Host–pathogen
Do ‘single-cell’ Mtb preps change macrophage results?
Open access · cc by · source: Europe PMC
Gentle sonication made Mtb hyperinflammatory (TLR2-high IL-1β/TNF) yet still replicative in macrophages, while filtering blunted inflammation and attenuated intracellular survival — so prep method can rewrite host–pathogen readouts.
How this study connects
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Study at a glance
- Design
- Animal / in-vitro — In-vitro Mtb single-cell prep (sonication vs 5 μm filter vs spin) infecting mouse BMDMs; RNA-seq, cytokines, CFU, TEM; literature survey of 119 papers
- N
- N=5 · RNA-seq n=5 BMDM infections per condition (so/sp vs 5μmF, MOI 5, 72 h); plus TEM/CFU and 119-paper methods survey
- Population
- Mycobacterium tuberculosis (H37Rv and others) and mouse bone marrow-derived macrophages, including Tlr2−/−
- Outcome
- Envelope integrity, macrophage inflammatory gene expression/cytokines, and intracellular Mtb survival
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Inspect source passages
Do ‘single-cell’ Mtb preps change macrophage results?
These passages occur verbatim in the stored paper text. They support the points listed below; they do not verify every statement in the explanation.
RNA-seq used n=5 per condition at MOI 5, 72 hpi.
BMDMs were uninfected or infected with Mtb prepared by sonication and spin (so/sp) or filtration (5μmF) at an MOI of 5 and analyzed 72 hpi by RNA-seq (n=5 per condition).
Only 39.5% of 119 papers reported single-cell methods.
Of the 119 papers, only 39.5% reported how they generated single cell suspensions.
Key findings
Both sonication and filtering damaged the envelope. Sonicated bacilli drove higher TLR2-dependent transcription and IL-1β/TNF-α but replicated normally; filtered Mtb were hypo-inflammatory and attenuated. PDIM’s apparent anti-inflammatory effect depended on prep method. Only 39.5% of 119 papers reported how they made single-cell suspensions.
Methodology
Compared routine disaggregation (sonication, 5 μm filtering, spinning) for envelope damage (TEM), infected mouse BMDMs (RNA-seq n=5, cytokines, CFU), tested Tlr2−/− BMDMs, and surveyed how 119 papers reported single-cell methods.
Limitations
This is mouse BMDM work in vitro, not a clinical TB trial; other labs’ strains, detergents, or human macrophages may differ, and no single ‘correct’ prep is proven for all questions.
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