Macrophages are important innate immune cells that respond to microbial insults. In response to multi-bacterial infection, the macrophage activation state may change upon exposure to nascent mediators, which results in different bacterial killing mechanism(s). In this study, we utilized two respiratory bacterial pathogens, Mycobacterium bovis (Bacillus Calmette Guẻrin, BCG) and Francisella tularensis live vaccine strain (LVS) with different phagocyte evasion mechanisms, as model microbes to assess the influence of initial bacterial infection on the macrophage response to secondary infection. Non-activated (M0) macrophages or activated M2-polarized cells (J774 cells transfected with the mouse IL-4 gene) were first infected with BCG for 24–48 h, subsequently challenged with LVS, and the results of inhibition of LVS replication in the macrophages was assessed. BCG infection in M0 macrophages activated TLR2-MyD88 and Mincle-CARD9 signaling pathways, stimulating nitric oxide (NO) production and enhanced killing of LVS. BCG infection had little effect on LVS escape from phagosomes into the cytosol in M0 macrophages. In contrast, M2-polarized macrophages exhibited enhanced endosomal acidification, as well as inhibiting LVS replication. Pre-infection with BCG did not induce NO production and thus did not further reduce LVS replication. This study provides a model for studies of the complexity of macrophage activation in response to multi-bacterial infection.
Digital Object Identifier (DOI)
The authors thank Dr Chinnaswamy Jagannath at the University of Texas Health Science Center at Houston who kindly provided the GFP-expressing BCG under support from his National Institutes of Health (NIH) grant (AI-78420). Image flow analyses were conducted in the University of Texas at San Antonio Immune Defense Core (supported by Research Centers in Minority Institutions, NIH grant G12MD007591).
The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: funding from the Army Research Office of the Department of Defense under contract no. W911NF-11-1-0136, and by the Jane and Roland Blumberg Professorship in Biology, to BPA.
Trivedi, Nikita H.; Yu, Jieh-Juen; Hung, Chiung-Yu; Doelger, Richard P.; Navara, Christopher S.; Armitige, Lisa Y.; Seshu, Janakiram; Sinai, Anthony P.; Chambers, James P.; Guentzel, M. Neal; and Arulanandam, Bernard P., "Microbial Co-Infection Alters Macrophage Polarization, Phagosomal Escape, and Microbial Killing" (2018). Microbiology, Immunology, and Molecular Genetics Faculty Publications. 109.