Rabbit Anti-Human CTSW (a.a. 22-376) Polyclonal antibody

Oaxaca cheese is a Mexican pasta filata-type cheese, and can be considered as a potential vehicle for delivering probiotic cells. Their processing conditions are harsher than for most other cheeses. Thus, the challenge is to provide an adequate protection to the probiotic cells in order that they reach in sufficient numbers the sites for exerting their functionality. Lactobacillus plantarum cells were entrapped in double emulsions (DE) (aguamiel, Ag, or sweet whey, SW, as inner aqueous phase) and incorporated during cheese manufacture to obtain cheese treatments CTAg and CTSW. A control cheese (CTControl) was made with non-encapsulated free cells. Log CFU g(-1) was of 6.81 +/- 0.08, 8.15 +/- 0.17 and 822 +/- 0.09 for CTControl, CTSW and CTAg, respectively, immediately after CT's manufacture. After melting, Log CPU g(-1) dropped down by 2.18 +/- 0.17, 1.42 +/- 0.15 and 1.94 +/- 0.12 for CTControl, CTSw and CTAg, respectively. The exposure of CTAg and CTSw to simulated gastrointestinal conditions (pH of 2.3 and bile salts) did not affect significantly cells viability (6.32 +/- 0.13 and 6.02 +/- 0.1 Log CFU g(-1), respectively), but CTControl exhibited a significant decrease (p<0.05) to 3.36 +/- 0.01 Log CFU g(-1). The inclusion of cells in DE is an effective method for protecting their viability against harsh processing and usage conditions. (C) 2014 Elsevier Ltd. All rights reserved.

Human cathepsinW(CtsW) is a cysteine protease, which was identified in a genome-wide RNA interference (RNAi) screen to be required for influenza A virus (IAV) replication. In this study, we show that reducing the levels of expression of CtsW reduces viral titers for different subtypes of IAV, and we map the target step of CtsW requirement to viral entry. Using a set of small interfering RNAs (siRNAs) targeting CtsW, we demonstrate that knockdown of CtsW results in a decrease of IAV nucleoprotein accumulation in the nuclei of infected cells at 3 h postinfection. Assays specific for the individual stages of IAV entry further show that attachment, internalization, and early endosomal trafficking are not affected by CtsW knockdown. However, we detected impaired escape of viral particles from late endosomes in CtsW knockdown cells. Moreover, fusion analysis with a dual-labeled influenza virus revealed a significant reduction in fusion events, with no detectable impact on endosomal pH, suggesting that CtsW is required at the stage of viral fusion. The defect in IAV entry upon CtsW knockdown could be rescued by ectopic expression of wild-type CtsW but not by the expression of a catalytically inactive mutant of CtsW, suggesting that the proteolytic activity of CtsW is required for successful entry of IAV. Our results establish CtsW as an important host factor for entry of IAV into target cells and suggest that CtsW could be a promising target for the development of future antiviral drugs. IMPORTANCE Increasing levels of resistance of influenza viruses to available antiviral drugs have been observed. Development of novel treatment options is therefore of high priority. In parallel to the classical approach of targeting viral enzymes, a novel strategy is pursued: cell-dependent factors of the virus are identified with the aim of developing small-molecule inhibitors against a cellular target that the virus relies on. For influenza A virus, several genome-wide RNA interference (RNAi) screens revealed hundreds of potential cellular targets. However, we have only limited knowledge on how these factors support virus replication, which would be required for drug development. We have characterized cathepsin W, one of the candidate factors, and found that cathepsinWis required for escape of influenza virus from the late endosome. Importantly, this required the proteolytic activity of cathepsin W. We therefore suggest that cathepsin W could be a target for future host cell-directed antiviral therapies.