Open Access

ARTICLE

Continuous large-scale production of the cytokine CXCL8 from a novel porcine cell line

Junsuke Shirai¹, Kikumi Ogihara², Ai Masumoto³, Kazuki Morioka¹, Yuko Naya², Yoshinori Tsuchiya¹, Yuichi Yokomizo⁴

1 Exotic Diseases Research Unit, National Institute of Animal Health, 6-20-1 Jyosui-honcho, Kodaira, Tokyo 187-0022, Japan
2 Department of Environmental Pathology, Azabu University, 1-17-71 Fuchinobe, Sagamihara, Kanagawa 229-8501, Japan
3 Department of Veterinary Science, Tokyo University of Agriculture and Technology, Fuchu, Tokyo 183-8509, Japan
4 Department of Immunology Research, National Institute of Animal Health, 3-1-5 Kannodai, Tsukuba, Ibaraki 305-0856, Japan

* Corresponding Author: J. Shirai,

European Cytokine Network 2007, 18(1), 10-17. https://doi.org/10.1684/ecn.2007.0081

Abstract

Cytokine production from two unstimulated porcine cell lines (SL-24 and SK-L) was examined using porcine cytokine detection ELISA kits and RT-PCR. Porcine IL-1α, IL-6, and CXCL8 were detected in all samples examined. In particular, the SL-24 cell line (derived from bone marrow cells of a malignant lymphoma-affected pig), produced large amounts of porcine CXCL8. Flow cytometer analysis showed the cell line to be strongly CD44 positive, and was therefore considered to be of monocyte or macrophage origin. Porcine CXCL8 production was greatest (83.86 ± 32.33 ng/mL) at six days post-cultivation. The SK-L cell line (derived from porcine kidney) also produced CXCL8, but production was less than 1.5 ng/mL. Porcine CXCL8 from the SL-24 cell line, induced chemotactic activity in porcine neutrophils, while the production of CXCL8 from the SL-24 cell line was inhibited by dexamethasone, which suggests that the mechanism of CXCL8 production is related to an NF-jB binding site. The production of CXCL8 from the SL-24 cell line was enhanced by the addition of recombinant porcine IL-15, which is the first reported observation of such CXCL8 production. Cloning of the SL-24 cell line by limited dilution revealed two types of cells present in the starting population. One cell type, designated as long-form cells (LC), produced large amounts of CXCL8, while the other, designated short-form cells (SC), produced small amounts of the cytokine. The LC cells were adapted to grow in serum-free medium in which they produced large amounts of CXCL8. The large-scale production of porcine CXCL8 from the SF-24 cell line will be of value in determining the mechanism of cytokine production and as a source of naturally produced porcine CXCL8.

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