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1.
An. bras. dermatol ; 94(1): 52-55, Jan.-Feb. 2019. tab, graf
Article in English | LILACS | ID: biblio-983741

ABSTRACT

Abstract: Background: Pityriasis rosea is a common papulosquamous disorder. However, its etiology and pathogenesis remain unclear. Objective: We investigate the types of inflammatory cells infiltrating the lesional skin of pityriasis rosea and demonstrate whether T-cell-mediated immunity is involved in the pathogenesis of this condition or not. Methods: The biopsies were taken from the lesional skin of 35 cases of patients diagnosed with pityriasis rosea. The specimens were prepared in paraffin sections, then submitted to routine immunohistochemistry procedures using monoclonal antibodies directed against CD3, CD4, CD8, CD20 and CD45RO and horseradish peroxidase-labeled goat anti-human antibodies. The positive sections were determined by the ratio and staining intensity of positive inflammatory cells. Results: The mean score of positive CD3, CD4, CD8, and CD45RO staining was respectively 3.74±3.88, 5.67±4.40, 2.94±3.42 and 7.68±4.33 in these pityriasis rosea patients (P<0.001). The percentage of positive staining was 54.29% (19/35), 69.7% (23/33), 40% (14/35) and 79.41% (27/34) (P<0.05). However, the staining of CD20 was negative in all samples. The mean score of CD3 staining in patients with time for remission ≤60 days (4.90±4.21) was higher than that in patients with time for remission >60 days (2.00±2.5) (P<0.05), whereas no statistical difference in the mean score of CD4, CD8 and CD45RO staining was observed. study liMitations: The sample size and the selected monoclonal antibody are limited, so the results reflect only part of the cellular immunity in the pathogenesis of pityriasis rosea. Conclusion: Our findings support a predominantly T-cell mediated immunity in the development of pityriasis rosea.


Subject(s)
Humans , Male , Female , Adolescent , Adult , Middle Aged , Young Adult , T-Lymphocyte Subsets/pathology , Pityriasis Rosea/pathology , Reference Values , Staining and Labeling , Time Factors , Biopsy , Immunohistochemistry , CD4-Positive T-Lymphocytes/pathology , T-Lymphocyte Subsets/immunology , Pityriasis Rosea/immunology , Leukocyte Common Antigens/analysis , CD3 Complex/analysis , CD8-Positive T-Lymphocytes/pathology , Immunity, Cellular
2.
Rev. méd. Chile ; 146(2): 150-159, feb. 2018. tab, graf
Article in English | LILACS | ID: biblio-961372

ABSTRACT

ABSTRACT Background: The dual potential to promote tolerance or inflammation when facing self-antigens makes dendritic cells (DCs) fundamental players in autoimmunity. There is an association between smoking and DCs maturation in patients with rheumatoid arthritis (RA). However, ethnicity is a key factor in autoimmune disorders. Aim: To evaluate phenotypic and functional alterations of DCs obtained from Chilean patients with RA as compared to healthy controls (HC). In second term, to compare the inflammatory behaviour of DCs between smoker and non-smoker patients. Material and Methods: Monocyte-derived DCs and T-cells were obtained from blood samples isolated from 30 HC and 32 RA-patients, 14 of which were currently smokers and 18 non-smokers. Several maturation surface markers were evaluated in DCs, including HLA-DR, CD40, CD80, CD83 and CD86. Furthermore, autologous co-cultures of DCs and T-cells were carried out and then T-cell proliferation, and expansion of Th1, Th17 and Tregs were analysed. Results: Compared with HC, RA-patients displayed increased HLA-DR expression in DCs, which was manifested mainly in patients with moderate-to- high disease activity scores (DAS28). Furthermore, RA-patients presented a stronger Th17-expansion and a correlation between DAS28 and Th1-expansion. Both effects were mainly observed in patients in remission or with a low DAS28. Moreover, smoker RA-patients displayed enhanced HLA-DR and CD83 expression in DCs as well as an exacerbated Th17-expansion and a correlation between DAS28 and Th1-expansion. Conclusions: These findings suggest that smoking enhances the inflammatory behaviour of DCs and the consequent Th1 and Th17-mediated response in patients with RA


Introducción: El potencial dual que poseen para promover tolerancia o inflamación ante antígenos propios, hace de las células dendríticas (CDs) actores fundamentales en el desarrollo de autoinmunidad. Existe una asociación entre fumar y la maduración de las CDs en pacientes con artritis reumatoide (AR). No obstante, la etnicidad es un factor clave a considerar en desórdenes autoinmunes. Objetivos: Comparar las alteraciones fenotípicas y funcionales de las CDs obtenidas desde pacientes Chilenos con AR y controles sanos (CS). Además, analizamos las diferencias en el comportamiento inflamatorio que existe entre las CDs obtenidas de pacientes fumadores y CS. Materiales y Métodos: Se obtuvieron CDs derivadas de monocitos y células T desde muestras de sangre aisladas de 30 CS y 32 pacientes con AR, 14 de los cuales eran fumadores y 18 no fumadores. Se evaluaron marcadores de maduración en la superficie de las CDs: HLA-DR, CD40, CD80, CD83 y CD86. Además, se realizaron co-cultivos autólogos de células T y CDs, analizando la proliferación de células T, y la expansión de células Th1, Th17 y Tregs. Resultados: En comparación con los CS, los pacientes AR mostraron un aumento de la expresión de HLA-DR en las CDs, principalmente en los individuos con DAS28 moderado-alto. Los pacientes con AR presentaron una mayor expansión de células Th17 y una correlación entre el DAS28 y la expansión de células Th1, ambos efectos manifestados principalmente en los individuos con un DAS28 bajo o en remisión. Además, los pacientes con AR fumadores mostraron un aumento en la expresión de HLA-DR y CD83 en las CDs y una expansión de células Th17 exacerbada así como una correlación entre el DAS28 y la expansión de células Th1. Conclusiones: Nuestros resultados sugieren que fumar favorece el comportamiento inflamatorio de las CDs y en consecuencia la inducción de respuestas mediadas por células Th1 y Th17 en los pacientes Chilenos con AR.


Subject(s)
Humans , Female , Middle Aged , Arthritis, Rheumatoid/metabolism , Dendritic Cells/immunology , Smoking/adverse effects , Cell Proliferation/physiology , Phenotype , Arthritis, Rheumatoid/physiopathology , Arthritis, Rheumatoid/immunology , Smoking/physiopathology , Antigens, Differentiation, B-Lymphocyte/immunology , HLA-DR Antigens/immunology , Case-Control Studies , Chile , T-Lymphocyte Subsets/immunology , Disease Progression , Flow Cytometry , Inflammation/physiopathology , Inflammation/drug therapy
3.
Arq. neuropsiquiatr ; 74(1): 5-9, Jan. 2016. graf
Article in English | LILACS | ID: lil-772601

ABSTRACT

The mechanisms involved in the symptoms of Sydenham’s chorea (SC) remain obscure. Taking into account the autoreactive antibody-mediated hypothesis of SC pathogenesis, the persistence of chorea may be associated with increased levels of B1 lymphocytes and other lymphocyte subsets. We evaluated lymphocyte subsets, including B1 and T cells, in patients with remitted (RSC) and persistent (PSC) SC by flow cytometry. Our results showed neither difference in the frequency of T and B lymphocytes subpopulations nor in their activation and functional states. These findings undermine the view of PSC as a sustained cytotoxic cellular-mediated condition. Alternative mechanisms may explain the pathogenesis of PSC.


Os mecanismos subjacentes aos sintomas da coreia de Sydenham (CS) permanecem desconhecidos. Considerando-se a hipótese de que a patogênese da CS é mediada por anticorpos autorreativos, a persistência da coreia está provavelmente associada a níveis aumentados de linfócitos B1 e outros subtipos de linfócitos. No presente trabalho, foram avaliados subtipos de linfócitos B e T em pacientes com CS em remissão (CSR) e persistente (CSP), por citometria de fluxo. Nossos resultados demonstraram que não há diferença na frequência das subpopulações de linfócitos T e B circulantes e no perfil de ativação e estado funcional dessas células. Esses resultados enfraquecem a hipótese de que a CSP seja uma condição imune sustentada mediada por células citotóxicas. São necessários estudos que investiguem mecanismos alternativos que expliquem a patogênese da CSP.


Subject(s)
Adolescent , Adult , Female , Humans , Male , Young Adult , Autoimmunity/physiology , B-Lymphocyte Subsets/pathology , Chorea/immunology , T-Lymphocyte Subsets/pathology , B-Lymphocyte Subsets/immunology , Flow Cytometry , Lymphocyte Count , T-Lymphocyte Subsets/immunology
4.
Braz. j. med. biol. res ; 49(4): e5062, 2016. tab, graf
Article in English | LILACS | ID: biblio-951667

ABSTRACT

Type 2 diabetes mellitus (T2D) is a metabolic disease with inflammation as an important pathogenic background. However, the pattern of immune cell subsets and the cytokine profile associated with development of T2D are unclear. The objective of this study was to evaluate different components of the immune system in T2D patients' peripheral blood by quantifying the frequency of lymphocyte subsets and intracellular pro- and anti-inflammatory cytokine production by T cells. Clinical data and blood samples were collected from 22 men (51.6±6.3 years old) with T2D and 20 nonsmoking men (49.4±7.6 years old) who were matched for age and sex as control subjects. Glycated hemoglobin, high-sensitivity C-reactive protein concentrations, and the lipid profile were measured by a commercially available automated system. Frequencies of lymphocyte subsets in peripheral blood and intracellular production of interleukin (IL)-4, IL-10, IL-17, tumor necrosis factor-α, and interferon-γ cytokines by CD3+ T cells were assessed by flow cytometry. No differences were observed in the frequency of CD19+ B cells, CD3+CD8+ and CD3+CD4+ T cells, CD16+56+ NK cells, and CD4+CD25+Foxp3+ T regulatory cells in patients with T2D compared with controls. The numbers of IL-10- and IL-17-producing CD3+ T cells were significantly higher in patients with T2D than in controls (P<0.05). The frequency of interferon-γ-producing CD3+ T cells was positively correlated with body mass index (r=0.59; P=0.01). In conclusion, this study shows increased numbers of circulating IL-10- and IL-17-producing CD3+ T cells in patients with T2D, suggesting that these cytokines are involved in the immune pathology of this disease.


Subject(s)
Humans , Male , Adult , Middle Aged , Cytokines/blood , T-Lymphocyte Subsets/metabolism , Diabetes Mellitus, Type 2/blood , Reference Values , C-Reactive Protein/metabolism , T-Lymphocytes/cytology , T-Lymphocytes/immunology , T-Lymphocytes/metabolism , Case-Control Studies , T-Lymphocyte Subsets/cytology , T-Lymphocyte Subsets/immunology , Statistics, Nonparametric , Lymphocyte Count , Diabetes Mellitus, Type 2/immunology , Flow Cytometry , Immunity, Cellular
5.
Braz. j. med. biol. res ; 48(12): 1095-1100, Dec. 2015. graf
Article in English | LILACS | ID: lil-762920

ABSTRACT

In DNA vaccines, the gene of interest is cloned into a bacterial plasmid that is engineered to induce protein production for long periods in eukaryotic cells. Previous research has shown that the intramuscular immunization of BALB/c mice with a naked plasmid DNA fragment encoding the Mycobacterium leprae 65-kDa heat-shock protein (pcDNA3-Hsp65) induces protection against M. tuberculosis challenge. A key stage in the protective immune response after immunization is the generation of memory T cells. Previously, we have shown that B cells capture plasmid DNA-Hsp65 and thereby modulate the formation of CD8+ memory T cells after M. tuberculosis challenge in mice. Therefore, clarifying how B cells act as part of the protective immune response after DNA immunization is important for the development of more-effective vaccines. The aim of this study was to investigate the mechanisms by which B cells modulate memory T cells after DNA-Hsp65 immunization. C57BL/6 and BKO mice were injected three times, at 15-day intervals, with 100 µg naked pcDNA-Hsp65 per mouse. Thirty days after immunization, the percentages of effector memory T (TEM) cells (CD4+ and CD8+/CD44high/CD62Llow) and memory CD8+ T cells (CD8+/CD44high/CD62Llow/CD127+) were measured with flow cytometry. Interferon γ, interleukin 12 (IL-12), and IL-10 mRNAs were also quantified in whole spleen cells and purified B cells (CD43−) with real-time qPCR. Our data suggest that a B-cell subpopulation expressing IL-10 downregulated proinflammatory cytokine expression in the spleen, increasing the survival of CD4+ TEM cells and CD8+ TEM/CD127+ cells.


Subject(s)
Animals , Male , Mice , B-Lymphocytes/immunology , Heat-Shock Proteins/immunology , Immunomodulation/genetics , /genetics , RNA, Messenger/immunology , T-Lymphocyte Subsets/immunology , B-Lymphocytes/metabolism , Flow Cytometry , Gene Expression/genetics , Heat-Shock Proteins/therapeutic use , Immunologic Memory/physiology , Immunophenotyping/classification , Inflammation Mediators/analysis , Interferon-gamma/analysis , /immunology , /analysis , Mice, Knockout , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , RNA, Messenger/genetics , Spleen/cytology , Spleen/immunology , T-Lymphocyte Subsets/classification , Vaccines, DNA/immunology , Vaccines, DNA/therapeutic use
6.
Rev. Assoc. Med. Bras. (1992) ; 61(4): 329-335, July-Aug. 2015. tab, ilus
Article in English | LILACS | ID: lil-761708

ABSTRACT

SummaryIntroduction:aging is associated with several immunologic changes. Regulatory (Treg) and effector T cells are involved in the pathogenesis of infectious, neoplastic, and autoimmune diseases. Little is known about the effects of aging on the frequency and function of these T cell subpopulations.Methods:peripheral blood mononuclear cells (PBMC) were obtained from 26 young (under 44 years old) and 18 elderly (above 80 years old) healthy women. T cell subpopulations were analyzed by flow cytometry.Results:elderly individuals had lower frequency of several activated effector T cell phenotypes as compared with young individuals: CD3+CD4+CD25+ (3.82±1.93 versus 9.53±4.49; p<0.0001); CD3+CD4+CD25+CD127+(2.39±1.19 versus 7.26±3.84; p<0.0001); CD3+CD4+CD25+ (0.41±0.22 versus 1.86±0.85, p<0.0001); and CD3+CD4+CD25highCD127+(0.06±0.038 versus 0.94±0.64, p<0.0001). Treg (CD3+CD4+CD25+CD127øFoxp3+) presented lower frequency in elderly individuals as compared to young adults (0.34±0.18 versus 0.76±0.48; p=0.0004) and its frequency was inversely correlated with age in the whole group (r=-0.439; p=0.013). The elderly group showed higher frequency of two undefined CD25øFoxp3+ phenotypes: CD3+CD4+CD25øFoxp3+(15.05±7.34 versus 1.65±1.71; p<0.0001) and CD3+CD4+CD25øCD127øFoxp3+(13.0±5.52 versus 3.51±2.87; p<0.0001).Conclusions:the altered proportion of different T cell subsets herein documented in healthy elderly women may be relevant to the understanding of the immunologic behavior and disease susceptibility patterns observed in geriatric patients.


ResumoIntrodução:o envelhecimento está associado a diversas alterações imunológicas. Células T reguladoras e efetoras estão envolvidas na patogênese de enfermidades infecciosas, neoplásicas e autoimunes. Pouco se sabe acerca dos efeitos da idade sobre a frequência e a função dessas populações celulares.Métodos:células mononucleares do sangue periférico foram obtidas de participantes saudáveis (26 com idade inferior a 44 anos e 18 acima de 80 anos). As subpopulações celulares foram analisadas por citometria de fluxo.Resultados:o grupo constituído por idosas apresentou menor frequência de vários fenótipos de células T efetoras ativadas em comparação com jovens: CD3+CD4+CD25+ (3,82±1,93 versus 9,53±4,49, p<0,0001); CD3+CD4+ CD25+CD127+ (2,39±1,19 versus7,26±3,84, p<0,0001); CD3+CD4+CD25high(0,41±0,22 versus 1,86±0,85, p<0,0001); CD3+CD4+CD25highCD127+(0,06±0,038 versus 0,94±0,64, p<0,0001). As células T reguladoras CD3+CD4+CD25highCD127øFoxP3+ apresentaram menor frequência em indivíduos idosos em comparação com adultos jovens (0,34±0,18 versus0,76±0,48, p=0,0004) e sua frequência foi inversamente correlacionada com a idade em todo o grupo (r=-0,439; p=0,013). O grupo de idosas apresentou maior frequência de dois fenótipos indefinidos (CD25øFoxP3+), células CD3+CD4+CD25øFoxP3+ (15,05±7,34 versus 1,65±1,71, p<0,0001) e células CD3+CD4+CD25øCD127øFoxP3+(13,0±5,52 versus 3,51±2,87, p<0,0001).Conclusão:as proporções alteradas de diferentes subpopulações de células T em idosas saudáveis contribuem para a compreensão dos padrões de comportamento e suscetibilidade a doenças imunológicas evidenciadas em pacientes geriátricos.


Subject(s)
Adult , Aged, 80 and over , Female , Humans , Young Adult , Aging/immunology , Immunophenotyping , T-Lymphocyte Subsets/immunology , T-Lymphocytes, Regulatory/cytology , Age Factors , Flow Cytometry , Leukocytes, Mononuclear/cytology , Leukocytes, Mononuclear/immunology , T-Lymphocyte Subsets/cytology , T-Lymphocytes, Regulatory/immunology
7.
Mem. Inst. Oswaldo Cruz ; 110(5): 596-605, Aug. 2015. tab, ilus
Article in English | LILACS | ID: lil-755895

ABSTRACT

In human cutaneous leishmaniasis (CL), the immune response is mainly mediated by T-cells. The role of CD8+ T-lymphocytes, which are related to healing or deleterious functions, in affecting clinical outcome is controversial. The aim of this study was to evaluate T-cell receptor diversity in late-differentiated effector (LDE) and memory CD8+ T-cell subsets in order to create a profile of specific clones engaged in deleterious or protective CL immune responses. Healthy subjects, patients with active disease (PAD) and clinically cured patients were enrolled in the study. Total CD8+ T-lymphocytes showed a disturbance in the expression of the Vβ2, Vβ9, Vβ13.2, Vβ18 and Vβ23 families. The analyses of CD8+T-lymphocyte subsets showed high frequencies of LDE CD8+T-lymphocytes expressing Vβ12 and Vβ22 in PAD, as well as effector-memory CD8+ T-cells expressing Vβ22. We also observed low frequencies of effector and central-memory CD8+ T-cells expressing Vβ2 in PAD, which correlated with a greater lesion size. Particular Vβ expansions point to CD8+ T-cell clones that are selected during CL immune responses, suggesting that CD8+ T-lymphocytes expressing Vβ12 or Vβ22 are involved in a LDE response and that Vβ2 contractions in memory CD8+T-cells are associated with larger lesions.

.


Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Young Adult , /immunology , Leishmaniasis, Cutaneous/immunology , Receptors, Antigen, T-Cell/immunology , T-Lymphocyte Subsets/immunology , Brazil , Lymphocyte Activation/immunology , Receptors, Antigen, T-Cell/analysis
8.
Article in English | IMSEAR | ID: sea-158451

ABSTRACT

Background & objectives: Since the 2006 massive outbreaks, chikungunya (CHIK) is a major public health concern in India. The aim of this study was to assess envelope specific immune responses in patients with chikungunya infection. Methods: This study included 46 hospitalized patients with chikungunya virus infection (encephalitis, n=22, other systemic involvement, OSI, n=12, classical, n=12) and six controls from Ahmedabad city, Gujarat, India. T cell responses and the levels of Th1, pro/ anti-inflammatory cytokines against the CHIK virus envelope antigens were assessed by lymphocyte proliferation assay and by cytometric bead array in flow cytometry, respectively. Results: Lymphoproliferative response was uniform among the patients. Comparisons of cytokines revealed significantly higher levels of interleukin (IL)-4 and IL-5 in encephalitis, OSI and classical patients versus controls. The levels of tumour necrosis factor (TNF)-α were higher in classical patients categories compared to the controls. Interferon (IFN)-γ levels were lower in encephalitis patients versus control. Interpretation & conclusions: Our findings showed recognition of T cell epitopes on the envelope region of chikungunya virus by all patient categories. Lower level of IFN-γ may be associated with the severity of disease in these patients.


Subject(s)
Chikungunya Fever/diagnosis , Chikungunya Fever/epidemiology , Chikungunya Fever/immunology , Cytokines/analysis , Humans , India , T-Lymphocytes/immunology , T-Lymphocyte Subsets/immunology
9.
Rev. Esc. Enferm. USP ; 48(spe): 102-108, 08/2014. tab
Article in English | LILACS, BDENF | ID: lil-731295

ABSTRACT

Exploratory and descriptive study based on quantitative and qualitative methods that analyze the phenomenon of violence against adolescents based on gender and generational categories. The data source was reports of violence from the Curitiba Protection Network from 2010 to 2012 and semi-structured interviews with 16 sheltered adolescents. Quantitative data were analyzed using SPSS software version 20.0 and the qualitative data were subjected to content analysis. The adolescents were victims of violence in the household and outside of the family environment, as victims or viewers of violence. The violence was experienced at home, mostly toward girls, with marked overtones of gender violence. More than indicating the magnitude of the issue, this study can give information to help qualify the assistance given to victimized people and address how to face this issue.


Objetivo Analizar la violencia contra los adolescentes a la luz de las categorías de género y generación. Método Estudio exploratorio, descriptivo, de abordaje cuantitativo y cualitativo que. Las fuentes de datos fueron las denuncias de violencia mantenidos por la Red de Protección en Curitiba entre los años 2010-2012 y entrevistas semi-estructuradas con 16 adolescentes alojados. Las variables cuantitativas se analizaron mediante el programa SPSS y los cualitativos por la análisis de contenido. Resultados Los adolescentes fueron sometidos a la violencia en el hogar y en el exterior, como víctimas o espectadores. La violencia fue más frecuente en el hogar, centrándose principalmente en las chicas con matices marcados de violencia de género. Conclusión Más que encontrar la magnitud del problema, el estudio puede servir de base para calificar la asistencia a las personas víctimas de este fenómeno.

 .


Objetivo Analisar a violência contra o adolescente à luz das categorias gênero e geração. Método Estudo exploratório, descritivo, de abordagem quantitativa e qualitativa. As fontes de dados foram as notificações de violência da Rede de Proteção do município de Curitiba, de 2010 a 2012, e entrevistas semiestruturadas com 16 adolescentes abrigados. As variáveis quantitativas foram analisadas pelo software SPSS e os dados qualitativos através da análise de conteúdo. Resultados Os adolescentes foram submetidos à violência no ambiente doméstico e fora dele, como vítimas ou como espectadores. Prevaleceu no domicílio, incidindo principalmente sobre as meninas, com marcada conotação de violência de gênero. Conclusão Mais que constatar a magnitude do problema, o estudo pode fornecer subsídios para qualificar a assistência prestada aos sujeitos vitimizados e subsidiar o enfrentamento do fenômeno. .


Subject(s)
Adult , Humans , Middle Aged , Stomach Neoplasms/genetics , Stomach Neoplasms/immunology , Telomerase/genetics , DNA-Binding Proteins , Gene Expression , Immunity, Cellular , Killer Cells, Natural/immunology , Precancerous Conditions/enzymology , Precancerous Conditions/genetics , Precancerous Conditions/immunology , RNA, Messenger/genetics , RNA, Neoplasm/genetics , Stomach Neoplasms/enzymology , T-Lymphocyte Subsets/immunology
10.
J. appl. oral sci ; 22(4): 336-346, Jul-Aug/2014. tab, graf
Article in English | LILACS | ID: lil-718287

ABSTRACT

Previous studies demonstrate that the balance between pro- and anti-inflammatory mediators determines the stable or progressive nature of periapical granulomas by modulating the balance of the osteoclastogenic factor RANKL and its antagonist OPG. However, the cytokine networks operating in the development of periapical lesions are quite more complex than what the simple pro- versus anti-inflammatory mediators' paradigm suggests. Here we simultaneously investigated the patterns of Th1, Th2, Th9, Th17, Th22, Thf, Tr1 and Tregs cytokines/markers expression in human periapical granulomas. Methods: The expression of TNF-α, IFN-γ, IL-17A, IL23, IL21, IL-33, IL-10, IL-4, IL-9, IL-22, FOXp3 markers (via RealTimePCR array) was accessed in active/progressive (N=40) versus inactive/stable (N=70) periapical granulomas (as determined by RANKL/OPG expression ratio), and also to compare these samples with a panel of control specimens (N=26). A cluster analysis of 13 cytokine levels was performed to examine possible clustering between the cytokines in a total of 110 granulomas. Results: The expression of all target cytokines was higher in the granulomas than in control samples. TNF-α, IFN-γ, IL-17A and IL-21 mRNA levels were significantly higher in active granulomas, while in inactive lesions the expression levels of IL-4, IL-9, IL-10, IL-22 and FOXp3 were higher than in active granulomas. Five clusters were identified in inactive lesion groups, being the variance in the expression levels of IL-17, IL-10, FOXp3, IFN-γ, IL-9, IL-33 and IL-4 statistically significant (KW p<0.05). Three clusters were identified in active lesions, being the variance in the expression levels of IL-22, IL-10, IFN-γ, IL-17, IL-33, FOXp3, IL-21 and RANKL statistically significant (KW p<0.05). Conclusion: There is a clear dichotomy in the profile of cytokine expression in inactive and active periapical lesions. While the widespread ...


Subject(s)
Adult , Female , Humans , Male , Middle Aged , Young Adult , Cytokines/analysis , Periapical Granuloma/pathology , T-Lymphocyte Subsets/metabolism , T-Lymphocytes, Helper-Inducer/metabolism , Analysis of Variance , Biomarkers/analysis , Chronic Disease , Cytokines/immunology , Periapical Granuloma/immunology , Real-Time Polymerase Chain Reaction , Reference Values , Statistics, Nonparametric , T-Lymphocyte Subsets/immunology , T-Lymphocytes, Helper-Inducer/immunology
11.
Article in English | WPRIM | ID: wpr-80828

ABSTRACT

BACKGROUND: We developed a single-color multitarget flow cytometry (SM-FC) assay, a single-tube assay with graded mean fluorescence intensities (MFIs). We evaluated the repeatability of SM-FC, and its correlation with multicolor flow cytometry (MFC), to assess its application as a routine FC assay. METHODS: We selected CD19, CD3, CD4, and CD8 as antigen targets to analyze a lymphocyte subset. MFIs were graded by adjusting monoclonal antibody (mAb) volumes to detect several cell populations. Dimly labeled mAb was prepared by decreasing mAb volume and the optimum diluted volume was determined by serial dilution. SM-FC repeatability was analyzed 10 times in 2 normal controls. The correlation between SM-FC and MFC was evaluated in 20 normal and 23 patient samples. RESULTS: CV values (0.8-5.0% and 1.3-4.1% in samples 1 and 2, respectively) acquired by SM-FC with CD3-fluorescein alpha-isothyocyanate (FITC)dim+CD4-FITCbright and with CD19-FITCdim+CD3-FITCbright showed good repeatability, comparable to that acquired by MFC (1.6-3.7% and 1.0-4.8% in samples 1 and 2, respectively). Excellent correlation was observed between the 2 methods in the 20 normal samples (B cells, T cells, non-Thelper cells, and Thelper cells; r2=0.87, 0.97, 0.97, and 0.98, respectively; P or =0.98, 0.99, 0.99, and 0.99, respectively; P<0.05). CONCLUSIONS: The multicolor, single-tube SM-FC technique is a potential alternative tool for identifying a lymphocyte subset.


Subject(s)
Antibodies, Monoclonal/chemistry , Antigens, CD19/chemistry , CD3 Complex/chemistry , CD4 Antigens/chemistry , CD8 Antigens/chemistry , B-Lymphocyte Subsets/immunology , Color , Flow Cytometry/methods , Fluorescein-5-isothiocyanate/chemistry , Humans , T-Lymphocyte Subsets/immunology
12.
Invest. clín ; 52(4): 365-375, dic. 2011. ilus
Article in English | LILACS | ID: lil-659226

ABSTRACT

A patient with localized cutaneous leishmaniasis due to Leishmania (Leishmania) amazonensis infection was treated with an antigen containing heat-killed L. (L.) amazonensis promastigotes plus BCG. Expression of T-cell differentiation, memory and senescence receptors markers were analyzed on T cell subpopulations, in order to establish the correlation between the percentages of expression of these receptors and his clinical status, at different stages of his follow up. The following case reports on the achievement of a successful clinical outcome with complete resolution after receiving immunotherapy. A thorough clinical and immunological follow up supporting the healing process of this patient’s lesion is presented in detail.


Un paciente con leishmaniasis cutánea localizada producida por Leishmania (Leishmania) amazonensis fue tratado con un antígeno compuesto por promastigotes de L. (L.) amazonensis muertos por calor combinado con BCG. Se analizó la expresión de distintos receptores de diferenciación, de memoria y de senescencia en las subpoblaciones de células T, con el fin de establecer una relación entre los porcentajes de expresión de dichos receptores y la clínica del paciente en diferentes momentos del seguimiento. Se reporta en este caso un resultado exitoso, con resolución completa de la lesión después de recibir la inmunoterapia, y se presenta en detalle un seguimiento clínico e inmunológico completo durante el proceso de curación.


Subject(s)
Adult , Humans , Male , Antigens, Protozoan/therapeutic use , BCG Vaccine/therapeutic use , Immunotherapy, Active , Leishmania mexicana/immunology , Leishmaniasis, Cutaneous/therapy , Occupational Diseases/therapy , Protozoan Vaccines/therapeutic use , Antigens, Protozoan/administration & dosage , Antigens, Protozoan/immunology , Argentina/epidemiology , BCG Vaccine/administration & dosage , Fisheries , Immunity, Cellular , Immunologic Memory , Injections, Intradermal , Leg Ulcer/etiology , Leg Ulcer/parasitology , Leishmania mexicana/growth & development , Leishmaniasis, Cutaneous/epidemiology , Leishmaniasis, Cutaneous/immunology , Leishmaniasis, Cutaneous/parasitology , Occupational Diseases/immunology , Occupational Diseases/parasitology , Protozoan Vaccines/administration & dosage , Protozoan Vaccines/immunology , T-Lymphocyte Subsets/immunology , Vaccines, Inactivated
13.
Article in English | IMSEAR | ID: sea-137359

ABSTRACT

Background & objectives: HIV infection is characterized by a perturbation in T cell homeostasis, leading to alteration in T cell subsets. In addition to alteration in differentiation, HIV infection also leads to change in T cell survival and regenerative capacity, as suggested by differential expression of CD127 and CD57. We evaluated the expression patterns of CD127 and CD57 on CD4 and CD8 effector, memory and naïve T cell subsets in HIV-infected and uninfected individuals. Methods: We characterized T cell subsets based on expression of these markers, and compared their expression pattern in HIV infected subjects and uninfected controls. We further assessed therapy generated changes in these subsets and expression of CD127 and CD57 on them. Results: There was a generalized decrease in naïve CD4 and CD8 T cells in HIV infected subjects. These changes in T cell subset distribution were related to antigen load. CD127 expression was significantly reduced in T cells from HIV infected subject. In association to this, HIV infected subjects had higher percentage of T cell subsets expressing CD57. Increased CD57 and reduced CD127 expression correlated with plasma viraemia and CD8 T cell activation state. Incomplete restoration of T cell subset proportions was observed, despite suppression of viral replication and increase in CD4 T cell counts. Further, the improvement was more pronounced in CD127 expression. Interpretation & conclusions: HIV infected subjects have reduced T cell regenerative capacity along with increased senescence, highlighting decreased proliferation and effector activities.


Subject(s)
Adult , CD57 Antigens/metabolism , Antiretroviral Therapy, Highly Active , CD4 Lymphocyte Count , CD4-CD8 Ratio , Cell Differentiation/immunology , Female , HIV Infections/drug therapy , Humans , HIV Infections/immunology , Immunophenotyping , Interleukin-7 Receptor alpha Subunit/deficiency , Interleukin-7 Receptor alpha Subunit/metabolism , Male , Statistics, Nonparametric , T-Lymphocyte Subsets/immunology
14.
Article in English | WPRIM | ID: wpr-82228

ABSTRACT

Natural killer (NK) cells play an important role in innate immunity, especially in the response to viral infections, such as hepatitis C virus (HCV). Killer cell immunoglobulin-like receptors (KIRs) are the primary receptors of NK cells that mediate innate immunity. KIRs are also involved in acquired immunity, because some KIRs are expressed on the surface of certain subsets of T cells. In this study, the frequency of KIR genes, HLA-C allotypes, and combinations of KIR genes with their HLA-C ligands were evaluated in two different groups of the Korean population: controls and patients with chronic HCV infection. The study population consisted of 147 Korean patients with chronic HCV infection. The frequency of KIR2DS2 in patients with chronic HCV infection was 9.5% which was significantly lower than 19.5% of the control (P < 0.01). However, there were no significant differences in the frequency of other KIR genes, HLA-C allotypes or different combinations of KIR genes with their HLA-C ligands. This study can contribute to the further prospective study with a larger scale, suggesting the assumption that KIR2DS2 might aid in HCV clearance by enhancing both the innate and acquired immune responses of people in Korea.


Subject(s)
Adult , Aged , Female , Genes, MHC Class I , Genotype , HLA-C Antigens/genetics , Hepacivirus/immunology , Hepatitis C, Chronic/genetics , Humans , Killer Cells, Natural/immunology , Male , Middle Aged , Receptors, KIR/genetics , Republic of Korea , T-Lymphocyte Subsets/immunology
15.
Article in English | WPRIM | ID: wpr-47951

ABSTRACT

Toxoplasma gondii Korean isolate (KI-1) tachyzoites were inoculated intraduodenally to BALB/c mice using a silicon tube, and the course of infection and immune responses of mice were studied. Whereas control mice, that were infected intraperitoneally, died within day 7 post-infection (PI), the intraduodenally infected mice survived until day 9 PI (infection with 1x10(5) tachyzoites) or day 11 PI (with 1x10(6) tachyzoites). Based on histopathologic (Giemsa stain) and PCR (B1 gene) studies, it was suggested that tachyzoites, after entering the small intestine, invaded into endothelial cells, divided there, and propagated to other organs. PCR appeared to be more sensitive than histopathology to detect infected organs and tissues. The organisms spread over multiple organs by day 6 PI. However, proliferative responses of splenocytes and mesenteric lymph node (MLN) cells in response to con A or Toxoplasma lysate antigen decreased significantly, suggesting immunosuppression. Splenic CD4+ and CD8+ T-lymphocytes showed decreases in number until day 9 PI, whereas IFN-gamma and IL-10 decreased slightly at day 6 PI and returned to normal levels by day 9 PI. No TNF-alpha was detected throughout the experimental period. The results showed that intraduodenal infection with KI-1 tachyzoites was successful but did not elicit significant mucosal immunity in mice and allowed dissemination of T. gondii organisms to systemic organs. The immunosuppression of mice included reduced lymphoproliferative responses to splenocytes and MLN cells to mitogen and low production of cytokines, such as IFN-gamma, TNF-alpha, and IL-10, in response to T. gondii infection.


Subject(s)
Animals , Cell Proliferation , Cytokines/metabolism , Disease Models, Animal , Duodenum/immunology , Endothelial Cells/parasitology , Histocytochemistry , Immune Tolerance , Lymph Nodes/immunology , Mice , Mice, Inbred BALB C , Polymerase Chain Reaction , Rodent Diseases/immunology , T-Lymphocyte Subsets/immunology , Toxoplasma/immunology , Toxoplasmosis, Animal/immunology
16.
Article in English | WPRIM | ID: wpr-199407

ABSTRACT

Most of thyroid lymphomas are B-lineage, and T-cell lymphomas are rare. Here, we report a case of primary thyroid T-cell lymphoma associated with Hashimoto's thyroiditis. A 48-yr-old woman presented with incidentally found neck mass. Histologically, the resected right lobe of the thyroid was replaced by monomorphic small atypical lymphoid cells with lymphoepithelial lesion-like change, most of which were immunoreactive for CD3, CD8, betaF-1, and TIA-1. Peripheral T-cell lymphoma, unspecified, was finally diagnosed after molecular study for TCR-gamma gene rearrangement. This is the second case of cytotoxic T-cell lymphoma reported in the thyroid gland so far. Unique association between thyroid follicles and neoplastic lymphocytes may be characteristic feature of this type of T-cell lymphoma.


Subject(s)
Female , Hashimoto Disease/pathology , Humans , Lymphoma, B-Cell, Marginal Zone/pathology , Lymphoma, T-Cell/pathology , Middle Aged , T-Lymphocyte Subsets/immunology , T-Lymphocytes, Cytotoxic/immunology , Thyroid Gland/pathology
17.
Braz. j. med. biol. res ; 42(6): 476-486, June 2009. ilus, tab
Article in English | LILACS | ID: lil-512764

ABSTRACT

A new subtype of CD4+ T lymphocytes characterized by the production of interleukin 17, i.e., TH17 cells, has been recently described. This novel T cell subset is distinct from type 1 and type 2 T helper cells. The major feature of this subpopulation is to generate significant amounts of pro-inflammatory cytokines, therefore appearing to be critically involved in protection against infection caused by extracellular microorganisms, and in the pathogenesis of autoimmune diseases and allergy. The dynamic balance among subsets of T cells is important for the modulation of several steps of the immune response. Disturbances in this balance may cause a shift from normal immunologic physiology to the development of immune-mediated disorders. In autoimmune diseases, the fine balance between the proportion and degree of activation of the various T lymphocyte subsets can contribute to persistent undesirable inflammatory responses and tissue replacement by fibrosis. This review highlights the importance of TH17 cells in this process by providing an update on the biology of these cells and focusing on their biology and differentiation processes in the context of immune-mediated chronic inflammatory diseases.


Subject(s)
Animals , Humans , Mice , Autoimmune Diseases/immunology , /immunology , /biosynthesis , T-Lymphocyte Subsets/immunology , Disease Models, Animal , T-Lymphocyte Subsets
18.
Indian J Med Microbiol ; 2009 Jan-Mar; 27(1): 40-3
Article in English | IMSEAR | ID: sea-53930

ABSTRACT

T cells have the capability of recognizing target cells through their T cell receptors (TCRs). Thus, the percentages of CD3+/gamma-delta (gammadelta) TCR+ and CD3+/alpha-beta (alphabeta) TCR+ T lymphocytes were investigated in active and inactive pulmonary tuberculosis (PT) patients and in healthy individuals. CD3+ and CD3+/alphabeta TCR+ cell percentages were significantly lower in all PT patients than in healthy subjects. Percentages of CD3+/gammadelta and CD3+/alphabeta TCR+ were not statistically different between active and inactive PT patients. It was concluded that alphabeta TCR+ T cells might have a protective role in tuberculosis infection.


Subject(s)
Adult , CD3 Complex/analysis , Blood/immunology , Female , Humans , Male , Middle Aged , Receptors, Antigen, T-Cell/immunology , T-Lymphocyte Subsets/immunology , T-Lymphocytes/immunology , Tuberculosis, Pulmonary/immunology , Young Adult
19.
Article in English | WPRIM | ID: wpr-71512

ABSTRACT

In addition to CD4+CD25+Foxp3+ regulatory T (T(reg)) cells which protect against autoimmune tissue injury, IL-17-producing CD4+ T (Th17) cells have been recently described and shown to play a crucial role in autoimmune injury. It appears that there is a reciprocal developmental pathway between Th17 and T(reg) cells. Although IL-17 is known to be associated with allograft rejection, the cellular source of IL-17 and the nature of Th17 in the context of allograft rejection remain unknown. In the current study, the dynamics of T(reg) and IL-17-producing cells after syngeneic and allogeneic transplantation were examined using a wild-type murine cardiac transplantation model. Ly6G+ cells were found to produce IL-17 during the early postoperative period and CD8+ as well as CD4+ T cells were also found to produce IL-17 during alloimmune response. Graft-infiltrating Ly6G+, CD4+, and even CD8+ cells were found to express IL-17 highly compared to those in spleen. Although the frequencies of Th17 and T(reg) were found to gradually increase in both syngeneic and allogeneic recipients, Th17/T(reg) ratios were significantly higher in recipients with allograft rejection than in syngeneic recipients. In conclusion, IL-17 is produced by neutrophils during the early postoperative period and subsequently by Th17 and CD8+ T cells during allograft rejection. Th17/T(reg) imbalance is associated with the development of allograft rejection. This study would provide basic information on Th17 biology for future investigation in the field of transplantation.


Subject(s)
Animals , Antigens, CD/biosynthesis , Autoimmunity , Forkhead Transcription Factors/biosynthesis , Graft Rejection/immunology , Heart Transplantation , Interleukin-17/immunology , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Neutrophils/immunology , T-Lymphocyte Subsets/immunology , T-Lymphocytes, Regulatory/immunology , Time Factors , Transplantation Immunology
20.
Clinics ; 63(5): 637-644, 2008. tab
Article in English | LILACS | ID: lil-495039

ABSTRACT

INTRODUCTION AND OBJECTIVES: Tuberculosis and cancer are the main causes of pleural effusion. Pleural involvement is associated with migration of immune cells to the pleural cavity. We sought to characterize the immunophenotype of leukocytes in the pleural effusion and peripheral blood of patients with tuberculosis or malignancy. METHODS: Thirty patients with tuberculosis (14) or malignancy (16) were studied. A control group included 20 healthy blood donors. RESULTS: Malignant phycoerythrin pleural effusions showed higher percentages of CD3, CD4, CD3CD45RO, and CD20CD25 lymphocytes and lower percentages of CD3CD25 and CD20HLA-DR when compared to PB lymphocytes. Compared to PB, tuberculous effusions had a higher percentage of lymphocytes that co-expressed CD3, CD4, CD3CD45RO, CD3TCRáâ, CD3CD28, and CD20 and a lower percentage of CD14, CD8 and CD3TCRãä-positive lymphocytes. Malignant effusions presented higher expression of CD14 whereas tuberculous effusions had higher expression of CD3 and CD3CD95L. Peripheral blood cells from tuberculosis patients showed higher expression of CD14, CD20CD25 and CD3CD95L. Compared with the control cells, tuberculosis and cancer peripheral blood cells presented a lower percentage of CD3CD4 and CD3CD28-positive cells as well as a higher percentage of CD3CD8, CD3CD25 and CD3CD80-positive cells. CONCLUSIONS: Tuberculous and malignant peripheral blood is enriched with lymphocytes with a helper/inducer T cell phenotype, which are mainly of memory cells. CD14-positive cells were more frequently found in malignant effusions, while CD3-positive cells expressing Fas ligand were more frequently found in tuberculous effusions.


Subject(s)
Adult , Female , Humans , Male , Middle Aged , /immunology , Immunophenotyping , Pleural Effusion, Malignant/immunology , T-Lymphocyte Subsets/immunology , Tuberculosis, Pleural/immunology , Analysis of Variance , Apoptosis , Case-Control Studies , Exudates and Transudates/immunology , Flow Cytometry , Immunity, Cellular , Pleural Effusion, Malignant/blood , Statistics, Nonparametric , Tuberculosis, Pleural/blood
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