Single domain antibodies against enteric pathogen virulence factors are active as curli fiber fusions on probiotic E. coli Nissle 1917

PLoS Pathogens · 2022 · 18 citations

• Microbiology
• Biology

Enteric microbial pathogens, including Escherichia coli, Shigella and Cryptosporidium species, take a particularly heavy toll in low-income countries and are highly associated with infant mortality. We describe here a means to display anti-infective agents on the surface of a probiotic bacterium. Because of their stability and versatility, VHHs, the variable domains of camelid heavy-chain-only antibodies, have potential as components of novel agents to treat or prevent enteric infectious disease. We isolated and characterized VHHs targeting several enteropathogenic E. coli (EPEC) virulence factors: flagellin (Fla), which is required for bacterial motility and promotes colonization; both intimin and the translocated intimin receptor (Tir), which together play key roles in attachment to enterocytes; and E. coli secreted protein A (EspA), an essential component of the type III secretion system (T3SS) that is required for virulence. Several VHHs that recognize Fla, intimin, or Tir blocked function in vitro. The probiotic strain E. coli Nissle 1917 (EcN) produces on the bacterial surface curli fibers, which are the major proteinaceous component of E. coli biofilms. A subset of Fla-, intimin-, or Tir-binding VHHs, as well as VHHs that recognize either a T3SS of another important bacterial pathogen (Shigella flexneri), a soluble bacterial toxin (Shiga toxin or Clostridioides difficile toxin TcdA), or a major surface antigen of an important eukaryotic pathogen (Cryptosporidium parvum) were fused to CsgA, the major curli fiber subunit. Scanning electron micrographs indicated CsgA-VHH fusions were assembled into curli fibers on the EcN surface, and Congo Red binding indicated that these recombinant curli fibers were produced at high levels. Ectopic production of these VHHs conferred on EcN the cognate binding activity and, in the case of anti-Shiga toxin, was neutralizing. Taken together, these results demonstrate the potential of the curli-based pathogen sequestration strategy described herein and contribute to the development of novel VHH-based gut therapeutics.

Evaluating the concordance of functional MRI-based language lateralization and Wada testing in epilepsy patients: A single-center analysis

Interventional Neuroradiology · 2022 · 7 citations

• Audiology
• Medicine
• Psychology

BACKGROUND: For patients with drug-resistant epilepsy, surgery may be effective in controlling their disease. Surgical evaluation may involve localization of the language areas using functional magnetic resonance imaging (fMRI) or Wada testing. We evaluated the accuracy of task-based fMRI versus Wada-based language lateralization in a cohort of our epilepsy patients. METHODS: In a single-center, retrospective analysis, we identified patients with medically intractable epilepsy who participated in presurgical language mapping (n = 35) with fMRI and Wada testing. Demographic variables and imaging metrics were obtained. We calculated the laterality index (LI) from task-evoked fMRI activation maps across language areas during auditory and reading tasks to determine lateralization. Possible scores for LI range from -1 (strongly left-hemisphere dominant) to 1 (strongly right-hemisphere dominant). Concordance between fMRI and Wada was estimated using Cohen's Kappa coefficient. Association between the LI scores from the auditory and reading tasks was tested using Spearman's rank correlation coefficient. RESULTS: = 0.001). CONCLUSION: Our findings suggest that fMRI is generally an accurate, low-risk alternative to Wada testing for language lateralization. However, when fMRI indicates atypical language lateralization (e.g., bilateral dominance), patients may benefit from subsequent Wada testing or intraoperative language mapping.

Extracellular adenosine enhances the ability of PMNs to kill <i>Streptococcus pneumoniae</i> by inhibiting IL-10 production

Journal of Leukocyte Biology · 2020 · 34 citations

• Biology
• Microbiology
• Immunology

mice to pneumococcal pneumonia. CD73/IL-10 did not affect apoptosis, bacterial uptake, and intracellular killing or production of antimicrobial neutrophil elastase and myeloperoxidase. Rather, inhibition of IL-10 production by CD73 was important for optimal reactive oxygen species (ROS) production by PMNs. ROS contributed to PMN antimicrobial function as their removal or detoxification impaired the ability of PMNs to efficiently kill S. pneumoniae. This study demonstrates that CD73 controls PMN antimicrobial phenotype during S. pneumoniae infection.