RESUMO
Disease caused by the new coronavirus (COVID-19) is characterized by fever, cough, and affection of the lower respiratory tract. It is associated with age, comorbidities and a weakened immune system. Typically, lymphopenias have been evidenced in severe cases and an excessive production of inflammatory cytokines (cytokine storm), which would explain the role of the hyperinflammatory response in the pathogenesis of COVID-19. Secondary inflammatory responses from virus reinfections may induce antibody-dependent enhancement (ADE), a viremic phenomenon that may be an alternative mechanism of cellular infection and should be considered when designing vaccines or immunotherapies involving the stimulation of neutralizing antibodies or the use of monoclonal antibodies. Currently, no vaccines or treatments demonstrate safety and efficacy in patients with COVID-19. However, the results from phase III clinical trials which involve the application of an mRNA (messenger ribonucleic acid) nucleic acid vaccine and an antiviral drug (remdisivir), are yet to be concluded. For the time being, the best measure to prevent the spread of COVID-19 is by implementing social isolation, this measure has been adopted by several countries as recommended by the World Health Organization (WHO).
La enfermedad causada por el nuevo coronavirus (COVID-19) se caracteriza por presentar fiebre y tos, afectar el tracto respiratorio inferior y estar asociada con la edad, comorbilidades y un sistema inmune debilitado. Típicamente se ha evidenciado linfopenias en los casos graves y una desmedida producción de citocinas inflamatorias (tormenta de citocinas), lo que explicaría el rol de la respuesta hiperinflamatoria en la patogénesis de la COVID-19. Las respuestas inflamatorias secundarias por reinfecciones del virus podrían inducir el aumento o la mejora dependiente de anticuerpos (ADE, por sus siglas en inglés), un fenómeno virémico que podría ser un mecanismo alternativo de infección celular y que se deberá tener en cuenta cuando se diseñen vacunas o inmunoterapias que involucren el estímulo de anticuerpos neutralizantes o el uso de anticuerpos monoclonales. Actualmente no existen vacunas ni tratamientos que demuestren seguridad y eficacia en pacientes con COVID-19; sin embargo, se espera la conclusión de los resultados de la aplicación de una vacuna de ácidos nucleicos ARNm (mensajero del ácido ribonucleico) y de un fármaco antiviral (remdisivir) que se encuentran en ensayos clínicos fase III. Por el momento la mejor medida para evitar la propagación de la infección es el aislamiento social exhaustivo y viene siendo adoptado por varios países según recomendación de la Organización Mundial de la Salud (OMS).
Assuntos
Infecções por Coronavirus/epidemiologia , Pneumonia Viral/epidemiologia , Isolamento Social , Antivirais/administração & dosagem , COVID-19 , Infecções por Coronavirus/tratamento farmacológico , Infecções por Coronavirus/imunologia , Infecções por Coronavirus/prevenção & controle , Citocinas/imunologia , Humanos , Pandemias/prevenção & controle , Pneumonia Viral/imunologia , Pneumonia Viral/prevenção & controle , Fatores de Risco , Vacinas Virais/administração & dosagem , Tratamento Farmacológico da COVID-19RESUMO
RESUMEN La enfermedad causada por el nuevo coronavirus (COVID-19) se caracteriza por presentar fiebre y tos, afectar el tracto respiratorio inferior y estar asociada con la edad, comorbilidades y un sistema inmune debilitado. Típicamente se ha evidenciado linfopenias en los casos graves y una desmedida producción de citocinas inflamatorias (tormenta de citocinas), lo que explicaría el rol de la respuesta hiperinflamatoria en la patogénesis de la COVID-19. Las respuestas inflamatorias secundarias por reinfecciones del virus podrían inducir el aumento o la mejora dependiente de anticuerpos (ADE, por sus siglas en inglés), un fenómeno virémico que podría ser un mecanismo alternativo de infección celular y que se deberá tener en cuenta cuando se diseñen vacunas o inmunoterapias que involucren el estímulo de anticuerpos neutralizantes o el uso de anticuerpos monoclonales. Actualmente no existen vacunas ni tratamientos que demuestren seguridad y eficacia en pacientes con COVID-19; sin embargo, se espera la conclusión de los resultados de la aplicación de una vacuna de ácidos nucleicos ARNm (mensajero del ácido ribonucleico) y de un fármaco antiviral (remdisivir) que se encuentran en ensayos clínicos fase III. Por el momento la mejor medida para evitar la propagación de la infección es el aislamiento social exhaustivo y viene siendo adoptado por varios países según recomendación de la Organización Mundial de la Salud (OMS).
ABSTRACT Disease caused by the new coronavirus (COVID-19) is characterized by fever, cough, and affection of the lower respiratory tract. It is associated with age, comorbidities and a weakened immune system. Typically, lymphopenias have been evidenced in severe cases and an excessive production of inflammatory cytokines (cytokine storm), which would explain the role of the hyperinflammatory response in the pathogenesis of COVID-19. Secondary inflammatory responses from virus reinfections may induce antibody-dependent enhancement (ADE), a viremic phenomenon that may be an alternative mechanism of cellular infection and should be considered when designing vaccines or immunotherapies involving the stimulation of neutralizing antibodies or the use of monoclonal antibodies. Currently, no vaccines or treatments demonstrate safety and efficacy in patients with COVID-19. However, the results from phase III clinical trials which involve the application of an mRNA (messenger ribonucleic acid) nucleic acid vaccine and an antiviral drug (remdisivir), are yet to be concluded. For the time being, the best measure to prevent the spread of COVID-19 is by implementing social isolation, this measure has been adopted by several countries as recommended by the World Health Organization (WHO).
Assuntos
Humanos , Pneumonia Viral/epidemiologia , Isolamento Social , Infecções por Coronavirus/epidemiologia , Antivirais/administração & dosagem , Pneumonia Viral/imunologia , Pneumonia Viral/prevenção & controle , Vacinas Virais/administração & dosagem , Fatores de Risco , Citocinas/imunologia , Infecções por Coronavirus/imunologia , Infecções por Coronavirus/prevenção & controle , Infecções por Coronavirus/tratamento farmacológico , Pandemias/prevenção & controle , COVID-19RESUMO
RESUMEN En los últimos años, las células troncales mesenquimales (Mesenchymal Stem Cells, MSC) han adquirido mucha importancia debido a su gran plasticidad y su capacidad de liberar factores paracrinos con capacidad de interactuar con diversos tipos celulares, tejidos y órganos. El uso de MSC en medicina regenerativa es importante debido a que, al no expresar las moléculas del complejo mayor de histocompatibilidad (MHC) clase II ni moléculas co-estimuladoras y tener baja expresión del MHC clase I, haría que no sean rechazadas por individuos de la misma especie, es posible utilizarlas no sólo de manera autóloga, sino también, eventualmente, alogénica. Sin embargo, es importante demostrar científicamente muchas de sus propiedades, entre ellas las inmunomoduladoras. Al tener varias fuentes de obtención, se debe estandarizar la que sea la mejor para garantizar la pureza y calidad de las MSC. Finalmente, es importante que cuando se trabaje con estas células se demuestre completamente las características del cultivo celular, la inmunotipificación y su capacidad de diferenciación. Se están ensayando muchas aplicaciones clínicas de las MSC. Dentro de ellas, su capacidad para mejorar la recuperación y potencial curación de úlceras crónicas como las diabéticas, ha atraído la atención por su potencial impacto terapéutico.
ABSTRACT In recent years, mesenchymal stem cells (MSC) have become very important due to their high plasticity and their ability to release paracrine factors able to interact with various cell types, tissues and organs. The use of MSC in regenerative medicine became of vital importance, since they do not express histocompatibility MHC molecules class II nor costimulant molecules, and low expression of MHC class I, will not be rejected by individuals of same species, they could be used in an autologous, and eventually, allogeneic manner. However, it is important to scientifically demonstrate many properties, including immunomodulatory ones. Having several sources of obtaining, it should be standardized the best one to ensure the purity and quality of these cells. Finally, it is important when working with these cells, that characteristics of cell culture, immunophenotyping and differentiation capacity are fully demonstrated. MSC have been applied in several clinical uses. Among them, their ability to improve, and even heal chronic ulcers, as diabetic, has attracted attention for its potential therapeutic impact.
Assuntos
Humanos , Células-Tronco Mesenquimais , Transplante de Células-Tronco MesenquimaisRESUMO
In recent years, mesenchymal stem cells (MSC) have become very important due to their high plasticity and their ability to release paracrine factors able to interact with various cell types, tissues and organs. The use of MSC in regenerative medicine became of vital importance, since they do not express histocompatibility MHC molecules class II nor costimulant molecules, and low expression of MHC class I, will not be rejected by individuals of same species, they could be used in an autologous, and eventually, allogeneic manner. However, it is important to scientifically demonstrate many properties, including immunomodulatory ones. Having several sources of obtaining, it should be standardized the best one to ensure the purity and quality of these cells. Finally, it is important when working with these cells, that characteristics of cell culture, immunophenotyping and differentiation capacity are fully demonstrated. MSC have been applied in several clinical uses. Among them, their ability to improve, and even heal chronic ulcers, as diabetic, has attracted attention for its potential therapeutic impact.
Assuntos
Células-Tronco Mesenquimais , Humanos , Transplante de Células-Tronco MesenquimaisRESUMO
Evaluar el efecto inmunomodulador sobre poblaciones linfocitarias, células dendríticas (DC), citoquinas Th1/Th2/Th17 (T-helper) e inflamatorias en el ámbito sistémico y/o en el microambiente tumoral de ratones con o sin melanoma. Materiales y métodos. Se obtuvieron muestras de sangre periférica y/o de tumores primarios de ratones con melanoma B16 tratados o no con un extracto hidroalcohólico de Uncaria tomentosa (UT) con 5,03% de alcaloides oxindólicos pentacíclicos (UT-POA) obtenido de la corteza de la planta. Todos los ensayos de medición de células y citoquinas fueron realizados por citometría de flujo. Resultados. UT-POA a nivel sistémico incrementa la relación CD4/CD8a (Cluster of Differenciation), mientras que la activación celular es inversamente proporcional; incrementa la proporción de DCm (DC mieloides); induce un perfil Th1 proinflamatorio y reduce la respuesta Th17. TNF-a (tumor necrosis factor alpha) y IL-17A (interleuquina) correlacionan positiva y negativamente con la relación CD4/CD8a. Conclusiones. El incremento de Th1 (TNF-a) puede tener como consecuencia el incremento de linfocitos CD4 o la activación de macrófagos M1. Aunque UT-POA muestra un incremento de DCm, este no es dosis-dependiente. La disminución de Th17 (IL-17A) puede favorecer el funcionamiento de los linfocitos CD8a. UT-POA muestra mejores efectos inmunomoduladores en el ámbito sistémico que intratumoral...
To evaluate the immunomodulatory effect on lymphocyte subsets, dendritic cells (DC), Th1 / Th2 / Th17 and inflammatory cytokines on systemic level and/or in the tumor microenvironment of mice with or without melanoma. Materials and methods: Peripheral blood and/or primary tumors samples were obtained of mice with B16 melanoma treated or not with a hydroalcoholic extract of Uncaria tomentosa (UT) with 5.03% of pentacyclic oxindole alkaloids (UT-POA) obtained from the bark of the plant. All cell assays and cytokine measurements were performed by flow cytometry. Results. UT-POA systemically increased CD4/CD8a relation while cell activation was inversely proportional; increased the proportion of DCm; induced a pro-inflammatory Th1 profile and reduced Th17 response. TNF-a and IL-17A positively and negatively correlated with CD4/CD8a relation. Conclusions. The increase of Th1 (TNF-a) may result in the increase of CD4 or M1 macrophage activation. Although UT-POA shows increased DCm, is not dose-dependent. Th17(IL-17A) decreased can support the function of CD8a lymphocytes. UT-POA shows better systemic immunomodulatory effects than intratumoral...
Assuntos
Animais , Camundongos , Ativação Linfocitária , Células Dendríticas , Unha-de-GatoRESUMO
Evaluar el efecto inmunomodulador del extracto estandarizado (5,03%, alcaloides oxindólicos pentacíclicos) de Uncaria tomentosa (UT-POA) sobre el inmunofenotipo de células dendríticas (DC) y IL-12/Th1/Th2/Th17 en pacientes con cáncer de mama estadio II (BCII) y mujeres sanas (H). Materiales y métodos. Se obtuvó sangre de 11 H y 7 BCII, se aislaron y cultivaron PBMC por 2 h con/sin distintas concentraciones de UT-POA y se estimularon o no con LPS por 24 h. Las PBMC fueron marcadas con anticuerpos específicos para DC y en el sobrenadante se midió las citoquinas Th1/Th2/Th17, ambos por citometría de flujo. Además, se midió IL-12 por ELISA. Resultados. UT-POA no alteró a las DC o la expresión de moléculas accesorias en BCII. Sin embargo, en H mostró una disminución en el porcentaje de DC mieloides (mDC) con incremento de HLA-DR y CD86 a 1000 ug/mL. La secreción de IL-12 fue modificada solo en H, incrementando la subunidad p70 y disminuyendo la p40. UT-POA incremento Th1 (IFN-y y IL-2), Th2 (IL-4) y Th17 (IL-17) en BCII y H. Conclusiones. UT-POA incrementa la producción de citoquinas relacionadas con la respuesta antitumoral a concentraciones entre el rango de 500-1000 ug/mL. Este efecto positivo debería ser evaluado no solo sistemicamente sino también en el microambiente tumoral. La UT-POA puede ser un fitoquímico útil en la quimioprevención y en el uso alternativo con terapias contra el cáncer...
This study aimed to research the in vitro immunomodulatory effects of an Uncaria tomentosa hydroalcoholic extract standardized (5.03%, pentacyclic oxindole alkaloids) (UT-POA) on the immunophenotype of dendritic cells (DC) subsets, Th1, Th2, Th17 and IL-12 cytokines from patients with stage II breast cancer (BCII) and healthy women (H). Materials and methods. Blood of 11 H and 7 BCII was obtained, PBMC were isolated and cultured for 2h with/without various concentrations of UT-POA and stimulated or not with LPS for 24h. PBMC were labeled with specific antibodies for DC and in the supernatant we measured Th1/Th2/Th17 cytokines, both by flow cytometry. Furthermore IL-12 was measured by ELISA. Results. UT-POA did not alter DC or accessory molecules expression in BCII. However, H exhibited a decrease in the percentage of mDC (myeloid DC) and an increase in HLA-DR and CD86 expression at 1000 ug/mL. IL-12 secretion was modified only in the H group, increasing p70 subunit and decreasing p40 subunit. UT-POA increased Th1 (IFN-y and IL-2), Th2 (IL-4) and Th17 (IL-17) secretion in both groups. Conclusions. UT-POA increased the production of cytokines related with anti-tumoral response at concentrations of 500-1000 ug/mL. This positive effect should be evaluated not only systemically but also in the tumor microenvironment in further studies. UT-POA may be a useful phytochemical in chemoprevention and in the alternative use in cancer therapies...
Assuntos
Humanos , Adolescente , Adulto , Feminino , Criança , Adulto Jovem , Pessoa de Meia-Idade , Anti-Inflamatórios , Células Dendríticas , Neoplasias da Mama , Unha-de-GatoRESUMO
This is a narrative review that shows accessible information to the scientific community about melanoma and immunotherapy. Dendritic cells have the ability to participate in innate and adaptive immunity, but are not unfamiliar to the immune evasion of tumors. Knowing the biology and role has led to generate in vitro several prospects of autologous cell vaccines against diverse types of cancer in humans and animal models. However, given the low efficiency they have shown, we must implement strategies to enhance their natural capacity either through the coexpression of key molecules to activate or reactivate the immune system, in combination with biosimilars or chemotherapeutic drugs. The action of natural products as alternative or adjuvant immunostimulant should not be ruled out. All types of immunotherapy should measure the impact of myeloid suppressor cells, which can attack the immune system and help tumor progression, respectively. This can reduce the activity of cellular vaccines and/or their combinations, that could be the difference between success or not of the immunotherapy. Although for melanoma there exist biosimilars approved by the Food and Drug Administration (FDA), not all have the expected success. Therefore it is necessary to evaluate other strategies including cellular vaccines loaded with tumor antigenic peptides expressed exclusively or antigens from tumor extracts and their respective adjuvants.
Assuntos
Vacinas Anticâncer/uso terapêutico , Células Dendríticas , Imunoterapia , Melanoma/tratamento farmacológico , Neoplasias Cutâneas/tratamento farmacológico , Animais , Medicamentos Biossimilares , Modelos Animais de Doenças , HumanosRESUMO
La presente es una revisión narrativa que muestra la información accesible a la comunidad científica sobre melanoma e inmunoterapia. Las células dendríticas tienen la capacidad de participar en la inmunidad innata y adaptativa, pero no son ajenas a la evasión inmune de los tumores. Conocer su biología y rol ha llevado a generar in vitro varios prospectos de vacunas celulares autólogas contra diversos tipos de cáncer en humanos y modelos animales; sin embargo, en vista de la poca eficiencia que han mostrado, se deben implementar estrategias para potenciar su capacidad natural ya sea a través de la coexpresión de moléculas clave para activar o reactivar al sistema inmune, en combinación con biosimilares o drogas quimioterapeúticas y no se debe descartar la acción de productos naturales como alternativa inmunoestimulante o adyuvante. Todos los tipos de inmunoterapía deberían medir el impacto de las células supresoras de origen mieloide, las que pueden atacar al sistema inmune y ayudar a la progresión tumoral, respectivamente. Esto puede reducir la actividad de las vacunas celulares y/o sus combinaciones pudiendo ser la diferencia entre el éxito o no de la inmunoterapia. Aunque en melanoma existen biosimilares aprobados por la Food and Drug Administration (FDA) no todos tienen el éxito esperado por lo que es necesario evaluar otras estrategias que incluyan vacunas celulares cargadas con péptidos antigénicos tumorales expresados exclusivamente o antígenos provenientes de extractos tumorales y sus respectivos adyuvantes...
This is a narrative review that shows accessible information to the scientific community about melanoma and immunotherapy. Dendritic cells have the ability to participate in innate and adaptive immunity, but are not unfamiliar to the immune evasion of tumors. Knowing the biology and role has led to generate in vitro several prospects of autologous cell vaccines against diverse types of cancer in humans and animal models. However, given the low efficiency they have shown, we must implement strategies to enhance their natural capacity either through the coexpression of key molecules to activate or reactivate the immune system, in combination with biosimilars or chemotherapeutic drugs. The action of natural products as alternative or adjuvant immunostimulant should not be ruled out. All types of immunotherapy should measure the impact of myeloid suppressor cells, which can attack the immune system and help tumor progression, respectively. This can reduce the activity of cellular vaccines and/or their combinations, that could be the difference between success or not of the immunotherapy. Although for melanoma there exist biosimilars approved by the Food and Drug Administration (FDA), not all have the expected success. Therefore it is necessary to evaluate other strategies including cellular vaccines loaded with tumor antigenic peptides expressed exclusively or antigens from tumor extracts and their respective adjuvants...
Assuntos
Humanos , Células Dendríticas/imunologia , Imunoterapia , Melanoma , Unha-de-Gato/imunologia , VacinasRESUMO
OBJECTIVES: To evaluate the immunomodulatory effect on lymphocyte subsets, dendritic cells (DC), Th1 / Th2 / Th17 and inflammatory cytokines on systemic level and/or in the tumor microenvironment of mice with or without melanoma. MATERIALS AND METHODS: Peripheral blood and/or primary tumors samples were obtained of mice with B16 melanoma treated or not with a hydroalcoholic extract of Uncaria tomentosa (UT) with 5.03% of pentacyclic oxindole alkaloids (UT-POA) obtained from the bark of the plant. All cell assays and cytokine measurements were performed by flow cytometry. RESULTS: UT-POA systemically increased CD4/CD8a relation while cell activation was inversely proportional; increased the proportion of DCm; induced a pro-inflammatory Th1 profile and reduced Th17 response. TNF-α and IL-17A positively and negatively correlated with CD4/CD8a relation. CONCLUSIONS: The increase of Th1 (TNF-α) may result in the increase of CD4 or M1 macrophage activation. Although UT-POA shows increased DCm, is not dose-dependent. Th17(IL-17A) decreased can support the function of CD8a lymphocytes. UT-POA shows better systemic immunomodulatory effects than intratumoral.
Assuntos
Unha-de-Gato , Células Dendríticas , Subpopulações de Linfócitos , Melanoma/tratamento farmacológico , Extratos Vegetais/farmacologia , Neoplasias Cutâneas/tratamento farmacológico , Animais , Citocinas , CamundongosRESUMO
UNLABELLED: Objetives. This study aimed to research the in vitro immunomodulatory effects of an Uncaria tomentosa hydroalcoholic extract standardized (5.03%, pentacyclic oxindole alkaloids) (UT-POA) on the immunophenotype of dendritic cells (DC) subsets, Th1, Th2, Th17 and IL-12 cytokines from patients with stage II breast cancer (BCII) and healthy women (H). MATERIALS AND METHODS: Blood of 11 H and 7 BCII was obtained, PBMC were isolated and cultured for 2h with/without various concentrations of UT-POA and stimulated or not with LPS for 24h. PBMC were labeled with specific antibodies for DC and in the supernatant we measured Th1/Th2/Th17 cytokines, both by flow cytometry. Furthermore IL-12 was measured by ELISA. RESULTS: UT-POA did not alter DC or accessory molecules expression in BCII. However, H exhibited a decrease in the percentage of mDC (myeloid DC) and an increase in HLA-DR and CD86 expression at 1000 µg/mL. IL-12 secretion was modified only in the H group, increasing p70 subunit and decreasing p40 subunit. UT-POA increased Th1 (IFN-γ and IL-2), Th2 (IL-4) and Th17 (IL-17) secretion in both groups. CONCLUSIONS: UT-POA increased the production of cytokines related with anti-tumoral response at concentrations of 500-1000 µg/mL. This positive effect should be evaluated not only systemically but also in the tumor microenvironment in further studies. UT-POA may be a useful phytochemical in chemoprevention and in the alternative use in cancer therapies.
Assuntos
Neoplasias da Mama/tratamento farmacológico , Unha-de-Gato , Células Dendríticas , Interleucina-12 , Extratos Vegetais/uso terapêutico , Neoplasias da Mama/imunologia , Feminino , Humanos , Imunomodulação , Leucócitos MononuclearesRESUMO
Lepidium meyenii (Maca) is a plant that grows at over 4,000 m above sea level in the central Peruvian Andes. The hypocotyls of this plant are traditionally consumed for their nutritional and medicinal properties. The aim of this study was to determine the health status based on a health related quality of life (HRQL) questionnaire (SF-20) and serum levels of interleukin 6 (IL-6) in subjects that are maca consumers. For this, a cross-sectional study was designed to be performed in 50 subjects from Junin (4,100 m): 27 subjects were maca consumers and 23 were non-consumers. The SF-20 survey is used to obtain a summary measure of health status. The stand up from a chair and sit down (SUCSD) test (to assess lower-extremity function), hemoglobin measurement, blood pressure, sexual hormone levels, serum IL-6 levels and the score of chronic mountain sickness (CMS) were evaluated. Testosterone/estradiol ratio (P <0.05), IL-6 (P < 0.05) and CMS score were lower, whereas the health status score was higher, in maca consumers when compared to non-consumers (P < 0.01). A greater proportion of maca consumers successfully completed the SUCSD test compared to non-consumers (P < 0.01), showing a significant association with lower values of serum IL-6 (P < 0.05). In conclusion, consumption of maca was associated with low serum IL-6 levels and in turn with better health status scores in the SF-20 survey and low chronic mountain sickness scores.
Assuntos
Doença da Altitude/prevenção & controle , Dieta , Interleucina-6/sangue , Lepidium , Aptidão Física , Fitoterapia , Preparações de Plantas/uso terapêutico , Adulto , Idoso , Altitude , Estudos Transversais , Estradiol/sangue , Feminino , Nível de Saúde , Inquéritos Epidemiológicos , Humanos , Hipocótilo , Extremidade Inferior , Masculino , Pessoa de Meia-Idade , Peru , Preparações de Plantas/farmacologia , Qualidade de Vida , Inquéritos e Questionários , Testosterona/sangueRESUMO
Objetivo. Determinar el efecto de un extracto hidroalcohólico de uña de gato, Uncaria tomentosa (UG) sobre célulasdendríticas (DC) de sangre periférica y la expresión de HLA-DR y CD86 en muestras de sangre periférica de individuos sanos tratadas con lipopolisacáridos (LPS). Materiales y métodos. El polvo de la corteza de UG se preparó como una decocción estéril a 30g/L por 30 minutos. Se obtuvo muestras de sangre periférica de individuos sanos. Las célulasmononucleares de sangre periférica (CMSP) fueron separadas por gradiente de centrifugación, pretratadas o no durante dos horas con distintas concentraciones de UG y estimuladas 24 h con LPS, luego fueron marcadas con anticuerpos monoclonales fluorescentes específicos para HLA-DR, Linaje tipo 1 (Lin1), CD11c y CD86 y preparadas para la citometría de flujo. Resultados. Comparando con el grupo de CMSP con LPS pero sin UG se observó, de maneradosis dependiente, una disminución en el porcentaje de DC mieloides (DCm) (p menor que 0,05) y una tendencia a incrementar el porcentaje de DC plasmacitoides (DCp). En las DCp se observó una disminución de la intensidad de fluorescencia media (IFM) de HLA-DR sólo a 500 y 1000 μg/mL de UG (p menor que 0,001); mientras que hubo un incremento de la IFM de CD86en todo el rango (p menor que 0,05). En DCm a las mismas concentraciones no se observó efecto de UG sobre la IFM de HLA-DR y CD86. Conclusiones. Este extracto estandarizado de UG tiende a incrementar el porcentaje de DCp y disminuye el porcentaje de DCm. UG no afecta la expresión de HLA-DR y CD86 en DCm. Este estudio demuestra que este extracto de UG favorece la activación/diferenciación de DCp, la cual participa en mecanismos de respuesta inmune adaptativa.
Objective. To determine the effect of an hydro-alcoholic extract of cat´s claw Uncaria tomentosa (UG) overlipopolysaccharides û treated dendritic cells (DC) and HLA-DR and CD86 molecules expression of peripheral bloodsamples obtained from healthy individuals. Materials and methods. Peripheral blood samples were collected from healthy individuals. Peripheral Blood Mononuclear Cells (PBMC) were isolated by centrifugation over density gradient, pre-treated with and without the addition of UG, and then stimulated for 24 h with lipopolysaccharides (LPS), then the cells were labelled with specific fluorescent monoclonal antibodies to HLA-DR, Linage type 1 (Lin1), CD11c and CD86 and prepared for flow. Results. Compared to PBMC with LPS but without UG group, we found a decrease of myeloid DC (mDC) percentage (p minor that 0.05) and a tendency to increase plasmacytoid DC (pDC) percentage, both in a dose-dependent manner. There was a decrease in media fluorescence intensity (MFI) of HLA-DR in DCp between 500 and 1000 μg/mL of CC (p minor that 0,001), but we found a increase in MFI of CD86 at all concentrations (p<0,05). There was no effect of UG overMFI of the HLA-DR or CD86 expression in mDC. Conclusions. UG showed a tendency to increase pDC percentage and decreased mDC percentage. UG did not affect the expression of CD86 and HLA-DR from mDC. This study demonstrates that UG promotes activation/differentiation of pDC, which has a demonstrated involvement in the adaptive immune response mechanisms.
Assuntos
Humanos , Células Dendríticas , Plantas Medicinais , Uncaria , Unha-de-GatoRESUMO
Introducción: la artritis reumatoide (AR) es una enfermedad autoinmune caracterizada por un proceso inflamatorio crónico articular. En los últimos años se está resaltando la importancia de las células dendríticas en AR, debido a su capacidad de presentar autoantígenos y estimular a linfocitos T autoreactivos. En este estudio se ensayó la capacidad de Uncaria tomentosa (Uña de Gato, UG), una planta peruana con propiedades inmunomoduladoras, sobre la población de células dendríticas circulantes y sobre la expresión de sus moléculas de maduración y coestimulación. Objetivos: determinar la población de células dendríticas (DC) de origen mieloide (DCm) y plasmocitoide (DCp) por citometría de flujo en pacientes con AR. Evaluar la variación en la expresión de moléculas HLA-DR y CD86 en ambas subpoblaciones celulares expuestas a diferentes concentraciones de un extracto hidroalcohólico de UG con 5 por ciento de alcaloides oxindólicos pentacíclicos (UG-POA). Material y método: se evaluaron DC a partir de muestras de sangre periférica de pacientes con AR y adultos controles sanos, enfrentadas a diferentes concentraciones de UG. Las muestras fueron marcadas con anticuerpos monoclonales específicos, evaluadas por citometría de flujo y analizadas con el software Summit 4.3. El análisis estadístico fue realizado mediante Test de rangos de Friedman para muestras relacionadas, Test T de student y Prueba de Tendencias y Análisis de Medidas Repetidas. Resultados: encontramos que UG-POA disminuyó de manera dosis dependiente la subpoblación de DCm de sangre periférica de pacientes con AR, sin afectar la subpoblación de DCp y aumentó la expresión de las moléculas HLA-DR y CD86 en DCm. Conclusiones: la UG-POA disminuye la subpoblación de DCm, mientras incrementa la expresión de moléculas HLA-DR y CD86 en pacientes con AR definida. Estos hallazgos añadirían un mecanismo adicional al efecto inmuno-modulador de la UG en procesos inflamatorios crónicos.
Introduction: Rheumatoid arthritis (RA) is an autoimmune disease characterized by a chronic inflammatory process with in the joints. Recently, the importance of dendritic cells (DC) has been emphasized in this condition due to their remarkable ability for presenting autoantigens and for stimulating auto-reactive T-cells. The ability of Uncariatomentosa (CatÆs claw), a native Peruvian plant with immune-modulating properties on the circulating dendritic cell population and on their maturation and co-stimulation capacities was assessed. Objectives: To determine subsets of myeloid and plasmacytoid dendritic cells (mDC and pDC, respectively) in patients with RA with the use of flow cytometry. To evaluate variations in the expression of HLA-DRand CD86 molecules in both subsets of DC in response to different concentrations of a hydro-alcoholic extract of Catïs claw with 5 per cent of pentacyclic oxindole alkaloids. Materials and methods: Peripheral blood dendritic cells were taken from RA patients and healthy controls, and they were incubated with different concentrations of Catïs claw. Then, cells were labeled with specific monoclonal antibodies and they were separated using flow cytometry and they were analyzed with a Summit 4.3. software. Statistical analysis was performed using Friedmanïs, T-student, trends and repeat measure analysis tests. Results: We found that Catïs claw extract decreased in a dose-dependent fashion mDC population in the peripheral blood of RA patients without affecting pDC, and it also increased HLA-DR and CD86 expression on mDC. Conclusions: Catïs claw reduced the mDC subset, while it increased HLA-DR and CD86 expression in RA patients. This is an additional mechanism for the known immune-modulating effect of Catïs claw in chronic inflammatory conditions.
Assuntos
Humanos , Artrite Reumatoide , Células Dendríticas , Unha-de-Gato , Unha-de-GatoRESUMO
Objetivo: Evaluar la acción antiinflamatoria de la capsaicina en la producción del factor de necrosis tumoral de tipo alfa (TNF menos alfa) en células mononucleares de sangre periférica de rata (CMSP), previamente estimuladas con lipopolisacáridos (LPS). Materiales y métodos: Las CMSP provinieron de ratas hembras de la cepa Sprague-Dawley de 8-14 semanas, se obtuvieron por gradiente de centrifugación y se estimularon o no con LPS (100 ng/mL) por 24 h considerándolas como grupos control positivo y negativo de la producción de TNF menos alfa;, respectivamente. Dos horas antes de la estimulación con LPS otros grupos de CMSP fueron tratados con capsaicina (pureza mínima de 97 por ciento, SIGMA) a concentraciones de 0,01 µM; 0,1 µM y 1µM (grupos de evaluación). Los niveles de TN menos alfa; se determinaron por ELISA en el sobrenadante del cultivo celular. El análisis estadístico se realizó por ANOVA y prueba de Tukey para comparaciones múltiples. Resultados: Los niveles de TNF menos alfa; a las concentraciones de capsaicina de 0,01 µM y 0,1 µM no mostraron cambios significativos respecto al control positivo; sin embargo, a 1µM se produjo una disminución de 21,9 por ciento (p<0,05) de esta citokina. Conclusión: La capsaicina evidencia un efecto antiinflamatorio al disminuir los niveles de TNF menos alfa; una citokina pro inflamatoria en un modelo in vitro en cultivos celulares
Objective: Anti-inflammatory actions of capsaicine for the production of tumor necrosis factor-alpha (TNF-á) in lipopolysaccharide (LPPS)-stimulated mononuclear cells from peripheral blood of rats. Material and methods: The aforementioned cells were obtained from Sprague-Dawley female rats, 8 to 14 weeks old, they were obtained using a centrifugation gradient and some of them were stimulated with LPPS (100 ng/mL), and some were not, so they were classified as positive and negative control groups according to TNF-á production. Two hours before stimulation with LPPS, other groups of mononuclear cells from peripheral blood of rats were treated with capsaicine (minimal purity, 97, SIGMA) in 0,01 ìM, 0,1 ìM, and 1 ìM concentrations (groups for assessment). TNF-á levels were determined using an ELISA test in the supernatant of the cell culture. Statistical tests were performed using ANOVA and Tukeys test for multiple comparisons. Results: TNF-á levels at 0,01 and 0,1 ìM capsaicine concentrations did not show significant changes with respect to positive controls; however, at 1 ìM there was a 21,9% reduction (p<0.05) in TNF-á levels. Conclusion: There is evidence showing that capsaicine exerts an anti-inflammatory effect when TNF-á levels are reduced, being the latter a pro-inflammatory cytokine in an in vitro model using cell cultures.