Cyclophosphamide

The STAT3 inhibitor Stattic® was prepared and applied prior to vaccinia scarification as previously described [4 (link)]. Cyclophosphamide (Santa Cruz Biotech, Dallas, TX) was administered by intraperitoneal route starting with the day prior to vaccinia scarification and every 4 days after vaccinia infection as previously described [8 (link)]. Scarification using 10⁶ pfu ACAM-2000 was performed as previously described [4 (link)].

Cyclophosphamide was obtained from Santa Cruz Biotechnology, Inc. (California, USA). Interleukin-18 (IL-18; SL0400Ra), Interleukin-1β (IL-1β; SL0402Ra), Macrophage inflammatory protein2 (MIP2; SL0465Ra), cytokine-induced- neutrophil chemoattractant (CINC; SL1588Ra), Matrix metalloproteinase 1 (MMP1; SL0480Ra), NLR pyrin domain protein 3 (NLRP3; SL1497Ra) and caspase-1 (SL1601Ra) were determined using ELISA kits (Sunlong Biotech Co., Ltd, China). All other chemicals used in this study are of the highest grade commercially available.

Cyclophosphamide and protocatechuic acid were purchased from (Santa Cruz, CA, USA). Glutathione (GSH), malondialdehyde (MDA), nitric oxide (NO), and catalase colorimetric kits were determined using Biodiagnostic kit, Cairo, Egypt. Sirtuin 1 (SIRT1) was determined using ELISA kits procured from (SunRed Biotech Co., Ltd, China). Tumor necrosis factor alpha (TNF-α), Interleukin-17 (IL-17), nuclear factor kappa B (NF-κB), inhibitor of nuclear factor kappa beta kinase subunit beta (IKBKB), proxisome proliferator-activated receptor gamma (PPARγ), cyclooxegnase-2 (COX-2), protein kinase C (PKC), and myeloperoxidase (MPO) were determined using ELISA kits procured from Sunlong Biotech Co., Ltd, China.