Expression of the innate immune receptor LILRB5 on monocytes is associated with mycobacteria exposure.

Antigen presenting cells (APC) are critical components of innate immunity and consequently shape the adaptive response. Leukocyte Ig Like Receptors (LILR) are innate immune receptors predominantly expressed on myeloid cells. LILR can influence the antigen presenting phenotype of monocytic cells to determine the nature of T cell responses in infections including Mycobaterium leprae. We therefore investigated the relevance of LILR in the context of Mycobacterium tuberculosis. Real-time PCR studies indicated that the transcriptional profile of the orphan receptor LILRB5 was significantly up-regulated following exposure to mycobacteria. Furthermore, LILRA1 and LILRB5 were able to trigger signalling through direct engagement of mycobacteria using tranfectant cells incorporating a reporter system. We describe for the first time the expression of this receptor on T cells, and highlight the potential relevance to mycobacterial recognition. Furthermore, we demonstrate that crosslinking of this receptor on T cells increases proliferation of cytotoxic, but not helper, T cells.

LILRA2 activation inhibits dendritic cell differentiation and antigen presentation to T cells.

The differentiation of monocytes into dendritic cells (DC) is a key mechanism by which the innate immune system instructs the adaptive T cell response. In this study, we investigated whether leukocyte Ig-like receptor A2 (LILRA2) regulates DC differentiation by using leprosy as a model. LILRA2 protein expression was increased in the lesions of the progressive, lepromatous form vs the self-limited, tuberculoid form of leprosy. Double immunolabeling revealed LILRA2 expression on CD14+, CD68+ monocytes/macrophages. Activation of LILRA2 on peripheral blood monocytes impaired GM-CSF induced differentiation into immature DC, as evidenced by reduced expression of DC markers (MHC class II, CD1b, CD40, and CD206), but not macrophage markers (CD209 and CD14). Furthermore, LILRA2 activation abrogated Ag presentation to both CD1b- and MHC class II-restricted, Mycobacterium leprae-reactive T cells derived from leprosy patients, while cytokine profiles of LILRA2-activated monocytes demonstrated an increase in TNF-alpha, IL-6, IL-8, IL-12, and IL-10, but little effect on TGF-beta. Therefore, LILRA2 activation, by altering GM-CSF-induced monocyte differentiation into immature DC, provides a mechanism for down-regulating the ability of the innate immune system to activate the adaptive T cell response while promoting an inflammatory response.

The expression of RAGE and EN-RAGE in leprosy.

BACKGROUND: Extracellular newly identified RAGE-binding protein (EN-RAGE) is a ligand of the receptor for advanced glycation endproducts (RAGE) and has been termed S100A12. The ligation of EN-RAGE with RAGE on the endothelium, mononuclear phagocytes and lymphocytes triggers cellular activation with the generation of the key proinflammatory mediators interleukin (IL)-1beta and tumour necrosis factor (TNF)-alpha. OBJECTIVES: The aim of this study was to investigate the presence of RAGE and EN-RAGE, their spatial localization and their coexpression in leprosy lesions. METHODS: Immunohistochemistry and confocal laser scanning microscopy were used to evaluate the expression of RAGE and EN-RAGE in leprosy. By enzyme-linked immunosorbent assay, RAGE and EN-RAGE were detected in the serum. RESULTS: (1) In the multibacillary (MB) and paucibacillary (PB) groups, the level of RAGE production was significantly higher than in patients with atypical mycobacterial infection or sarcoidosis (P < 0.01). In the MB group, the production of RAGE was higher than in the PB group (P < 0.01), and it was higher in patients without the lepra reaction than in patients with the lepra reaction (P < 0.05). (2) In MB, PB and atypical mycobacterial infection, the level of EN-RAGE production was significantly higher than in sarcoidosis (P < 0.01). (3) In the confocal laser scanning microscopic examination, the RAGE and EN-RAGE proteins were detected in lepromatous leprosy. These proteins are spatially colocalized along the cell surface, which is in agreement with their receptor-ligand interaction. (4) A comparable amount of EN-RAGE was detected in the serum of the MB and PB groups. Patients with the reaction showed a higher level of EN-RAGE than patients without the reaction in leprosy. CONCLUSIONS: Our data suggest that in leprosy, RAGE and EN-RAGE may be involved in the proinflammatory process rather than the antimycobacterial activity, especially during the lepra reaction. The blockade of the interaction of RAGE and EN-RAGE at the early stage of the inflammatory process may minimize the inflammatory response and consequent tissue damage or the sequelae of leprosy.

Use of genetic profiling in leprosy to discriminate clinical forms of the disease.

Leprosy presents as a clinical and immunological spectrum of disease. With the use of gene expression profiling, we observed that a distinction in gene expression correlates with and accurately classifies the clinical form of the disease. Genes belonging to the leukocyte immunoglobulin-like receptor (LIR) family were significantly up-regulated in lesions of lepromatous patients suffering from the disseminated form of the infection. In functional studies, LIR-7 suppressed innate host defense mechanisms by shifting monocyte production from interleukin-12 toward interleukin-10 and by blocking antimicrobial activity triggered by Toll-like receptors. Gene expression profiles may be useful in defining clinical forms of disease and providing insights into the regulation of immune responses to pathogens.

Use of genetic profiling in leprosy to discriminate clinical forms of the disease

Leprosy presents as a clinical and immunological spectrum of disease. With the use of gene expression profiling, we observed that a distinction in gene expression correlates with and accurately classifies the clinical form of the disease. Genes belonging to the leukocyte immunoglobulin-like receptor (LIR) family were significantly up-regulated in lesions of lepromatous patients suffering from the disseminated form of the infection. In functional studies, LIR-7 suppressed innate host defense mechanisms by shifting monocyte production from interleukin-12 toward interleukin-10 and by blocking antimicrobial activity triggered by Toll-like receptors. Gene expression profiles may be useful in defining clinical forms of disease and providing insights into the regulation of immune responses to pathogens.

Expression of adhesion molecules in leprosy lesions.

Leprosy presents as a clinical spectrum that is precisely paralleled by a spectrum of immunological reactivity. The disease provides a useful and accessible model, in this case in the skin, in which to study the dynamics of cellular immune responses to an infectious pathogen, including the role of adhesion molecules in those responses. In lesions characterized by strong delayed-type hypersensitivity against Mycobacterium leprae (tuberculoid, reversal reaction, and Mitsuda reaction), the overlying epidermis exhibited pronounced keratinocyte intracellular adhesion molecule 1 (ICAM-1) expression and contained lymphocytes expressing the ICAM-1 ligand, LFA-1. Conversely, in lesions in which delayed-type hypersensitivity was lacking (lepromatous), keratinocyte ICAM-1 expression was low and LFA-1+ lymphocytes were rare. Expression of these adhesion molecules on the cells within the dermal granulomas was equivalent throughout the spectrum of leprosy. The percentage of lymphocytes in these granulomas containing mRNA coding for gamma interferon and tumor necrosis factor alpha, synergistic regulators of ICAM-1 expression, paralleled epidermal ICAM-1 expression. In lesions of erythema nodosum leprosum, a reactional state of lepromatous leprosy thought to be due to immune complex deposition, keratinocyte ICAM-1 expression and gamma interferon mRNA+ cells were both prominent. Antibodies to LFA-1 and ICAM-1 blocked the response of both alpha beta and gamma delta T-cell clones in vitro to mycobacteria. Overall, the expression of adhesion molecules by immunocompetent epidermal cells, as well as the cytokines which regulate such expression, correlates with the outcome of the host response to infection.

Flow cytometric analysis of CD2 modulation on human peripheral blood T lymphocytes by Dharmendra preparation of Mycobacterium leprae.

It has been reported previously that Mycobacterium leprae modulated CD2 on human peripheral blood T lymphocytes and that this modulation was accompanied by a marked reduction in the proliferative response of these cells to mitogens and antigens. In this study, we report that treatment of peripheral blood mononuclear cells from healthy individuals with Dharmendra preparation of M. leprae inhibited their ability to form rosettes with sheep red blood cells. Flow cytometric analysis of Dharmendra lepromin-treated cells showed that, in addition to CD2, CD4 and CD8 were modulated while the surface expression of CD3 was not affected. The specificity of CD2 modulation was confirmed by similar effects of Dharmendra lepromin on thymocytes and lymph node cells from human CD2 transgenic mice. The modulatory effect of Dharmendra lepromin was not observed at lower temperatures. Dharmendra lepromin treatment of activated T cells resulted in reduced binding of monoclonal antibodies to IL-2R and D66 epitope of CD2. The modulatory effects were not observed with Dharmendra preparation of BCG or other preparations of M. leprae. Our results indicate that certain M. leprae factor(s) specifically modulate(s) CD2, CD4, CD8 and IL-2R but not CD3 on T lymphocytes. The suppressive effect of Dharmendra lepromin on the T-cell proliferative response reported earlier may be explained by its modulatory effect on a number of T-cell surface molecules.

Quantitation of the soluble E-receptor of human T lymphocytes by rocket electrophoresis in the serum of patients with lepromatous and tuberculoid leprosy.

Human T lymphocytes carry a membrane receptor for sheep erythrocytes (E) which is responsible for the well-known phenomenon of E-rosette formation. This receptor has been related to CD2 molecules; it is present in a soluble form (Rs) in normal serum and may play an immunoregulatory role. In this study we quantitated soluble E-receptor in serum samples of 43 normal controls, 32 patients with tuberculoid leprosy and 53 with lepromatous leprosy, using rocket electrophoresis and an anti E receptor serum (anti-Rs) obtained from an adult sheep immunized with autologous E treated with Rs. In the 3 groups studied, the rocket means were respectively 5.0, 7.5 and 10.9 mm (p less than 0.001). We found abnormally high levels of Rs in the serum of various diseases associated with a depression of cell-mediated immunity. The increase of Rs levels in the serum may be one of the mechanisms responsible for the depression of cellular immunity in leprosy.

Quantitation of the soluble E-receptor of human T lymphocytes by rocket electrophoresis in the serum of patients with lepromatous and tuberculoid leprosy

Los linfocitos T humanos poseen, a nivel de membrana, receptores para eritrocitos de carnero (E) que les permite la unión espontánea con estas células, llevando al fenómeno de formación de rosetas. Este receptor fue relacionado con la molécula CD2. Se produjeron varios anticuerpos monoclonales contra la estructura CD2; la mayoría de ellos inhibe alguna de las funciones de los linfocitos T (blasogénesis, formación de rosetas y reacción en cultivo mixto de linfocitos). Es posible obtener este receptor de membrana en forma soluble (Rs), en el sobrenadante de células T incubadas por una hora a 4-C y en el suero de individuos normales. En nuestro laboratorio fueron estandarizados varios métodos para detección y ccuantificacióne de Rs, a partir de un antisuero (anti-Rs) obtenido al inmunizar un carnero con eritrocitos autólogos tratados con Rs (ERs). Este antisuero inhibe la formación de rosetas, es citotóxico para células T, aglutina complejos ERs e identifica linfocitos T por inmunofluorescencia. El método más práctico para cuantificacióne de Rs es el suero de pacientes con patologías asociadas a depresión de la respuesta inmune mediada por células (anemia, lepra, neoplasias, leucemias, linfomas y aplasia medular). Numerosos trabajos han demostrado que los pacientes portadores de lepra presentan alteraciones inmunológicas. Se sabe que los pacientes con lepra lepromatosa presentan deficiencia primaria específica de células T contra antígenos del M. leprae y deficiencia secundaria, inespecífica, de la respuesta inmune celular. En este trabajo, cuantificamos Rs por inmunoelectrdifusión en el suero de 43 individuos normales, 32 pacientes con lepra tuberculoide y 53 con lepra lepromatosa. El promedio de los picos obtenidos fue de 5,0, 7,5 y 10,9 mm., respectivamente. Este aumento fue estadísticamente significativo (Kruskal-Wallis p<0,001), entre los 2 grupos de pacientes y comparados con el grupo control. El aumento de Rs en el suero de los pacientes con lepra puede ser uno de los mecanismos responsables de la depresión de la respuesta inmune celular encontrada en esta enfermedad

Quantitation of the soluble E-receptor of human T lymphocytes by rocket electrophoresis in the serum of patients with lepromatous and tuberculoid leprosy

Los linfocitos T humanos poseen, a nivel de membrana, receptores para eritrocitos de carnero (E) que les permite la unión espontánea con estas células, llevando al fenómeno de formación de rosetas. Este receptor fue relacionado con la molécula CD2. Se produjeron varios anticuerpos monoclonales contra la estructura CD2; la mayoría de ellos inhibe alguna de las funciones de los linfocitos T (blasogénesis, formación de rosetas y reacción en cultivo mixto de linfocitos). Es posible obtener este receptor de membrana en forma soluble (Rs), en el sobrenadante de células T incubadas por una hora a 4-C y en el suero de individuos normales. En nuestro laboratorio fueron estandarizados varios métodos para detección y ccuantificacióne de Rs, a partir de un antisuero (anti-Rs) obtenido al inmunizar un carnero con eritrocitos autólogos tratados con Rs (ERs). Este antisuero inhibe la formación de rosetas, es citotóxico para células T, aglutina complejos ERs e identifica linfocitos T por inmunofluorescencia. El método más práctico para cuantificacióne de Rs es el suero de pacientes con patologías asociadas a depresión de la respuesta inmune mediada por células (anemia, lepra, neoplasias, leucemias, linfomas y aplasia medular). Numerosos trabajos han demostrado que los pacientes portadores de lepra presentan alteraciones inmunológicas. Se sabe que los pacientes con lepra lepromatosa presentan deficiencia primaria específica de células T contra antígenos del M. leprae y deficiencia secundaria, inespecífica, de la respuesta inmune celular. En este trabajo, cuantificamos Rs por inmunoelectrdifusión en el suero de 43 individuos normales, 32 pacientes con lepra tuberculoide y 53 con lepra lepromatosa. El promedio de los picos obtenidos fue de 5,0, 7,5 y 10,9 mm., respectivamente. Este aumento fue estadísticamente significativo (Kruskal-Wallis p<0,001), entre los 2 grupos de pacientes y comparados con el grupo control. El aumento de Rs en el suero de los pacientes con lepra puede ser uno de los mecanismos responsables de la depresión de la respuesta inmune celular encontrada en esta enfermedad (AU)

Distribution of lymphocytes with interleukin-2 receptors (TAC antigens) in reactive lymphoproliferative processes, Hodgkin's disease, and non-Hodgkin's lymphomas. An immunohistologic study of 300 cases.

The authors investigated the distribution of interleukin-2 receptors (TAC antigen) in the lymph nodes of 300 patients with lymphoproliferative disorders. They used fresh-frozen sections to evaluate a possible correlation between the immunophenotype of specific lymphoid disorders and the presence or absence of TAC expression and to determine whether the TAC positivity of lymphoid cells contributes to the characterization of lymphoproliferative processes. All of the cases had previously been studied with a large screening panel of monoclonal antibodies and polyclonal antisera. Among 85 patients with a variety of benign reactive processes, the lymph nodes from 47 contained TAC-bearing lymphocytes in various patterns of distribution. Of 41 patients with Hodgkin's disease, 37 had TAC-bearing lymphocytes. Of 26 B-cell, well-differentiated lymphocytic lymphomas (WDL), 14 were diffusely TAC-positive and one had TAC-bearing cells in random distribution. Six cases of intermediate lymphocytic lymphoma were also studied, and three showed randomly distributed TAC-bearing lymphocytes. Of 19 patients with follicular or follicular and diffuse, poorly differentiated lymphocytic (PDL) lymphoma, 14 were TAC-positive. All 3 diffuse PDL lymphomas studied were TAC-negative. Among 23 cases of B-cell and 5 cases of T-cell mixed cell lymphoma, 15 and three, respectively, had TAC-positive lymphocytes. Of 39 large cell lymphomas (B-cell, 33; T-cell, 6), 14 were TAC-positive. All 13 cases of hairy cell leukemia were diffusely positive. Of 23 T-lymphoblastic lymphomas, only 1 showed positive TAC reactivity, which was focal. Of 5 cases of cutaneous T-cell lymphoma, 2 had TAC-bearing lymphocytes. Our study indicates that the TAC antigen is not lineage-specific, and that it may be expressed by lymphoid cells regardless of their phenotype.

Defective cell-mediated immunity in leprosy: failure of T cells from lepromatous leprosy patients to respond to Mycobacterium leprae is associated with defective expression of interleukin 2 receptors and is not reconstituted by interleukin 2.

Patients with lepromatous leprosy (LL) but not borderline tuberculoid leprosy (BT) have defective cell-mediated immune responses to Mycobacterium leprae, despite normal responses to other stimuli, as judged by in vivo skin testing and in vitro lymphocyte transformation. To investigate the basis of the immune defect in LL patients, we studied the ability of patient mononuclear leukocytes to produce interleukin 1 (IL 1) and interleukin 2 (IL 2) upon stimulation with M. leprae, and determined the ability of exogenous IL 1 and IL 2 to reconstitute the LL patient response to this antigen in vitro. Equal numbers of adherent non-T cells from LL and BT patients produced similar amounts of IL 1 upon challenge with M. leprae, and addition of IL 1 to the culture medium failed to reconstitute the response of lymphocytes from LL patients to M. leprae. On the other hand, T cells of LL patients failed to express receptors for IL 2 or to produce IL 2 in response to M. leprae, whereas similarly treated T cells of BT patients both expressed IL 2 receptors and produced IL 2. Finally, recombinant human IL 2 purified to homogeneity as well as crude supernatants of mitogen-activated lymphocytes failed to reconstitute the response of LL patients to M. leprae. These results suggest that T cells of LL patients fail to respond to M. leprae despite an ability to produce IL 1 and that their failure to express receptors for IL 2 may explain both defective proliferation and the failure of exogenous IL 2 to reconstitute the response.