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Molecular Medicine

, Volume 19, Issue 1, pp 65–71 | Cite as

Effect of Recombinant α1-Antitrypsin Fc-Fused (AAT-Fc) Protein on the Inhibition of Inflammatory Cytokine Production and Streptozotocin-Induced Diabetes

  • Siyoung Lee
  • Youngmin Lee
  • Kwangwon Hong
  • Jaewoo Hong
  • Suyoung Bae
  • Jida Choi
  • Hyunjhung Jhun
  • Areum Kwak
  • Eunsom Kim
  • Seunghyun Jo
  • Charles A. Dinarello
  • Soohyun Kim
Research Article

Abstract

α1-Antitrypsin (AAT) is a member of the serine proteinase inhibitor family that impedes the enzymatic activity of serine proteinases, including human neutrophil elastase, cathepsin G and neutrophil proteinase 3. Here, we expressed recombinant AAT by fusing the intact AAT gene to the constant region of IgG1 to generate soluble recombinant AAT-Fc protein. The recombinant AAT-Fc protein was produced in Chinese hamster ovary (CHO) cells and purified using mini-protein A affinity chromatography. Recombinant AAT-Fc protein was tested for antiinflammatory function and AAT-Fc sufficiently suppressed tumor necrosis factor (TNF)-α-induced interleukin (IL)-6 in human peripheral blood mononuclear cells (PBMCs) and inhibited cytokine-induced TNFα by different cytokines in mouse macrophage Raw 264.7 cells. However, AAT-Fc failed to suppress lipopolysaccharide-induced cytokine production in both PBMCs and macrophages. In addition, our data showed that AAT-Fc blocks the development of hyperglycemia in a streptozotocin-induced mouse model of diabetes. Interestingly, we also found that plasma-derived AAT specifically inhibited the enzymatic activity of elastase but that AAT-Fc had no inhibitory effect on elastase activity.

Notes

Acknowledgments

This work was supported by National Research Foundation of Korea grant funded by the Korea government (MEST: 2012R1A2A1A01001791 and WCU: R33-2008-000-10022-0). S Kim received support from the Konkuk University research support program.

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Authors and Affiliations

  • Siyoung Lee
    • 1
  • Youngmin Lee
    • 2
  • Kwangwon Hong
    • 1
  • Jaewoo Hong
    • 1
  • Suyoung Bae
    • 1
  • Jida Choi
    • 1
  • Hyunjhung Jhun
    • 1
  • Areum Kwak
    • 1
  • Eunsom Kim
    • 1
  • Seunghyun Jo
    • 1
  • Charles A. Dinarello
    • 3
  • Soohyun Kim
    • 1
  1. 1.Laboratory of Cytokine Immunology, Department of Biomedical Sciences and TechnologyKonkuk UniversitySeoulKorea
  2. 2.Department of Medicine, Pusan Paik Hospital, College of MedicineInje UniversityBusanKorea
  3. 3.Department of MedicineUniversity of Colorado Denver, AuroraDenverUSA

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