ABSTRACT
The exotoxins TcdA and TcdB are the major virulence factors of Clostridium difficile. Circulating neutralizing antitoxin antibodies are protective in C. difficile infection (CDI), as demonstrated, in part, by the protective effects of actoxumab and bezlotoxumab, which bind to and neutralize TcdA and TcdB, respectively. The question of how systemic IgG antibodies neutralize toxins in the gut lumen remains unresolved, although it has been suggested that the Fc receptor FcRn may be involved in active antibody transport across the gut epithelium. In this study, we demonstrated that genetic ablation of FcRn and excess irrelevant human IgG have no impact on actoxumab-bezlotoxumab-mediated protection in murine and hamster models of CDI, suggesting that Fc-dependent transport of antibodies across the gut wall is not required for efficacy. Tissue distribution studies in hamsters suggest, rather, that the transport of antibodies depends on toxin-induced damage to the gut lining. In an in vitro two-dimensional culture system that mimics the architecture of the intestinal mucosal epithelium, toxins on the apical side of epithelial cell monolayers are neutralized by basolateral antibodies, and antibody transport across the cell layer is dramatically increased upon addition of toxin to the apical side. Similar data were obtained with F(ab')2 fragments, which lack an Fc domain, consistent with FcRn-independent paracellular, rather than transcellular, transport of antibodies. Kinetic studies show that initial damage caused by apical toxin is required for efficient neutralization by basolateral antibodies. These data may represent a general mechanism of humoral response-mediated protection against enteric pathogens.
Subject(s)
Antibodies, Bacterial/immunology , Antibodies, Neutralizing/immunology , Antitoxins/immunology , Bacterial Proteins/toxicity , Bacterial Toxins/toxicity , Enterotoxins/toxicity , Animals , Antibodies, Bacterial/metabolism , Antibodies, Bacterial/therapeutic use , Antibodies, Neutralizing/metabolism , Antibodies, Neutralizing/therapeutic use , Antitoxins/metabolism , Antitoxins/therapeutic use , Bacterial Proteins/immunology , Bacterial Toxins/immunology , Clostridioides difficile/immunology , Clostridium Infections/therapy , Disease Models, Animal , Enterotoxins/immunology , Female , Histocompatibility Antigens Class I , Immunization, Passive , Immunoglobulin G/immunology , Immunoglobulin G/metabolism , Immunoglobulin G/therapeutic use , Male , Mesocricetus , Mice, Inbred C57BL , Mice, Knockout , Organ Culture Techniques , Receptors, Fc/deficiencyABSTRACT
Farnesyl-protein transferase (FPTase) is a critical enzyme that participates in the post-translational modification of the Ras protein. Inhibitors of this enzyme have the potential of being novel anticancer agents for tumors in which the ras oncogene is found mutated and contributes to cell transformation. Continued screening of natural product extracts led to the isolation of kampanols, which are novel and specific inhibitors of FPTase. The most active kampanols exhibited IC50 values between 7 to 13 microM against human recombinant FPTase. The isolation, structure determination, and biological activity of these compounds are described.
Subject(s)
Alkyl and Aryl Transferases/antagonists & inhibitors , Antineoplastic Agents/chemistry , Enzyme Inhibitors/chemistry , Terpenes/chemistry , Antineoplastic Agents/isolation & purification , Antineoplastic Agents/pharmacology , Enzyme Inhibitors/isolation & purification , Enzyme Inhibitors/pharmacology , Genes, ras , Humans , Magnetic Resonance Spectroscopy , Models, Molecular , Molecular Conformation , Molecular Structure , Recombinant Proteins/antagonists & inhibitors , Terpenes/isolation & purification , Terpenes/pharmacologyABSTRACT
A novel fungal metabolite, apicidin [cyclo(N-O-methyl-L-tryptophanyl-L -isoleucinyl-D-pipecolinyl-L-2-amino-8-oxodecanoyl)], that exhibits potent, broad spectrum antiprotozoal activity in vitro against Apicomplexan parasites has been identified. It is also orally and parenterally active in vivo against Plasmodium berghei malaria in mice. Many Apicomplexan parasites cause serious, life-threatening human and animal diseases, such as malaria, cryptosporidiosis, toxoplasmosis, and coccidiosis, and new therapeutic agents are urgently needed. Apicidin's antiparasitic activity appears to be due to low nanomolar inhibition of Apicomplexan histone deacetylase (HDA), which induces hyperacetylation of histones in treated parasites. The acetylation-deacetylation of histones is a thought to play a central role in transcriptional control in eukaryotic cells. Other known HDA inhibitors were also evaluated and found to possess antiparasitic activity, suggesting that HDA is an attractive target for the development of novel antiparasitic agents.
Subject(s)
Antiprotozoal Agents/pharmacology , Enzyme Inhibitors/pharmacology , Eukaryota/drug effects , Histone Deacetylase Inhibitors , Malaria/drug therapy , Peptides, Cyclic/pharmacology , Plasmodium berghei , Animals , Eimeria tenella/drug effects , Female , Humans , Kinetics , Mice , Mice, Inbred BALB C , Neospora/drug effects , Peptides, Cyclic/therapeutic use , Plasmodium falciparum/drug effects , Protein Binding , Protozoan Infections/drug therapy , Structure-Activity Relationship , Toxoplasma/drug effectsABSTRACT
Two new zaragozic acids, D and D2, have been isolated from the keratinophilic fungus Amauroascus niger. Zaragozic acids D [4] and D2 [5] are related to the previously described zaragozic acids A [1], B [2], and C [3] and are potent inhibitors of squalene synthase. Furthermore, all the zaragozic acids (A, B, C, D, and D2) are also active against farnesyl transferase. Zaragozic acids D and D2 inhibit farnesyl transferase with IC50 values of 100 nM, while zaragozic acids A and B are less potent.
