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Peer-reviewedCC BY-NC 4.0Iranian Journal of Microbiology

Anti-inflammatory effects of recombinant TGF-arazyme protein on endotoxin-inflamed lung cell lines

  • Roohollah Zarei Koosha1
  • Raheleh Halabian1
  • Mahdi Fasihi-Ramandi2
  • Abbas Ali Imani Fooladi1
  1. 1Applied Microbiology Research Center, Biomedicine Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran
  2. 2Molecular Biology Research Center, Biomedicine Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran
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Abstract

Background and Objectives:: Pulmonary inflammation is central to the pathogenesis of various respiratory diseases, driving the need for novel therapeutics. Lipopolysaccharide (LPS) is a potent inflammatory trigger that induces proinflammatory cytokine synthesis via distinct signaling cascades. The objectives of this study were to evaluate the cytotoxic and anti-inflammatory effects of recombinant TGF-Arazyme fusion protein on LPS-stimulated human lung A549 and MRC-5 cell lines.

Materials and Methods:: A gene construct fusing Arazyme with a non-mitogenic TGFα fragment was expressed, purified via Ni-NTA chromatography, and validated enzymatically. Cytotoxicity was assessed by MTT assay. Inflammatory cytokine levels (TNF-α, IL-1β, IL-6, IL-10) were quantified by RT-PCR and ELISA. Anti-inflammatory efficacy was further validated in a murine (BALB/c) air pouch model infected with E. coli.

Results:: At 20 μg/mL, the chimeric protein showed no significant toxicity in MRC-5 cells but dose-dependently reduced A549 viability (p<0.01). Co-treatment with LPS and the chimeric protein significantly downregulated proinflammatory cytokines (TNF-α, IL-1β, IL-6) at both mRNA and protein levels (p<0.01, p<0.001), while increasing IL-10 (p<0.05). In the air pouch model, TGF-Arazyme reduced leukocyte counts by 48.3% (p<0.01), comparable to dexamethasone (51.2%).

Conclusion:: TGF-Arazyme exhibits both anti-inflammatory and selective cytotoxic properties, positioning it as a promising candidate for inflammatory lung diseases and lung cancer.

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Cite

BibTeXRIS
BibTeX
@article{koosha2026antiinflammatory,
  title = {{Anti-inflammatory effects of recombinant TGF-arazyme protein on endotoxin-inflamed lung cell lines}},
  author = {Roohollah Zarei Koosha and Raheleh Halabian and Mahdi Fasihi-Ramandi and Abbas Ali Imani Fooladi},
  journal = {Iranian Journal of Microbiology},
  year = {2026},
  month = nov,
  doi = {10.18502/ijm.v18i5.22871},
  url = {https://doi.org/10.18502/ijm.v18i5.22871},
  note = {CC BY-NC 4.0},
}
RIS
TY  - JOUR
TI  - Anti-inflammatory effects of recombinant TGF-arazyme protein on endotoxin-inflamed lung cell lines
AU  - Roohollah Zarei Koosha
AU  - Raheleh Halabian
AU  - Mahdi Fasihi-Ramandi
AU  - Abbas Ali Imani Fooladi
PY  - 2026
DA  - 2026/11/06
JO  - Iranian Journal of Microbiology
DO  - 10.18502/ijm.v18i5.22871
UR  - https://doi.org/10.18502/ijm.v18i5.22871
AB  - Background and Objectives:: Pulmonary inflammation is central to the pathogenesis of various respiratory diseases, driving the need for novel therapeutics. Lipopolysaccharide (LPS) is a potent inflammatory trigger that induces proinflammatory cytokine synthesis via distinct signaling cascades. The objectives of this study were to evaluate the cytotoxic and anti-inflammatory effects of recombinant TGF-Arazyme fusion protein on LPS-stimulated human lung A549 and MRC-5 cell lines.  Materials and Methods:: A gene construct fusing Arazyme with a non-mitogenic TGFα fragment was expressed, purified via Ni-NTA chromatography, and validated enzymatically. Cytotoxicity was assessed by MTT assay. Inflammatory cytokine levels (TNF-α, IL-1β, IL-6, IL-10) were quantified by RT-PCR and ELISA. Anti-inflammatory efficacy was further validated in a murine (BALB/c) air pouch model infected with E. coli.  Results:: At 20 μg/mL, the chimeric protein showed no significant toxicity in MRC-5 cells but dose-dependently reduced A549 viability (p<0.01). Co-treatment with LPS and the chimeric protein significantly downregulated proinflammatory cytokines (TNF-α, IL-1β, IL-6) at both mRNA and protein levels (p<0.01, p<0.001), while increasing IL-10 (p<0.05). In the air pouch model, TGF-Arazyme reduced leukocyte counts by 48.3% (p<0.01), comparable to dexamethasone (51.2%).  Conclusion:: TGF-Arazyme exhibits both anti-inflammatory and selective cytotoxic properties, positioning it as a promising candidate for inflammatory lung diseases and lung cancer.
SN  - 2008-3289
SN  - 2008-4447
ER  -