The triple antibiotic formulation, known as anti-MAP therapy, exhibits unique synergistic antimicrobial activity and should be effective for treatment of Crohn's disease (CD) associated with Mycobacterium avium subspecies paratuberculosis (MAP). The absence of MAP detection in some CD cases may be linked to poor diagnostics or lack of association with the disease. To understand the therapeutic response of some CD patients to anti-MAP therapy in absence of MAP detection, the immunomodulatory potency of anti-MAP therapy and its major ingredients, clarithromycin (CLA) and rifabutin (RIF), in THP-1, Caco-2, and Jurkat T-cells were investigated. Anti-MAP formulation at 2.0 µg/mL decreased MAP viability in macrophages by 18-fold over 72 h. Additionally, M1/M2 macrophage polarization ratio was reduced by 6.7-fold, and expression and protein levels of TNF-a and IL-6 were reduced by 2.9-fold, whereas IL-10 increased by 5.0-fold in these cells. Mechanistically, the effect of anti-MAP formulation on NF-kB activation was dose-dependent and decreased to 13.4% at 2.0 µg/mL. Anti-MAP therapy also reversed the pro-inflammatory response in lipopolysaccharide (LPS)-induced macrophages, which shows that the anti-inflammatory effect of the treatment is not just due to a decrease in MAP viability. Furthermore, this study shows that anti-MAP therapy exhibits anti-cytotoxic effects in Caco-2 monolayers infected with MAP or treated with dextran sodium sulfate (DSS). Anti-MAP therapy decreased T-cell proliferation by up to 4.8-fold following treatment with phytohemagglutinin (PHA) or MAP purified protein derivative (PPD). Overall, the data demonstrate that anti-MAP therapy plays a significant role in modulating and eliciting a protective immune response in macrophages, endothelial cells, and T lymphocytes, even in absence of infection. This may explain the therapeutic response of some CD patients to treatment, even in absence of MAP detection, infection, or total eradication. The study supports anti-MAP therapy as an alternate treatment option for CD, especially in absence of reliable MAP diagnostics.
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Master of Science (M.S.)
College of Medicine
Burnett School of Biomedical Sciences
Length of Campus-only Access
Masters Thesis (Open Access)
Elkamel, Erij, "Anti-MAP Triple Therapy Supports Immunomodulatory Therapeutic Response in Crohn's Disease Through Downregulation of NF-kB Activation in the Absence of MAP Detection" (2021). Electronic Theses and Dissertations, 2020-. 499.