Human CASP3 ELISA Matched Antibody Pair

Objectives: Neutrophil apoptosis is mandatory for resolving inflammation and is regulated by expression of pro- and anti-apoptotic genes. We studied neutrophils isolated from patients with granulomatosis with polyangiitis (GPA) to investigate apoptosis alterations and to identify transcriptional and circulating factors affecting this process. Method: We enrolled 36 patients (18 in active stage, 18 in remission) and 18 healthy controls. Circulating levels of tumour necrosis factor-? (TNF-?), granulocyte?macrophage colony-stimulating factor (GM-CSF), macrophage migration inhibitory factor, plasminogen activator inhibitor-1, interferon-?, vascular cell adhesion molecule-1, intercellular adhesion molecule-1, platelet endothelial cell adhesion molecule-1, soluble Fas (sFas), sFas ligand, survivin, and pentraxin-3 (PTX3) were evaluated by enzyme-linked immunosorbent assay/Luminex; circulating apoptotic neutrophils by flow cytometry; and apoptosis-related gene transcripts by real-time polymerase chain reaction. Results: Patients had decreased fractions of circulating apoptotic neutrophils and delayed neutrophil apoptosis was present in vitro. Circulating levels of TNF-?, GM-CSF, sFas, and PTX3 were higher in GPA. Delayed neutrophil apoptosis was accompanied by decreased mRNA of pro-apoptotic genes and transcription factors (DIABLO, PMAIP1, BAX, CASP3, CASP7, RUNX3, E2F1, TP53) and increased anti-apoptotic CFLAR and BCL2A1 mRNA. TNF-? and sFas levels correlated with circulating apoptotic neutrophils and expression of apoptosis genes. Stimulation with TNF-? of neutrophils from controls significantly down-regulated E2F1 and CASP3 expression. Conclusions: Circulating neutrophils in GPA have anti-apoptotic phenotype involving both intrinsic and extrinsic pathways of apoptosis. This is accompanied by increased levels of circulating pro-survival factors (GM-CSF, TNF-?, sFas), independent of disease activity. Anti-apoptotic phenotype of neutrophils in GPA is reproduced by exposure to low concentrations of TNF-?.

Today, tumor therapy and its therapeutic efficiency evaluation are conducted separately, and current imaging techniques cannot evaluate tumor-therapeutic effects in real time. Therefore, it is of great importance to develop highly efficient theranostic strategies which are able to evaluate their tumor-therapeutic effects in real time. In this work, by rational design of a small molecular near-infrared probe Cys(StBu)-Asp-Glu-Val-Asp-Lys(Cypate)-CBT (Cy-CBT) and using a CBT-Cys click condensation reaction, we facilely prepare an intelligent nanoparticle Cy-CBT-NP which is able to evaluate its photothermal therapy (PTT) efficiency on tumors by fluorescence "Turn-On". Fluorescence of Cy-CBT-NP is quenched and photothermal responsive. Upon caspase 3 (Casp3) cleavage of its DEVD substrates, Cy-CBT-NP disassembles to turn the fluorescence "On", which in turn evaluates the PTT efficiency of the nanoparticle on cells and tumors in real time. We envision that our smart strategy could be applied for PTT and real-time evaluation of the therapeutic efficiency of solid tumors in the near future.