Effect of recombinant α1-antitrypsin Fc-fused (AAT-Fc)protein on the inhibition of inflammatory cytokine production and streptozotocin-induced diabetes

Mol Med. 2013 May 20;19(1):65-71. doi: 10.2119/molmed.2012.00308.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anti-Inflammatory Agents / pharmacology*
  • Anti-Inflammatory Agents / therapeutic use
  • Blood Glucose / analysis
  • CHO Cells
  • Cell Line
  • Cells, Cultured
  • Cricetinae
  • Cricetulus
  • Cytokines / pharmacology
  • Diabetes Mellitus, Experimental / drug therapy
  • Diabetes Mellitus, Experimental / metabolism
  • Female
  • Humans
  • Immunoglobulin Fc Fragments / genetics
  • Immunoglobulin Fc Fragments / pharmacology
  • Immunoglobulin Fc Fragments / therapeutic use
  • Immunoglobulin G / genetics
  • Immunoglobulin G / pharmacology*
  • Immunoglobulin G / therapeutic use
  • Interleukin-6 / metabolism
  • Leukocytes, Mononuclear / drug effects
  • Leukocytes, Mononuclear / metabolism
  • Mice
  • Mice, Inbred BALB C
  • Pancreatic Elastase / metabolism
  • Recombinant Fusion Proteins / pharmacology*
  • Recombinant Fusion Proteins / therapeutic use
  • Tumor Necrosis Factor-alpha / metabolism
  • alpha 1-Antitrypsin / genetics
  • alpha 1-Antitrypsin / pharmacology*
  • alpha 1-Antitrypsin / therapeutic use

Substances

  • Anti-Inflammatory Agents
  • Blood Glucose
  • Cytokines
  • Immunoglobulin Fc Fragments
  • Immunoglobulin G
  • Interleukin-6
  • Recombinant Fusion Proteins
  • Tumor Necrosis Factor-alpha
  • alpha 1-Antitrypsin
  • Pancreatic Elastase