Objective--To evaluate the role of the nuclear factor-κB (NF-κB) in the response of bovine monocytes to exposure to Mycobacterium avium subsp paratubercuIosis (MAP). Sample Population--Monocytes from healthy adult Holstein cows that were known to be negative for MAP infection. Procedures--Monocytes were incubated with MAP organisms with or without a specific inhibitor of the NF-κB pathway (pyrrolidine dithiocarbamate), and activation of the NF-κB pathway was detected by use of an electrophorectic mobility shift assay. The capacities of monocytes to express tumor necrosis factor (TNF)-α, interleukin (IL)-10, and IL-12; to acidify phagosomes; to phagocytize and kill MAP organisms; and to undergo apoptosis were evaluated. Results--Addition of MAP organisms to monocytes activated the NF-κB pathway as indicated by increased NF-κB-DNA binding. Addition of pyrrolidine dithiocarbamate prevented nuclear translocation of NF-κB, decreased expression of TNF-α and IL-10, and increased IL-12 expression. Treatment of MAP-exposed monocytes with pyrrolidine dithiocarbamate increased the rate of apoptosis but failed to alter phagosome acidification, organism uptake, or organism killing by those cells. Conclusions and Clinical Relevance--Results indicated that NF-κB rapidly translocated to the nucleus after exposure of bovine monocytes to MAP organisms. These data suggest that NF-κB is involved in initiation of inflammatory cytokine transcription and inhibition of apoptosis but that it is not directly involved in phagosome acidification or organism killing. [ABSTRACT FROM AUTHOR]