expression of JAK-STAT signaling molecules by chicken IL-23α, IL-12p40, IL-12p40 þ IL-23α, and IL-23 complex protein stimulation in chicken cell lines.

Data
The dataset in this article provides additional information to Ref. [1], where we demonstrated that the chicken IL-23α, IL-12p40, and IL-23 complex activated multiple signaling pathways through heterodimer receptors, as well as a homodimer receptor consisting of IL-12Rβ1 and IL-23R, and induced Th1, Th17, and Treg cytokine production. In this dataset, we provided the nucleotide and amino acid sequences of the chicken IL-23α coding region (Fig. 1A). These proteins were observed as single bands at 34 kDa (ChIL-23α), 48 kDa (ChIL-12p40-G10S3 linker), and 68 kDa (ChIL-23 complex) (Fig. 1B) by western blotting using the horseradish peroxidase (HRP)-anti-His (C-Term) antibody (Invitrogen, Carlsbad, CA, USA). The size was larger than predicted because of the presence of 3 epitope tags (polyhistidine, S-protein, and thioredoxin) in the recombinant protein. Moreover, data provides information about the effect of chicken IL-23α, IL-12p40, and IL-23 complex protein on cell proliferation and production of reactive oxygen species in the form of NO in both cell lines (Fig. 1C-D). In addition to the dataset, we present the expression of IL-6, IL-17A, and interferon (IFN)-γ mRNA in chicken cell lines treated with dimethyl sulfoxide (DMSO) and lipopolysaccharide (LPS) and in various tissues of the chicken following Salmonella Enteritidis infection (Fig. 2). Finally, immunocytochemical analysis showed that the expression of JAK-STAT signaling molecules by IL-23α, IL-12p40, and IL-23 complex stimulation in the chicken cell lines (Figs. 3 and 4).

Experimental design, materials, and methods
The experimental design is described in details in [1]. Detailed information about the methodology for chicken infection, samples collection, cell stimulation, recombinant protein production and   purification, quantitative real-time PCR (qRT-PCR), bioactivity assay, immunocytochemical, western blot and statistical analysis can be found elsewhere [1].

Acknowledgments
This research was supported by a National Research Foundation grant (2015R1D1A1A09059345) of the Republic of Korea.

Transparency document. Supporting information
Transparency data associated with this article can be found in the online version at https://doi.org/ 10.1016/j.dib.2017.12.008. Fig. 4. Immunocytochemical analysis of STAT1 (above) and SOCS1 (below) signaling proteins in HD11 cell lines (left) and CU91 T cell line (right). Both untreated and recombinant protein treated cells were incubated with primary antibody, Alexa Fluor ® 488 Goat Anti-Rabbit IgG (Hþ L) secondary antibody (green color) and DAPI (blue color) stained. Scale bar 25 µm.