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Proteins induced by recombinant equine interferon-beta 1 within equine peripheral blood mononuclear cells and
H Heinz1, J Marquardt, H J Schuberth
1Immunology Unit, Veterinary School, Hannover, Germany.
Abstract:
Peripheral blood mononuclear cells (PBMC) and polymorphonuclear neutrophilic granulocytes (PMN) as well as embryonic equine dermal fibroblasts and the equine fibroblast line E. Derm which were used as controls, were treated with recombinant equine interferon-beta 1 (rEqIFN-beta 1) in vitro which induced the expression of different proteins in these cells. A 74 kDa protein was induced in PBMC and an 82 kDa protein was additionally found in the equine fibroblast E. Derm cell line following treatment with rEqFN-beta 1. Both proteins reacted with anti-mouse and anti-human Mx protein antisera in immunoblot tests. The 74 kDa and perhaps the 82 kDa components may thus represent equine 'Mxanalogous proteins'. The 74 kDa protein was only detected in PBMC of ten out of 20 horses examined. The induction of Mx protein in the horse by Type 1 interferon may therefore resemble that in the mouse, where Mx protein is involved in selective resistance to influenza virus. The influence of rEqIFN-beta 1 on protein expression in equine PBMC and PMN was monitored by metabolic labeling and 2-D gel electrophoresis. Proteins of 82, 74, 58 and 40 kDa were induced in PBMC following exposure to rEqIFN-beta 1. A constitutively expressed 35 kDa protein, however, was no longer demonstrable upon treatment with interferon. None of the proteins induced within PBMC was found in highly purified PMN treated with interferon. PMN exposed to rEqIFN-beta 1 synthesized four proteins in the range of 25 to 27 kDa. These proteins have not been described in interferon-treated PMN of any other species.
Insights
Recombinant equine interferon-beta 1 (rEqIFN-beta 1) induces Mx-analogous proteins in horse cells. These proteins, particularly in peripheral blood mononuclear cells (PBMC), show potential for equine antiviral resistance.
Area of Science:
- Immunology
- Virology
- Protein Biochemistry
Background:
- Interferons are crucial for antiviral defense.
- Equine interferon-beta 1 (IFN-beta 1) is a type 1 interferon with potential therapeutic applications.
- Understanding interferon-induced protein expression in equine cells is vital for antiviral strategies.
Purpose of the Study:
- To investigate the protein expression profile in equine cells upon treatment with recombinant equine interferon-beta 1 (rEqIFN-beta 1).
- To identify and characterize interferon-induced proteins, including potential Mx-analogous proteins, in equine peripheral blood mononuclear cells (PBMC) and polymorphonuclear neutrophilic granulocytes (PMN).
- To compare protein induction patterns between different equine cell types and relate them to known interferon responses in other species.
Main Methods:
- In vitro treatment of equine cells (PBMC, PMN, dermal fibroblasts) with rEqIFN-beta 1.
- Metabolic labeling and 2-D gel electrophoresis to monitor protein expression changes.
- Immunoblot analysis using anti-Mx protein antisera to detect Mx-analogous proteins.
Main Results:
- rEqIFN-beta 1 induced several proteins in equine PBMC, including 82, 74, 58, and 40 kDa proteins, and suppressed a 35 kDa protein.
- A 74 kDa protein, and potentially an 82 kDa protein in fibroblasts, reacted with anti-Mx antisera, suggesting they are equine Mx-analogous proteins.
- PMN treated with rEqIFN-beta 1 synthesized unique 25-27 kDa proteins not observed in other species' interferon-treated PMN.
Conclusions:
- Equine interferon-beta 1 induces the expression of Mx-analogous proteins in PBMC, similar to findings in mice, suggesting a role in antiviral resistance.
- Protein induction patterns differ significantly between equine PBMC and PMN following rEqIFN-beta 1 treatment.
- The identification of novel interferon-induced proteins in equine PMN warrants further investigation into their function and species-specific roles in antiviral immunity.