Clinical isolates of Mycobacterium avium complex reveal an M. intracellulare-associated IL-17/neutrophilic pulmonary immune program in a murine disease model

Mycobacterium avium complex (MAC) is the leading cause of nontuberculous mycobacterial pulmonary disease (NTM-PD) and mainly comprises M. avium (MAV) and M. intracellulare (MI). Host-pathogen interactions may contribute to the heterogeneous clinical course of MAC pulmonary disease (MAC-PD); however, species- or isolate-associated differences in virulence and host immune responses induced by MAC strains remain poorly understood. Here, we established a panel of MAC clinical isolates exhibiting persistent pulmonary infection in mice and performed transcriptomic analyses to evaluate pulmonary immune responses. Although MAC infections induced broadly shared inflammatory responses, MI infection elicited a robust IL-17/neutrophilic inflammatory signature, whereas MAV infection showed relative enrichment of IFN-{gamma}/cytotoxicity-associated responses, indicating differences in the balance of their immune gene expression programs. The MI-associated IL-17/neutrophilic program remained evident at the isolate level and after adjustment for pulmonary growth phenotype. RT-qPCR and flow cytometric analyses using the representative isolate pair FKJ-1 (MI) and FKJ-8 (MAV) further supported the differential induction of these immune signatures in infected lungs. To define the cellular basis of these immune programs, we performed single-cell RNA sequencing on lungs infected with FKJ-1 and FKJ-8. Infection with the representative MI isolate was associated with increased Il17a-expressing CD4+ T cells and {gamma}{delta} T cells, neutrophilic inflammation, and the expansion of inflammatory macrophages. Taken together, these findings demonstrate that within a selected panel of persistent MAC clinical isolates, MI infection is associated with a robust IL-17/neutrophilic pulmonary immune program, providing a preclinical framework for dissecting species- and isolate-associated host-pathogen interactions in MAC-PD.