Molecular determinants and mechanism for antibody cocktail preventing SARS-CoV-2 escape.

Antibody cocktails represent a promising approach to prevent SARS-CoV-2 escape. The determinants for selecting antibody combinations and the mechanism that antibody cocktails prevent viral escape remain unclear. We compared the critical residues in the receptor-binding domain (RBD) used by multiple neutralizing antibodies and cocktails and identified a combination of two antibodies CoV2-06 and CoV2-14 for preventing viral escape. The two antibodies simultaneously bind to non-overlapping epitopes and independently compete for receptor binding. SARS-CoV-2 rapidly escapes from individual antibodies by generating resistant mutations in vitro, but it doesn't escape from the cocktail due to stronger mutational constraints on RBD-ACE2 interaction and RBD protein folding requirements. We also identified a conserved neutralizing epitope shared between SARS-CoV-2 and SARS-CoV for antibody CoV2-12. Treatments with CoV2-06 and CoV2-14 individually and in combination confer protection in mice. These findings provide insights for rational selection and mechanistic understanding of antibody cocktails as candidates for treating COVID-19.

Specificity of cobamide remodeling, uptake and utilization in Vibrio cholerae.

Cobamides are a group of compounds including vitamin B12 that can vary at the lower base position of the nucleotide loop. They are synthesized de novo by only a subset of prokaryotes, but some organisms encode partial biosynthesis pathways for converting one variant to another (remodeling) or completing biosynthesis from an intermediate (corrinoid salvaging). Here, we explore the cobamide specificity in Vibrio cholerae through examination of three natural variants representing major cobamide groups: commercially available cobalamin, and isolated pseudocobalamin and p-cresolylcobamide. We show that BtuB, the outer membrane corrinoid transporter, mediates the uptake of all three variants and the intermediate cobinamide. Our previous work suggested that V. cholerae could convert pseudocobalamin produced by cyanobacteria into cobalamin. In this work, cobamide specificity in V. cholerae is demonstrated by remodeling of pseudocobalamin and salvaging of cobinamide to produce cobalamin. Cobamide remodeling in V. cholerae is distinct from the canonical pathway requiring amidohydrolase CbiZ, and heterologous expression of V. cholerae CobS was sufficient for remodeling. Furthermore, function of V. cholerae cobamide-dependent methionine synthase MetH was robustly supported by cobalamin and p-cresolylcobamide, but not pseudocobalamin. Notably, the inability of V. cholerae to produce and utilize pseudocobalamin contrasts with enteric bacteria like Salmonella.

Reduction of Bacterial Pathogens and Potential Surrogates on the Surface of Almonds Using High-Intensity 405-Nanometer Light.

The disinfecting properties of high-intensity monochromatic blue light (MBL) were investigated against Escherichia coli O157:H7, Salmonella , and nonpathogenic bacteria inoculated onto the surface of almonds. MBL was generated from an array of narrow-band 405-nm light-emitting diodes. Almonds were inoculated with higher or lower levels (8 or 5 CFU/g) of pathogenic E. coli O157:H7 and Salmonella , as well as nonpathogenic E. coli K-12 and an avirulent strain of Salmonella Typhimurium, for evaluation as potential surrogates for their respective pathogens. Inoculated almonds were treated with MBL for 0, 1, 2, 4, 6, 8, and 10 min at a working distance of 7 cm. Simultaneous to treatment, cooling air was directed onto the almonds at a rate of 4 ft3/min (1.89 ×10-3 m3/s), sourced through a container of dry ice. An infrared camera was used to monitor the temperature readings after each run. For E. coli K-12, reductions of up to 1.85 or 1.63 log CFU/g were seen for higher and lower inoculum levels, respectively; reductions up to 2.44 and 1.44 log CFU/g were seen for E. coli O157:H7 (higher and lower inoculation levels, respectively). Attenuated Salmonella was reduced by up to 0.54 and 0.97 log CFU/g, whereas pathogenic Salmonella was reduced by up to 0.70 and 0.55 log CFU/g (higher and lower inoculation levels, respectively). Inoculation level did not significantly impact minimum effective treatment times, which ranged from 1 to 4 min. Temperatures remained below ambient throughout treatment, indicating that MBL is a nonthermal antimicrobial process. The nonpathogenic strains of E. coli and Salmonella each responded to MBL in a comparable manner to their pathogenic counterparts. These results suggest that these nonpathogenic strains may be useful in experiments with MBL in which a surrogate is required, and that MBL warrants further investigation as a potential antimicrobial treatment for low-moisture foods.