Long-term omega-3 fatty acids modulate immune responses more effectively than ibuprofen in Mtb-infected mice: An in silico functional analysis approach

Tuberculosis (TB) remains a major global public health threat, necessitating alternative and novel treatments to modulate inflammation and shorten drug treatment periods. The potential of omega-3 long-chain polyunsaturated fatty acids (EPA/DHA) and ibuprofen as adjunct therapies in TB treatment was demonstrated recently in Mycobacterium TB (Mtb) C3HeB/FeJ mice, but the underlying molecular mechanisms of the immune response pathways remain largely unknown. Here, we used in silico predictive functional network modelling to predict underlying molecular relationships and functional network interactions associated with inflammatory and immune response pathways. We focused on Th1, Th2 and Th17 cell pathways in the lung tissue when administering adjunct EPA/DHA or ibuprofen to Mtb-infected C3HeB/FeJ mice. Mice (n = 36) initially received Rifafour® (rifampicin, isoniazid, pyrazinamide, and ethambutol) for three days before transitioning to rifampicin and isoniazid alone for the remainder (11 days) of the experiment, with or without EPA/DHA (1.6:1) or ibuprofen (0.05 g/L) supplementation. Standard TB drug treatment alone downregulated Th1 cell responses, shifting toward a Th2-skewed immune profile, which may impact long-term host immune competence. Adjunct ibuprofen resulted in excessive inflammation as indicated by increased pro-inflammatory cytokines and neutrophil recruitment (via Th17 cell activity), which was associated with higher lung bacterial loads and reduced alveolar space compared to TB drugs alone or the adjunct EPA/DHA group. In contrast, supplementing EPA/DHA maintained a Th1/Th2 immune balance and reduced excessive Th17 activity, yielding lower bacterial loads. These findings suggest that EPA/DHA supplementation holds promise as a safe and more effective host-directed therapy, adjunct to TB drugs, by enhancing immune balance and mitigating excessive lung damage. Further investigation into the clinical applicability of EPA/DHA as an adjunct in TB treatment is warranted.