sirolimus and Tuberculosis

Activated phosphoinositide 3-kinase δ syndrome (APDS) is a primary immunodeficiency first described in 2013, which is caused by gain-of-function mutations in PIK3CD or PIK3R1, and characterized by recurrent respiratory tract infections, lymphoproliferation, herpesvirus infection, autoimmunity, and enteropathy. We sought to review the clinical phenotypes, immunological characteristics, treatment, and prognosis of APDS in a large genetically defined Chinese pediatric cohort.. Clinical records, radiology examinations, and laboratory investigations of 40 APDS patients were reviewed. Patients were contacted via phone call to follow up their current situation.. Sinopulmonary infections and lymphoproliferation were the most common complications in this cohort. Three (10.3%) and five (12.5%) patients suffered localized BCG-induced granulomatous inflammation and tuberculosis infection, respectively. Twenty-seven patients (67.5%) were affected by autoimmunity, while malignancy (7.5%) was relatively rare to be seen. Most patients in our cohort took a combined treatment of anti-infection prophylaxis, immunoglobulin replacement, and immunosuppressive therapy such as glucocorticoid or rapamycin administration. Twelve patients underwent hematopoietic stem cell transplantation (HSCT) and had a satisfying prognosis.. Clinical spectrum of APDS is heterogeneous. This cohort's high incidence of localized BCG-induced granulomatous inflammation and tuberculosis indicates Mycobacterial susceptibility in APDS patients. Rapamycin is effective in improving lymphoproliferation and cytopenia. HSCT is an option for those who have severe complications and poor response to other treatments.

Topics: BCG Vaccine; Child; China; Class I Phosphatidylinositol 3-Kinases; Humans; Inflammation; Primary Immunodeficiency Diseases; Sirolimus; Tuberculosis

Tuberculosis (TB) treatment regimens are lengthy, causing non-adherence to treatment. Inadequate treatment can lead to relapse and the development of drug resistance TB. Furthermore, patients often exhibit residual lung damage even after cure, increasing the risk for relapse and development of other chronic respiratory illnesses. Host-directed therapeutics are emerging as an attractive means to augment the success of TB treatment. In this study, we used C3HeB/FeJ mice as an experimental model to investigate the potential role of rapamycin, a mammalian target of rapamycin inhibitor, as an adjunctive therapy candidate during the treatment of Mycobacterium tuberculosis infection with moxifloxacin. We report that administration of rapamycin with or without moxifloxacin reduced infection-induced lung inflammation, and the number and size of caseating necrotic granulomas. Results from this study strengthen the potential use of rapamycin and its analogs as adjunct TB therapy, and importantly underscore the utility of the C3HeB/FeJ mouse model as a preclinical tool for evaluating host-directed therapy candidates for the treatment of TB.

Topics: Animals; B-Lymphocytes; Cell Aggregation; Disease Models, Animal; Female; Lung; Mice; Moxifloxacin; Mycobacterium tuberculosis; Necrosis; Neutrophil Infiltration; Polymethacrylic Acids; Sirolimus; Tuberculosis

Human macrophages play a major role in controlling tuberculosis (TB), but their anti-mycobacterial mechanisms remain unclear among individuals with metabolic alterations like obesity (TB protective) or diabetes (TB risk). To help discern this, we aimed to: i) Evaluate the impact of the host's TB status or their comorbidities on the anti-mycobacterial responses of their monocyte-derived macrophages (MDMs), and ii) determine if the autophagy inducer rapamycin, can enhance these responses. We used MDMs from newly diagnosed TB patients, their close contacts and unexposed controls. The MDMs from TB patients had a reduced capacity to activate T cells (surrogate for antigen presentation) or kill M. tuberculosis (Mtb) when compared to non-TB controls. The MDMs from obese participants had a higher antigen presenting capacity, whereas those from chronic diabetes patients displayed lower Mtb killing. The activation of MDMs with rapamycin led to an enhanced anti-mycobacterial activity irrespective of TB status but was not as effective in patients with diabetes. Further studies are warranted using MDMs from TB patients with or without metabolic comorbidities to: i) elucidate the mechanisms through which host factors affect Mtb responses, and ii) evaluate host directed therapy using autophagy-inducing drugs like rapamycin to enhance macrophage function.

Topics: Adolescent; Adult; Anti-Bacterial Agents; Autophagy; Cross-Sectional Studies; Diabetes Mellitus; Female; Humans; Macrophages; Male; Middle Aged; Mycobacterium tuberculosis; Obesity; Sirolimus; Tuberculosis; Young Adult

In human primary dendritic cells (DC) rapamycin-an autophagy inducer and protein synthesis inhibitor-overcomes the autophagy block induced by

Topics: Autophagy; Cells, Cultured; Cytokines; Dendritic Cells; Gene Expression Profiling; Glycogen Synthase Kinase 3 beta; Humans; Mycobacterium tuberculosis; Primary Cell Culture; Signal Transduction; Sirolimus; TOR Serine-Threonine Kinases; Tuberculosis

To survive and replicate in macrophages Mycobacterium tuberculosis (Mtb) has developed strategies to subvert host defence mechanisms, including autophagy. Autophagy induction has the potential to clear Mtb, but little is known about its effect during controlled tuberculosis and HIV co-infection. Mammalian target of rapamycin complex1 (mTORC1) inhibitors were used to induce autophagy in human macrophages pre-infected with HIV-1BaL and infected with a low dose of Mtb (co-infected), or single Mtb infected (single infected). The controlled Mtb infection was disrupted upon mTOR inhibition resulting in increased Mtb replication in a dose-dependent manner which was more pronounced during co-infection. The increased Mtb replication could be explained by the marked reduction in phagosome acidification upon mTOR inhibition. Autophagy stimulation targeting mTORC1 clearly induced a basal autophagy with flux that was unlinked to the subcellular environment of the Mtb vacuoles, which showed a concurrent suppression in acidification and maturation/flux. Overall our findings indicate that mTOR inhibition during Mtb or HIV/Mtb co-infection interferes with phagosomal maturation, thereby supporting mycobacterial growth during low-dose and controlled infection. Therefore pharmacological induction of autophagy through targeting of the canonical mTORC1-pathway should be handled with caution during controlled tuberculosis, since this could have serious consequences for patients with HIV/Mtb co-infection.

Topics: Autophagy; Coinfection; Gene Expression Regulation; HIV Infections; Host-Pathogen Interactions; Humans; Lysosomes; Macrophages; Mechanistic Target of Rapamycin Complex 1; Microtubule-Associated Proteins; Mycobacterium tuberculosis; Naphthyridines; Phagosomes; Phosphorylation; Sequestosome-1 Protein; Sirolimus; Tuberculosis

Cells in homeostasis metabolize glucose mainly through the tricarboxylic acid cycle and oxidative phosphorylation, while activated cells switch their basal metabolism to aerobic glycolysis. In this study, we examined whether metabolic reprogramming toward aerobic glycolysis is important for the host response to Mycobacterium tuberculosis (Mtb). Through transcriptional and metabolite analysis we show that Mtb induces a switch in host cellular metabolism toward aerobic glycolysis in human peripheral blood mononuclear cells (PBMCs). The metabolic switch is TLR2 dependent but NOD2 independent, and is mediated in part through activation of the AKT-mTOR (mammalian target of rapamycin) pathway. We show that pharmacological inhibition of the AKT/mTOR pathway inhibits cellular responses to Mtb both in vitro in human PBMCs, and in vivo in a model of murine tuberculosis. Our findings reveal a novel regulatory layer of host responses to Mtb that will aid understanding of host susceptibility to Mtb, and which may be exploited for host-directed therapy.

Topics: Animals; Anti-Bacterial Agents; Gene Expression Profiling; Glucose; Glycolysis; Host-Pathogen Interactions; Humans; Leukocytes, Mononuclear; Mice; Mycobacterium tuberculosis; Oxidative Phosphorylation; Proto-Oncogene Proteins c-akt; Signal Transduction; Sirolimus; Toll-Like Receptor 2; TOR Serine-Threonine Kinases; Tuberculosis

To study the effect of Bacilli Calmette-Guerin (BCG) on miRNAs (miR-21, miR-181a, miR-155 and miR-144) in RAW264.7 cells, and with miR-144 as an example, to verify the relationship of miR-144 and Atg4a, an autophagy-related gene, and study the mechanism underlying the regulatory effect of miR-144 on autophagy in the process of Mycobacterium tuberculosis infection.. RAW264.7 cells were treated respectively with starvation (12 hours), 50 ng/mL rapamycin (2 hours) and 10 nmol/L 3-methyl adenine (3-MA, 12 hours), and then stimulated with BCG for 12, 24 or 48 hours. The macrophages were collected for total RNA extraction. The expression levels of miR-21, miR-181a, miR-155 and miR-144 were detected by real-time quantitative PCR (qRT-PCR). Thereafter, we constructed the recombinant plasmids pMIR-Report-Atg4a and pMIR-Report-Atg4a mut. The targeting effect of miR-144 on Atg4a gene was verified by the dual-luciferase reporter assay system, Western blotting and qRT-PCR.. After BCG stimulated RAW264.7 cells, the expression levels of miR-21, miR-155 and miR-144 were up-regulated, and miR-181a expression was down-regulated. The expressions of miR-21, miR-144, miR-155 and miR-181a increased 64 times, 52 times, 14 times and 1 times in the rapamycin group, respectively; the expressions of miR-21, miR-144, miR-155 and miR-181a decreased 1.22 times, 1.05 times, 1.54 times and 12.5 times in the 3-MA group, respectively. The dual-luciferase reporter assay system and Western blotting demonstrated that miR-144 could suppress Atg4a expression by targeting the specific 3'-untranslated region (3'UTR) sequence of Atg4a gene.. miR-144 can directly inhibit the autophagy-related gene Atg4a expression and participate in the regulation of autophagy process in Mycobacterium tuberculosis infection.

Topics: Animals; Autophagy; Base Sequence; BCG Vaccine; Cattle; Cell Line; Cysteine Endopeptidases; Down-Regulation; Humans; Mice; MicroRNAs; Molecular Sequence Data; Mycobacterium bovis; Mycobacterium tuberculosis; Sirolimus; Tuberculosis

This study was designed to investigate the role of the phosphatidyl inositol 3-kinase (PI3K)/AKT/p70(S6K) signaling path on regulation of primary normal human bronchial epithelial cell-derived matrix metalloproteinase (MMP)-1, -3, and -9 expression in tuberculosis (TB). These MMPs are key in pathological extracellular matrix degradation in TB. Normal human bronchial epithelials were stimulated with conditioned medium from monocytes infected with virulent TB (CoMTb) and components of the PI3K/AKT signaling pathway blocked using specific chemical inhibitors and siRNA. MMP gene expression was measured by RT-PCR and secretion by ELISA, luminex, or zymography. Phospho-p70 S6K was detected by Western blot analysis and activity blocked by rapamycin. Chemical blockade of the proximal catalytic PI3K p110 subunit augmented MMP-1 and MMP-9 in a dose-dependent manner (all P<0.001) but suppressed MMP-3 (P<0.01). Targeted siRNA studies identified the p110α isoform as key causing 5-fold increase in TB network-dependent MMP-1 secretion to 4900 ± 1100 pg/ml. Specific inhibition of the AKT node suppressed all 3 MMPs. Phospho-p70(S6K) was identified in the cellular model, and rapamycin, a p70(S6K) inhibitor, inhibited MMP-1 (P<0.001) and MMP-3 (P<0.01) but not MMP-9. Controls were epithelial cells that were unstimulated or exposed to conditioned medium from monocytes not exposed to TB. In summary, blockade of the proximal PI3K catalytic subunit increases MMP-1 and MMP-9, whereas rapamycin decreased both MMP-1 and MMP-3. The regulation of the PI3K path in TB is complex, MMP specific, and a potential immunotherapeutic target in diseases characterized by tissue destruction.

Topics: Cells, Cultured; Humans; Matrix Metalloproteinase 1; Matrix Metalloproteinase 3; Matrix Metalloproteinase 9; Mycobacterium tuberculosis; Phosphatidylinositol 3-Kinase; Ribosomal Protein S6 Kinases, 70-kDa; Signal Transduction; Sirolimus; Tuberculosis

Topics: Antibodies, Monoclonal, Humanized; Antineoplastic Combined Chemotherapy Protocols; Axitinib; Bevacizumab; Carcinoma, Renal Cell; Clinical Trials, Phase III as Topic; Humans; Imidazoles; Indazoles; Kidney Neoplasms; Male; Middle Aged; Randomized Controlled Trials as Topic; Sirolimus; Tuberculosis

Th1 immunity protects against tuberculosis infection in mice and humans. The widely used BCG vaccine primes CD4 and CD8 T cells through signaling mechanisms from dendritic cells and macrophages. The latter express MHC-II and MHC-I molecules through which peptides from BCG vaccine are presented to CD4 and CD8 T cells, respectively. Since BCG sequesters within a phagosome that does not fuse with lysosomes, generation of peptides within antigen-presenting cells infected with BCG occurs with reduced efficiency. We demonstrate that activation of DCs containing BCG vaccine with rapamycin leads to an enhanced ability of DC vaccines to immunize mice against tuberculosis. Coadministration of rapamycin with BCG vaccine also enhanced Th1 immunity. We propose that rapamycin-mediated increase in Th1 responses offers novel models to study mTOR-mediated regulation of immunity.

Topics: Animals; BCG Vaccine; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Dendritic Cells; Humans; Macrophages; Major Histocompatibility Complex; Mice; Mice, Inbred C57BL; Mycobacterium tuberculosis; Phagosomes; Sirolimus; Th1 Cells; TOR Serine-Threonine Kinases; Tuberculosis

Emerging evidence points to an important role of autophagy in the immune response mediated by dendritic cells (DC) against Mycobacterium tuberculosis (Mtb). Since current vaccination based on Bacillus Calmette-Guerin (BCG) is unable to stop the tuberculosis epidemic, a deeper comprehension of the alterations induced by Mtb in DC is essential for setting new vaccine strategies. Here, we compared the capacity of virulent (H37Rv) and avirulent (H37Ra) Mtb strains as well as BCG to modulate autophagy in human primary DC. We found that Mtb H37Rv impairs autophagy at the step of autophagosome-lysosome fusion. In contrast, neither Mtb H37Ra nor BCG strains were able to hamper autophagosome maturation. Both these attenuated strains have a functional inhibition of the 6kD early secreted antigenic target ESAT-6, an effector protein of the ESAT-6 Secretion System-1(ESX-1)/type VII secretion system. Notably, the ability to inhibit autophagy was fully restored in recombinant BCG and Mtb H37Ra strains in which ESAT-6 secretion was re-established by genetic complementation using either the ESX-1 region from Mtb (BCG::ESX-1) or the PhoP gene (Mtb H37Ra::PhoP), a regulator of ESAT-6 secretion. Importantly, the autophagic block induced by Mtb was overcome by rapamycin treatment leading to an increased interleukin-12 expression and, in turn, to an enhanced capacity to expand a Th1-oriented response. Collectively, our study demonstrated that Mtb alters the autophagic machinery through the ESX-1 system, and thereby opens new exciting perspectives to better understand the relationship between Mtb virulence and its ability to escape the DC-mediated immune response.

Topics: Autophagy; Bacterial Secretion Systems; Biomarkers; Cell Differentiation; Cells, Cultured; Dendritic Cells; Humans; Mycobacterium bovis; Mycobacterium tuberculosis; Phagosomes; Sirolimus; Th1 Cells; Tuberculosis

Reactivation of tuberculosis is rare in patients receiving chemotherapy for solid tumours, and poorly documented in patients receiving molecular targeted therapy. We report on a patient with metastatic renal-cell carcinoma treated with temsirolimus, who developed respiratory symptoms and mild fever after 6 weeks of treatment. CT-scan and laboratory tests were consistent with reactivation of tuberculosis. The patient received anti-tuberculosis therapy including rifampicin, a potent CYP3A4/5 inducer. After introduction of rifampicin-based treatment, the patient experienced tumour progression, leaving questionable the potential pharmacokinetic interaction between rifampicin and temsirolimus, a substrate for CYP3A4.

Topics: Aged, 80 and over; Antibiotics, Antitubercular; Antineoplastic Agents; Carcinoma, Renal Cell; Cytochrome P-450 CYP3A; Drug Interactions; Enzyme Induction; Humans; Kidney Neoplasms; Male; Neoplasm Metastasis; Rifampin; Sirolimus; Tuberculosis