The mechanisms that contribute to protection from tumor recurrence after M.Bovis BCG are not fully elucidated. We aim to evaluate the role of Antibody Dependent Cell Cytotoxicity (ADCC) in BCG therapy for NMIBC.METHODS: Formalin-fixed and paraffin-embedded (FFPE) tissue sections before and after BCG therapy were used for bulk targeted gene expression analysis. Differential gene expression analysis was performed on targeted RNAseq data. Gene Set Enrichment Analysis (GSEA) was performed using the "Hallmark Gene Sets" from the Broad Intitute's Molecular Signatures Database (MSigDB) and customized assembled gene sets. Enzyme linked immunosorbent assay (ELISA) and OLink Proteomics Ò inflammation panel were used to profile IgG1/3, and Interferon Gamma (IFN-g), respectively on prospectively collected plasma from BCG-treated patients (N[14) at specific timepoints throughout the treatment.RESULTS: B cell markers and IFN-g signatures were the two most enriched pathways after BCG therapy in the GSEA (Fig. 1A). We also observed significantly increased gene expression of IgG1 and IgG3 after BCG therapy (Fig. 1B), which may induce ADCC reactivity via FcgRIIIA signaling. In non-recurrent cases, we observed an increase in plasma IgG1/3 concentration that preceded IFN-g hyperactivity after 1st and 3rd BCG doses (Fig. 2). In contrast, patients that recur after BCG therapy had higher levels of plasma IFN-g at baseline, which remained elevated throughout the induction phase unrelated to IgG1/3 concentrations (Fig. 2). While the median recurrence time was 3.9 months, the median follow-up for nonrecurrent patients was 36.7 months. CONCLUSIONS: We observed increased expression of B cellrelated genes and an IFN-g signature after BCG therapy. Moreover, we found that response to BCG may include activation of Immune cells through IgG1/IgG3-mediated ADCC reactivity. Future studies in larger patient cohorts should measure IFN-g during BCG therapy and consider a role for ADCC in mediating protection from tumor recurrence BCG therapy.
The microbiome of the urinary tract has a much greater association with USD than the gut microbiota, concomitant with reduced functional microbial networks. Furthermore, a diverse but consistent microbiome is often found in kidney stones regardless of composition. These data suggest that a renal microbiome (RM) both exists and is modifiable. Therefore, we aimed to determine the composition and localization of the RM and how it is impacted by antibiotic use.METHODS: We conducted a diet trial with Webster mice fed with a 1.5% oxalate diet housed in metabolic cages. Four groups and four control arms were maintained on cefazolin for 8 or 19 days, with or without 14-days recovery. Urine was collected in 3-days intervals to quantify changes to the urinary microbiome with plate cultures and metagenomic bacterial inventories. Additionally, kidneys were collected for 16s rRNA sequencing after host DNA removal, or for direct bacteria visualization in bisected kidneys with fluorescence in situ hybridization (FISH) on a confocal laser microscope using one of three FISH probes that selected for all bacteria, Lactobacilli (Lact), or Enterobacteraceae (Ent). To evaluate their specificity, all probes were first tested against pure cultures of Lactobacillus sp. and Enterobacter sp.RESULTS: Significant differences in the urinary tract microbiome were observed with long-term antibiotics relative to Lact and Ent abundance with culture based approaches (p[0.02, p[0.04, respectively). Bacterial DNA was present in all the kidney samples processed. FISH probes exhibited their expected specificity for Lact or Ent, and in kidney tissue we identified the presence of Ent (Figure 1) and Lact. Bacteria were primarily located in the renal medulla in the periphery of the loop of Henle.CONCLUSIONS: We were able to demonstrate the presence of bacteria in kidney tissue, and determine their composition and localization, through both bacterial DNA analysis and FISH staining. This has implications for the formation of kidney stones particularly because specific bacteria from the Ent family are commonly found in all types of kidney stones and in the urinary tract of stone formers while Lact in the urinary tract are anti-associated with USD.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.