In vivo application of a small molecular weight antifungal protein of Penicillium chrysogenum (PAF)

Abstract

The small antifungal protein secreted by Penicillium chrysogenum (PAF) inhibits the growth of important zoo pathogenic filamentous fungi, including members of the Aspergillus family.It was shown previously that PAF has no toxic effects on mammalian cells in vitro. We carried out safety experiments by investigating the in vivo effects of PAF by inoculating adult C57/B16 mice with PAF intranasally. Animals were randomly divided into six groups and subjected to different PAF concentrations, up to 54 µg, either once a week for up to 8 weeks or once every day for two weeks. Animals neither died due to the treatment nor were any side effects observed. Histological examinations did not find any pathological reaction in the liver, in the mucous membrane of the nose, and in the lungs, even in the highest concentration used.Mass spectrometry revealed that ZZZ. The effect of the drug on the skin was examined in an irritative dermatitis model by measuring the thickness of the ears. This proved to be the same following PAF application as in control (23.8±9.2 vs. 22.5±5.0 µm, respectively) and significantly less than when treated with phorbol-12-myristate-13-acetate (PMA; 57.5±29.2 µm) used as positive control. Histological changes relative to control were present only in the case of PMA. Positron emission tomography was used to follow potential inflammation of the lungs. Neither the application of control saline nor that of PAF induced any inflammation while the positive control lipopolysaccharide did. Since no toxic effects of PAF were found either in intranasal application or in local external treatment, our result is the first step for introducing PAF as potential antifungal drug in human therapy.

A Penicillium chrysogenum-based expression system for the production of small, cysteine-rich antifungal proteins for structural and functional analyses

“…These natural proteins inhibit the growth of human-, animal- and plant-pathogenic moulds [8]. No detrimental effects on plant or on mammalian cells in vitro and in vivo could be observed for PAF [9, 10] and AFP [11, 12]. These findings strongly support their applicability as new antifungal drugs or their use for the development of new antifungal strategies.…”

Synthesis of PAF, an Antifungal Protein from P. chrysogenum, by Native Chemical Ligation: Native Disulfide Pattern and Fold Obtained upon Oxidative Refolding

“…6 It was previously observed that PAF can effectively inhibit the growth of numerous filamentous fungi causing membrane permeabilization, intracellular oxidative stress, and evoking programmed cell death through a G‐protein signal‐transduction pathway 7. PAF is harmless for mammals, as proven in our recent PET (positron emission tomography) experiments 8. Therefore, PAF is promising for the treatment of mycoses and fungal infections, especially against aspergillosis.…”

D19S Mutation of the Cationic, Cysteine-Rich Protein PAF: Novel Insights into Its Structural Dynamics, Thermal Unfolding and Antifungal Function

“…It is a prepro-protein which is processed before secretion and the mature PAF consists of 55 amino acids (Fig 1A) [4]. It specifically inhibits the growth of opportunistic human- and plant-pathogens, such as Aspergillus fumigatus and Botrytis cinerea , but is inactive against mammalian cells both in vitro and in vivo [5,6]. In the course of our intensive studies to understand the mechanistic action of PAF, we have investigated its solution structure in great detail [7,8].…”