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1. Tumor difference between meropenem and vehicle gps
2. No difference in peripheral blood B cell counts or CAR-T counts
Methods to identify:
Decontamination on top of meropenem
Depletion of good bacteria affects CAR-T response in following ways:
1. Loss of important metabolites in serum
2. Loss of co-stimulatory ligands
3. Increase of suppressive ligands
Bacterial add-back
Use 16S qPCR to identify species-level information
Followed by full length sequencing to identify strain level information
Use 1:1 ratios of various bacteria in add-back experiment to validate contribution of specific species versus multiple species
Use non-functional mutant to validate effect of bacteria
Serum metabolomics
Which metabolites should be lost based on literature?
Bacteria supernatant improves CAR-T cell function/phenotype
What are some stimulatory ligands versus ligands that support activation and differentiation of suppressive ligands?
Expansion of bad bacteria in absence of good bacteria drives poor response
No functional issue since peripheral blood B cell recovery on day 21 is same between both groups