Evidence for a More Disrupted Immune-Endocrine Relation and Cortisol Immunologic Influences in the Context of Tuberculosis and Type 2 Diabetes Comorbidity.

Pulmonary tuberculosis (PTB), caused by Mycobacterium tuberculosis (Mtb), is a major health problem worldwide, further aggravated by the convergence of type 2 diabetes mellitus (DM) which constitutes an important risk factor for TB development. The worse scenario of patients with PTB and DM may be partly related to a more unbalanced defensive response. As such, newly diagnosed PTB patients with DM (TB+DM, n = 11) or not (TB, n = 21), as well as DM (n = 18) patients and pair matched controls (Co, n = 22), were investigated for the circulating immuno-endocrine-metabolic profile (ELISA), along with studies in peripheral blood mononuclear cells (PBMC) analyzing transcript expression (RT-qPCR) of mediators involved in glucocorticoid functionality. Given the hyperglycemic/hypercortisolemic scenario of TB+DM patients, PBMC were also exposed to stress-related cortisol concentrations (0.1 and 1 µM) and supraphysiologic glucose doses (10, 20, and 40 mM) and assessed for the specific response against Mtb stimulation (lymphoproliferation, -thymidine incorporation-, and cytokine production -bead-cytometry). All TB patients displayed increased plasma amounts of cortisol, growth hormone -hGH-, and proinflammatory mediators. In turn, TB+DM showed even higher levels of interferon gamma -IFN-γ- and hGH (vs. TB), or IL-6, C reactive protein, cortisol and hGH (vs. DM). Both DM groups had equally augmented values of IL-10. All TB patients showed decreased dehydroepiandrosterone- sulfate concentrations, even more in TB+DM cases. Leptin was also decreased in both TB cases, particularly in the TB group, revealing a lower body mass index, as well. Unlike PBMC from TB cases showing a decreased relationship between the glucocorticoids receptor (GR) isoforms (GRα/GRß; functional isoform/negative isoform), cells from TB+DM patients had no changes in this regard, along with an increased expression of 11-beta hydroxysteroid dehydrogenase type-1, the enzyme facilitating intracellular cortisone to cortisol conversion. TB+DM patients also showed an increased Mtb antigen-driven lymphoproliferation. Compared to TB, DM and HCo counterparts, PBMC from TB+DM patients had a biased Th1 response to Mtb stimulation (increased IL-2 and IFN-γ production), even when exposed to inhibitory cortisol doses. TB+DM patients show a more unbalanced immuno-endocrine relationship, respect the non-diabetic counterparts, with a relative deficiency of cortisol immunomodulatory influences, despite their more favorable microenvironment for cortisol-mediated immune effects.

Tuberculosis, the Disrupted Immune-Endocrine Response and the Potential Thymic Repercussion As a Contributing Factor to Disease Physiopathology.

Upon the pathogen encounter, the host seeks to ensure an adequate inflammatory reaction to combat infection but at the same time tries to prevent collateral damage, through several regulatory mechanisms, like an endocrine response involving the production of adrenal steroid hormones. Our studies show that active tuberculosis (TB) patients present an immune-endocrine imbalance characterized by an impaired cellular immunity together with increased plasma levels of cortisol, pro-inflammatory cytokines, and decreased amounts of dehydroepiandrosterone. Studies in patients undergoing specific treatment revealed that cortisol levels remained increased even after several months of initiating therapy. In addition to the well-known metabolic and immunological effects, glucocorticoids are involved in thymic cortical depletion with immature thymocytes being quite sensitive to such an effect. The thymus is a central lymphoid organ supporting thymocyte T-cell development, i.e., lineage commitment, selection events and thymic emigration. While thymic TB is an infrequent manifestation of the disease, several pieces of experimental and clinical evidence point out that the thymus can be infected by mycobacteria. Beyond this, the thymic microenvironment during TB may be also altered because of the immune-hormonal alterations. The thymus may be then an additional target of organ involvement further contributing to a deficient control of infection and disease immunopathology.

miR-30c is specifically repressed in patients with active pulmonary tuberculosis.

Tuberculous pleurisy (PLTB) is a common form of extrapulmonary tuberculosis. It often resolves without chemotherapy being hence considered a rather benign manifestation of the disease. Patients with PLTB mount an effective anti-mycobacterial response, unlike those with active pulmonary TB (pTB) that were shown to present an imbalance in plasma immune and endocrine mediators. In this work, we explored whether expression of the active isoform of the glucocorticoid receptor (hGRα) in the context of the inflammatory-anti-inflammatory responses of TB patients may be associated to microRNA levels. As expected, the inflammatory response triggered in patients coexists with increased circulating cortisol and altered hGRα levels in the peripheral blood mononuclear cells. However, while hGRα expression is significantly downregulated in PLTB, its levels in pTB patients are higher within the control values. These results point out to the existence of an additional mechanism tending to preserve hGRα levels probably to deal with the chronic inflammation observed in pTB. In this regard, we found that miR-30c is strongly downregulated in mononuclear cells of pTB patients compared to PLTB cases, showing an expression profile opposite to that seen with hGRα. Interestingly, low levels of miR-30c are specific for this active form of TB, as its expression is not altered in mononuclear cells from either healthy controls or patients with tuberculous or non-tuberculous pleurisy. Moreover, miR-30c and hGRα also showed an inverse expression pattern in M. tuberculosis-stimulated THP-1 macrophage cultures. In sum, our studies identify miR-30c as a specific correlate of pulmonary manifestations of TB, potentially involved in the altered glucocorticoid sensitivity observed in these patients.

Altered microRNA expression levels in mononuclear cells of patients with pulmonary and pleural tuberculosis and their relation with components of the immune response.

Different lines of evidence demonstrate that microRNAs (miRNAs) play an important role in host-pathogen interactions. In this study we investigated the expression patterns of several miRNAs, most of them involved in regulating inflammatory responses, in patients with tuberculosis (TB). In order to understand the events occurring at the site of infection, we employed mononuclear cells obtained from both peripheral blood (PBMC) and pleural fluids (PFMC) of patients. Interestingly, we found that the miRNA signature of each compartment is different, with a strong down-regulation in PFMCs of miR-223, miR-144* and miR-421. In addition, we observed that miR-146a expression is also down-regulated in tuberculosis patients, both in PBMCs and PFMCs while miR-424 levels are elevated only in the peripheral compartments. We also showed that systemic expression of these miRNAs changes upon specific treatment and is associated with IL-6 levels, a cytokine playing a substantial role in TB immunopathology. Present results contribute to a better knowledge of the host responses in TB pathogenesis, pointing out the role of miRNAs in this disease.

Influencia de los macrofagos intratumorales en el crecimiento de un sarcoma de rata / Influence of intratumoral macrophages on the growth of a rat sarcoma

La eliminación de las células fagocíticas activas (CFA) de las suspensiones de um sarcoma de rata (S-E 100), provoca um menor desarrollo del mismo en ratas de línea "m", probablemente porque esas células que infiltran el tumor poseen funciones inhibitorias de la respuesta inmune antitumoral. En este trabajo se investiga si el efecto atribuido a las CFA es general, o depende del tipo de interación que se genera entre los macrófagos (MÝ) que infiltran el tumor donante y el huesped receptor. El S-E 100 se inoculó en ratas de línea "m" (S-E 100) y de línea "c" (S-E 100, c); las CFA se eliminaron con hierro carbonílico (FeC), inoculando las células sobrenadantes (S.FeC - m) y (S.FeC-c) y las suspensiones celulares testigos (S.Te-m) y (S.Te - c). Todos los inóculos fueron de 1 x 10**6 células vía s.c., enreceptores adultos "m" y "c". La eliminación de las CFA indujo un menor desarrollo del tumor en receptores "m", sólo si el inóculo provenía de S-E 100,m. En el receptor "c" no se observó ninguna variación en el crecimiento del tumor, tanto si el inóculo prevenía de S-E 100, como de S-E 100,c.Se postula que el efecto inhibidor de la respuesta inmune ejercio por los MÝ que infiltran el S-E 100 no es un efecto general, sino particular de una línea y sólo obtiene si los MÝ intratumorales son singénicos con el receptor

Influencia de los macrofagos intratumorales en el crecimiento de un sarcoma de rata / Influence of intratumoral macrophages on the growth of a rat sarcoma

La eliminación de las células fagocíticas activas (CFA) de las suspensiones de um sarcoma de rata (S-E 100), provoca um menor desarrollo del mismo en ratas de línea "m", probablemente porque esas células que infiltran el tumor poseen funciones inhibitorias de la respuesta inmune antitumoral. En este trabajo se investiga si el efecto atribuido a las CFA es general, o depende del tipo de interación que se genera entre los macrófagos (MY) que infiltran el tumor donante y el huesped receptor. El S-E 100 se inoculó en ratas de línea "m" (S-E 100) y de línea "c" (S-E 100, c); las CFA se eliminaron con hierro carbonílico (FeC), inoculando las células sobrenadantes (S.FeC - m) y (S.FeC-c) y las suspensiones celulares testigos (S.Te-m) y (S.Te - c). Todos los inóculos fueron de 1 x 10**6 células vía s.c., enreceptores adultos "m" y "c". La eliminación de las CFA indujo un menor desarrollo del tumor en receptores "m", sólo si el inóculo provenía de S-E 100,m. En el receptor "c" no se observó ninguna variación en el crecimiento del tumor, tanto si el inóculo prevenía de S-E 100, como de S-E 100,c.Se postula que el efecto inhibidor de la respuesta inmune ejercio por los MY que infiltran el S-E 100 no es un efecto general, sino particular de una línea y sólo obtiene si los MY intratumorales son singénicos con el receptor (AU)