Single-cell profiling reveals that dynamic lung immune responses distinguish protection from susceptibility to tuberculosis.

The mechanisms that underlie protective immunity to Mycobacterium tuberculosis (Mtb) remain incompletely defined. To identify immune correlates associated with protection, we performed single-cell RNA sequencing of lung immune cells after aerosol Mtb infection of naïve mice and mice with contained Mtb infection (CoMtb), a model of naturally acquired resistance, across multiple time points, mouse strains, and Mtb strains. Protection was associated with distinct temporal patterns of immune activation, cell recruitment, and resolution. Early after challenge, CoMtb mice exhibited rapid, transient recruitment and activation of CD4 ⁺ T cells, macrophages, NK, and NKT cells, accompanied by short-lived bursts of type I and II interferon signaling, increased oxidative phosphorylation, and enhanced chemokine-mediated cell-cell communication. In contrast, primary infection elicited delayed but sustained interferon and neutrophil responses and higher bacterial burdens. These data indicate that protection involves dynamically coordinated immune pathways rather than the magnitude of any single response. Transcriptional features of CoMtb overlapped with those observed in nonhuman primates following intravenous BCG vaccination, including enrichment of activated tissue-resident CD4 ⁺ T cells and innate effector populations. Together, these findings support a model in which effective immunity to Mtb depends on the timing and coordination of immune activation, providing a framework for vaccine strategies that reproduce protective lung immune dynamics.