Differential Th1/Th2 balance in peripheral blood lymphocytes from patients suffering from flea bite-induced papular urticaria

Q4Q2Artículo original7-10Background: The Th1/Th2 balance has not been characterized in patients suffering from flea bite-induced papular urticaria (FBPU). Our aim was to improve understanding of the immunopathogenesis of CD4+ and CD8+ T-cells in humans suffering from flea bite-induced papular urticaria. Methods: Peripheral blood mononuclear cells were obtained from 18 pediatric patients and 10 age-matched healthy controls. Cellular phenotypes, intracellular production of interferon gamma (IFN) and interleukin-4 (IL-4) in T-cells stimulated with polyclonal stimuli was determined by fl ow cytometry following short-term in vitro stimulation. Results: The results revealed lower frequencies of IFN-secreting (p = 0.02) and higher frequencies of IL-4-secreting (p = 0.03) CD4+ T-cells in patient lymphocyte cultures compared to healthy control cultures in the presence of polyclonal stimuli. This is the first description of differential cytokine patterns in papular urticaria patients. Conclusion: Patients suffering from papular urticaria have an atopic status compared to healthy children

Specific pattern of flea antigen recognition by IgG subclass and IgE during the progression of papular urticaria caused by flea bite

Q4Q2Artículo original197-202Background: Papular urticaria caused by flea bite presents clinical symptoms of a hypersensitivity reaction accompanied by skin lesions. However, the pattern of recognition by different antibody isotypes during the progression of the disease is unknown. This study evaluated variations in immunoglobulin E and immunoglobulin G subclass antibody responses to flea antigens during the progression of papular urticaria caused by flea bite Methods: Twenty-five patients clinically diagnosed with papular urticaria due to flea bite were included. Ten healthy children were included as controls. Recognition of antigens from complete flea body extract by patients and healthy controls was determined using immunoblot assays. Results: The results revealed that patients with 2–5 years of papular urticaria evidenced more IgE bands than those with shorter or longer durations of symptoms. In contrast, healthy children showed a predominance of immunoglobulin G1 and immunoglobulin G3. The majority of the recognised antigens were low molecular weight proteins (o90 kDa). Proteins with molecular weights between 16–20, 21–25, and 31–35 kDa showed different patterns of recognition between patients and healthy children. Conclusion: The predominant specific antibody isotypes vary according to the time elapsed since the onset of symptoms in papular urticaria caused by flea bite

GENERACIÓN in vitro DE CÉLULAS DENDRÍTICAS HUMANAS DE ORIGEN MIELOIDE

Dendritic cell activity is an important step to elicit cellular immunity. These cells are the most potent antigen presenting cells and the main activating factor for naive T cells. There are several phenotypes of dendritic cells originated from different cell precursors. The aim of this study was to assess a model to obtain human myeloid dendritic cells from peripheral blood CD14+ monocytes. Cells were cultured in the presence of  granulocyte-monocyte colonies stimulating factor (GM-CSF) and IL-4 to obtain immature dendritic cells, wich were further cultured with PGE2 plus TNFa to induce their maturation. We obtained immature dendritic cells at  day 5 and mature dendritic cells at day 7, according to microscopic cellularphenotype and positivity of membrane markers for CD14, CD86, HLA-DR and CD83

GENERACIÓN in vitro DE CÉLULAS DENDRÍTICAS HUMANAS DE ORIGEN MIELOIDE

Dendritic cell activity is an important step to elicit cellular immunity. These cells are the most potent antigen presenting cells and the main activating factor for naive T cells. There are several phenotypes of dendritic cells originated from different cell precursors. The aim of this study was to assess a model to obtain human myeloid dendritic cells from peripheral blood CD14+ monocytes. Cells were cultured in the presence of  granulocyte-monocyte colonies stimulating factor (GM-CSF) and IL-4 to obtain immature dendritic cells, wich were further cultured with PGE2 plus TNFa to induce their maturation. We obtained immature dendritic cells at  day 5 and mature dendritic cells at day 7, according to microscopic cellularphenotype and positivity of membrane markers for CD14, CD86, HLA-DR and CD83

BIOLOGÍA DE LAS CÉLULAS DENDRÍTICAS HUMANAS

Antigen presenting cells are capable of capturing and processing antigenic peptides to be presented to T lymphocytes in the context of the Major Histocompatibility Complex. These peptides derived from self or no self proteins could be recognized  by the T Cell Receptor expressed on the surface of T lymphocytes. However, this presentation is not enough to activate peptide specific T lymphocytes. Dendritic cells are potent antigen presenting cells and  also have the capability to generate a second signal to stimulate T cell response. Currently, dendritic cells seem to play a central role in the  development of autoimmune diseases and also in the immune response  to microbial and tumoral antigens. The active study of their function and the possibility to purify dendritic cells form peripheral blood and even differentiate them in vitro from blood precursors will allow future and new  immune therapies for several diseases

BIOLOGÍA DE LAS CÉLULAS DENDRÍTICAS HUMANAS

Antigen presenting cells are capable of capturing and processing antigenic peptides to be presented to T lymphocytes in the context of the Major Histocompatibility Complex. These peptides derived from self or no self proteins could be recognized  by the T Cell Receptor expressed on the surface of T lymphocytes. However, this presentation is not enough to activate peptide specific T lymphocytes. Dendritic cells are potent antigen presenting cells and  also have the capability to generate a second signal to stimulate T cell response. Currently, dendritic cells seem to play a central role in the  development of autoimmune diseases and also in the immune response  to microbial and tumoral antigens. The active study of their function and the possibility to purify dendritic cells form peripheral blood and even differentiate them in vitro from blood precursors will allow future and new  immune therapies for several diseases

Comparación de la síntesis de interleucina1â por monocitos y linfocitos B estimulados con lipopolisacárido en pacientes con enfermedad periodontal

Background: Interleukin-1â (IL-1â) is considered an important mediator of bone loss in patients with periodontal disease. It is unclear whether there are differences in the production of IL-1â ex vivo by B cells and monocytes from peripheral blood between healthy patients and with periodontal disease. Objective: To quantify the production of IL-1â by monocytes and peripheral blood B cells of individuals with and without periodontal disease in the presence of lipopolysaccharide (LPS). Methods: Monocytes and peripheral blood B cells from 5 healthy/gingivitis, 5 chronic periodontitis (CP) and 5 aggressive periodontitis (AgP) patients were purified with anti-CD14 and anti-CD19 marked with magnetic beads. The purity of the cell populations was confirmed by flow citometry. Cells were stimulated with LPS and the production of IL-1â was quantified by ELISA. Results: Basal levels of IL-1â pr oduced by B lymphocytes wer e lower in subjects with chronic periodontitis than in healthy (p = 0.0079) and aggressive periodontitis patients (p = 0.03). In presence of LPS, the IL-1â levels are increased with no differences between groups. IL-1â produced by monocytes at baseline and with LPS, was similar between subjects, but higher compared to that released by B lymphocytes (p = 0.0079 in chronic periodontitis, p = 0.00571 in aggressive periodontitis) in the periodontitis groups. Conclusions: The production of IL-1â by LB at baseline is significantly reduced in CP patients compared to healthy and AgP, but once stimulated with LPS, the response is similar in the three groups. The ability of LB to produce IL-1â in response to stimulation with LPS is lower compared with monocytes.Antecedentes: la interleucina-1�À- (IL-1�À) se considera un mediador en la perdida osea en enfermedad periodontal. No es claro si existen diferencias en la produccion de IL-1�À ex vivo por linfocitos B (LB) y monocitos (Mo) de sangre periferica entre pacientes sanos y con enfermedad periodontal. Objetivo: cuantificar la produccion de IL-1�À por LB y Mo de sangre periferica de individuos con enfermedad periodontal y sin esta en presencia de lipopolisacarido (LPS). Metodos: se tomaron celulas de cinco individuos con diagnostico sano/gingivitis, cinco con periodontitis cronica (PC) y cinco con periodontitis agresiva (PAg). Los Mo y LB de sangre periferica se purificaron con anticuerpos anti-CD14 y anti-CD19 marcados con perlas magneticas. La pureza de las poblaciones celulares fue confirmada por citometria de flujo. Las celulas se estimularon con LPS y la produccion de IL-1�À se cuantifico por ELISA. Resultados: las concentraciones basales de IL-1�À producidas por LB fueron menores en sujetos con PC comparados con sujetos con PAg (p = 0,03) e individuos sanos (p = 0,0079). En presencia del estimulo, se incrementaron sin encontrarse diferencias entre los grupos. La IL-1�À producida por los Mo a nivel basal y frente al estimulo fue similar entre los sujetos, pero mayor comparada con la liberada por los LB (p = 0,0079 en PC y p = 0,00571 en PAg) en pacientes con periodontitis. Conclusiones: la produccion de IL-1�À por LB en el estado basal esta significativamente disminuida en pacientes con PC, comparado con sanos y PAg, pero una vez estimulados con LPS, la respuesta es similar en los tres grupos. La capacidad de los LB para producir IL-1�À en respuesta al estimulo con LPS es limitada comparada con la de los Mo

Comparación de la síntesis de interleucina-1β por monocitos y linfocitos B estimulados con lipopolisacárido en pacientes con enfermedad periodontal / Interleukin-1β Production by Peripheral Blood Monocytes and B Lymphocytes in Patients with [...]

Antecedentes: la interleucina-1β (IL-1β) se considera un mediador en la pérdida ósea en enfermedad periodontal. No es claro si existen diferencias en la producción de IL-1β ex vivo por linfocitos B (LB) y monocitos (Mo) de sangre periférica entre pacientes sanos y con enfermedad periodontal. Objetivo: cuantificar la producción de IL-1β por LB y Mo de sangre periférica de individuos con enfermedad periodontal y sin esta en presencia de lipopolisacárido (LPS). Métodos: se tomaron células de cinco individuos con diagnóstico sano/gingivitis, cinco con periodontitis crónica (PC) y cinco con periodontitis agresiva (PAg). Los Mo y LB de sangre periférica se purificaron con anticuerpos anti-CD14 y anti-CD19 marcados con perlas magnéticas. La pureza de las poblaciones celulares fue confirmada por citometría de flujo. Las células se estimularon con LPS y la producción de IL-1β se cuantificó por ELISA. Resultados: las concentraciones basales de IL-1β producidas por LB fueron menores en sujetos con PC comparados con sujetos con PAg (p = 0,03) e individuos sanos (p = 0,0079). En presencia del estímulo, se incrementaron sin encontrarse diferencias entre los grupos. La IL-1β producida por los Mo a nivel basal y frente al estímulo fue similar entre los sujetos, pero mayor comparada con la liberada por los LB (p = 0,0079 en PC y p = 0,00571 en PAg) en pacientes con periodontitis. Conclusiones: la producción de IL-1β por LB en el estado basal está significativamente disminuida en pacientes con PC, comparado con sanos y PAg, pero una vez estimulados con LPS, la respuesta es similar en los tres grupos. La capacidad de los LB para producir IL-1β en respuesta al estímulo con LPS es limitada comparada con la de los Mo. Background: Interleukin-1β (IL-1β) is considered an important mediator of bone loss in patients with periodontal disease. It is unclear whether there are differences in the production of IL-1β ex vivo by B cells and monocytes from peripheral blood between healthy patients and with periodontal disease. Objective: To quantify the production of IL-1β by monocytes and peripheral blood B cells of individuals with and without periodontal disease in the presence of lipopolysaccharide (LPS). Methods: Monocytes and peripheral blood B cells from 5 healthy/gingivitis, 5 chronic periodontitis (CP) and 5 aggressive periodontitis (AgP) patients were purified with anti-CD14 and anti-CD19 marked with magnetic beads. The purity of the cell populations was confirmed by flow citometry. Cells were stimulated with LPS and the production of IL-1β was quantified by ELISA. Results: Basal levels of IL-1β produced by B lymphocytes were lower in subjects with chronic periodontitis than in healthy (p = 0.0079) and aggressive periodontitis patients (p = 0.03). In presence of LPS, the IL-1β levels are increased with no differences between groups. IL-1β produced by monocytes at baseline and with LPS, was similar between subjects, but higher compared to that released by B lymphocytes (p = 0.0079 in chronic periodontitis, p = 0.00571 in aggressive periodontitis) in the periodontitis groups. Conclusions: The production of IL-1β by LB at baseline is significantly reduced in CP patients compared to healthy and AgP, but once stimulated with LPS, the response is similar in the three groups. The ability of LB to produce IL-1β in response to stimulation with LPS is lower compared with monocytes

Comparación de la síntesis de interleucina-1β por monocitos y linfocitos B estimulados con lipopolisacárido en pacientes con enfermedad periodontal / Interleukin-1β Production by Peripheral Blood Monocytes and B Lymphocytes in Patients with [...]

Antecedentes: la interleucina-1β (IL-1β) se considera un mediador en la pérdida ósea en enfermedad periodontal. No es claro si existen diferencias en la producción de IL-1β ex vivo por linfocitos B (LB) y monocitos (Mo) de sangre periférica entre pacientes sanos y con enfermedad periodontal. Objetivo: cuantificar la producción de IL-1β por LB y Mo de sangre periférica de individuos con enfermedad periodontal y sin esta en presencia de lipopolisacárido (LPS). Métodos: se tomaron células de cinco individuos con diagnóstico sano/gingivitis, cinco con periodontitis crónica (PC) y cinco con periodontitis agresiva (PAg). Los Mo y LB de sangre periférica se purificaron con anticuerpos anti-CD14 y anti-CD19 marcados con perlas magnéticas. La pureza de las poblaciones celulares fue confirmada por citometría de flujo. Las células se estimularon con LPS y la producción de IL-1β se cuantificó por ELISA. Resultados: las concentraciones basales de IL-1β producidas por LB fueron menores en sujetos con PC comparados con sujetos con PAg (p = 0,03) e individuos sanos (p = 0,0079). En presencia del estímulo, se incrementaron sin encontrarse diferencias entre los grupos. La IL-1β producida por los Mo a nivel basal y frente al estímulo fue similar entre los sujetos, pero mayor comparada con la liberada por los LB (p = 0,0079 en PC y p = 0,00571 en PAg) en pacientes con periodontitis. Conclusiones: la producción de IL-1β por LB en el estado basal está significativamente disminuida en pacientes con PC, comparado con sanos y PAg, pero una vez estimulados con LPS, la respuesta es similar en los tres grupos. La capacidad de los LB para producir IL-1β en respuesta al estímulo con LPS es limitada comparada con la de los Mo. Background: Interleukin-1β (IL-1β) is considered an important mediator of bone loss in patients with periodontal disease. It is unclear whether there are differences in the production of IL-1β ex vivo by B cells and monocytes from peripheral blood between healthy patients and with periodontal disease. Objective: To quantify the production of IL-1β by monocytes and peripheral blood B cells of individuals with and without periodontal disease in the presence of lipopolysaccharide (LPS). Methods: Monocytes and peripheral blood B cells from 5 healthy/gingivitis, 5 chronic periodontitis (CP) and 5 aggressive periodontitis (AgP) patients were purified with anti-CD14 and anti-CD19 marked with magnetic beads. The purity of the cell populations was confirmed by flow citometry. Cells were stimulated with LPS and the production of IL-1β was quantified by ELISA. Results: Basal levels of IL-1β produced by B lymphocytes were lower in subjects with chronic periodontitis than in healthy (p = 0.0079) and aggressive periodontitis patients (p = 0.03). In presence of LPS, the IL-1β levels are increased with no differences between groups. IL-1β produced by monocytes at baseline and with LPS, was similar between subjects, but higher compared to that released by B lymphocytes (p = 0.0079 in chronic periodontitis, p = 0.00571 in aggressive periodontitis) in the periodontitis groups. Conclusions: The production of IL-1β by LB at baseline is significantly reduced in CP patients compared to healthy and AgP, but once stimulated with LPS, the response is similar in the three groups. The ability of LB to produce IL-1β in response to stimulation with LPS is lower compared with monocytes