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1.
Adv Exp Med Biol ; 839: 1-5, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25252903

RESUMO

Recent studies show that several Siglec receptors, such as Siglec-8 and Siglec-14, may be important therapeutic targets in asthma and COPD. Siglecs are a family of lectins belonging to the immunoglobulin superfamily and recognize sialic acid residues of glycoproteins. Most of Siglecs have intracellular immunoreceptor tyrosine-based inhibitory motifs (ITIM), implicating them in the suppression of immunoreceptor signaling. Siglec-5/14 may be involved in the negative regulation of innate immune responses. The aim of this study was to analyze Siglec-5/14 expression in induced sputum cells of COPD patients in the following treatment combinations: (1) a long-acting beta2-agonist, formoterol; (2) formoterol combined with a long-acting antimuscarinic agent, tiotropium; and (3) formoterol combined with an inhaled corticosteroid or formoterol combined with tiotropium and with an inhaled corticosteroid. Siglec expression was assessed in sputum cells by flow cytometry using a specific monoclonal antibody. Double staining of cells indicated that Siglec-5/14 is expressed in monocyte/macrophages and neutrophils, but not in lymphocytes. Siglec-5/14 expression was significantly higher in patients receiving combined therapy including inhaled corticosteroids compared with patients taking only formoterol or formoterol + tiotropium. Our results suggest that inhaled corticosteroids may exert beneficial or negative effects, depending on the patients' phenotype, through increased immunosuppressive Siglec-5 or immunoactivatory Siglec-14 receptors, respectively.


Assuntos
Corticosteroides/uso terapêutico , Antígenos CD/genética , Antígenos de Diferenciação Mielomonocítica/genética , Broncodilatadores/uso terapêutico , Etanolaminas/uso terapêutico , Lectinas/genética , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Receptores de Superfície Celular/genética , Derivados da Escopolamina/uso terapêutico , Agonistas de Receptores Adrenérgicos beta 2/uso terapêutico , Androstadienos/uso terapêutico , Budesonida/uso terapêutico , Separação Celular , Antagonistas Colinérgicos/uso terapêutico , Quimioterapia Combinada , Fluticasona , Fumarato de Formoterol , Expressão Gênica , Humanos , Imunofenotipagem , Lectinas/agonistas , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Macrófagos/patologia , Neutrófilos/efeitos dos fármacos , Neutrófilos/metabolismo , Neutrófilos/patologia , Doença Pulmonar Obstrutiva Crônica/metabolismo , Doença Pulmonar Obstrutiva Crônica/patologia , Receptores de Superfície Celular/agonistas , Escarro/citologia , Escarro/efeitos dos fármacos , Escarro/metabolismo , Linfócitos T/efeitos dos fármacos , Linfócitos T/metabolismo , Linfócitos T/patologia , Brometo de Tiotrópio
2.
Adv Exp Med Biol ; 839: 7-12, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25315616

RESUMO

Immune cells expressing the activation markers HLA-DR and regulatory T cells (Tregs) may be involved in the regulation of chronic inflammation in chronic obstructive pulmonary disease (COPD). In this study we analyzed native and activated cell profiles in sputum of 22 stable COPD patients receiving formoterol (F) or formoterol + tiotropium (F + T) for 3 months. Cells were isolated from induced sputum and were examined on Coulter flow cytometer using fluorescent antibodies specific for CD3, CD4, CD8, CD14, CD19, CD25, CD127, and HLA-DR antigens. Cell profiles and cell activation were assessed by analysis of HLA-DR, CD25, and CD127 co-expression in double-stained samples. Tregs were defined as CD4⁺CD25(high) CD127(low) cells. We found that the combined therapy significantly decreased the CD8⁺ cell number (p < 0.01). At baseline, HLA-DR was expressed in about 10 % of sputum T or B cells and a higher expression was found on monocytes. The HLA-DR expression on lymphocytes, but not monocytes, was significantly lower (p < 0.01) in patients treated with F + T. Fractions of activated [CD4⁺ CD25⁺] cells were also significantly lower in the combined therapy group, except for the subpopulation of CD4⁺CD25(high) CD127(low) cells which was not altered. We conclude that tiotropium in add-on therapy to formoterol affects Treg cell profiles and decreases HLA-DR expression in airway lymphocytes.


Assuntos
Broncodilatadores/uso terapêutico , Etanolaminas/uso terapêutico , Antígenos HLA-DR/genética , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Derivados da Escopolamina/uso terapêutico , Linfócitos T Reguladores/efeitos dos fármacos , Agonistas de Receptores Adrenérgicos beta 2/uso terapêutico , Idoso , Antígenos CD/genética , Separação Celular , Antagonistas Colinérgicos/uso terapêutico , Quimioterapia Combinada , Fumarato de Formoterol , Expressão Gênica , Humanos , Imunofenotipagem , Masculino , Pessoa de Meia-Idade , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , Monócitos/patologia , Doença Pulmonar Obstrutiva Crônica/metabolismo , Doença Pulmonar Obstrutiva Crônica/patologia , Escarro/citologia , Escarro/efeitos dos fármacos , Escarro/metabolismo , Linfócitos T Reguladores/metabolismo , Linfócitos T Reguladores/patologia , Brometo de Tiotrópio
3.
Exp Clin Endocrinol Diabetes ; 120(2): 101-9, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21922455

RESUMO

UNLABELLED: There is increasing evidence that T-regulatory (Treg) cells could be used to prevent or cure autoimmune diseases including type 1 diabetes mellitus (T1DM). The aim of the present study was to verify the hypothesis that functional Treg cells can be generated from conventional T-cells separated from a small amount of peripheral blood of children with newly diagnosed T1DM (N=25). METHODS: CD4(+)CD25(-) cells were cultured with Treg expander (CD3/CD28) and IL-2 for generating de novo Treg cells. The assessment of the expression of selected genes and proteins critical to Treg function and the proliferation assays were performed with the use of real-time RT-PCR and flow cytometry. RESULTS: After a 4-week stimulation with Treg expander and IL-2, the percentage of T-regulatory cells was significantly higher compared to the cells treated with medium alone (with no difference between diabetic and control children). However, we found some disturbances in the gene expression at mRNA level for molecules crucial for T-reg function. The induced Tregs from diabetic and control children were fully functional as assessed in proliferation assays. IN SUMMARY: Despite some disturbances at mRNA level in the critical gene expression, the suppressive properties of induced Treg cells from diabetic and control children were effective.


Assuntos
Diferenciação Celular , Diabetes Mellitus Tipo 1/imunologia , Linfócitos T Reguladores/fisiologia , Adolescente , Idade de Início , Algoritmos , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD4-Positivos/patologia , Linfócitos T CD4-Positivos/fisiologia , Diferenciação Celular/imunologia , Proliferação de Células , Células Cultivadas , Criança , Diabetes Mellitus Tipo 1/diagnóstico , Diabetes Mellitus Tipo 1/epidemiologia , Diabetes Mellitus Tipo 1/patologia , Feminino , Humanos , Subunidade alfa de Receptor de Interleucina-2/metabolismo , Ativação Linfocitária/fisiologia , Masculino , Cultura Primária de Células , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/metabolismo , Linfócitos T Reguladores/patologia
4.
Scand J Immunol ; 71(2): 115-22, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-20384863

RESUMO

The metabolic syndrome (MS) is defined as a cluster of risk factors, including abdominal obesity, dyslipidaemia, glucose intolerance and hypertension, which increase the risk for coronary heart disease. The immunological aspects of obesity and MS, including the role of T regulatory cells, have been intensively investigated. The aim of this study was to determine whether there is any disturbance in T regulatory cells number and/or function in children with MS. The percentages of T regulatory cells in the peripheral blood of children fulfilling the International Diabetes Federation criteria of the disease (n = 47) were assessed. Treg cells were also separated for further analysis of multiple genes important in their function with the use of real-time RT-PCR. We did not observe any difference in Treg percentages between study and control group but there was lower expression of some molecules including transforming growth factor-beta and interleukin-12 family members in Treg cells separated from children with MS compared to the healthy subjects. Our study is the first to report significant disturbances in some gene expression in T regulatory cells separated from children with MS. The results should be useful for further research in this field, including immunotherapeutic interventions.


Assuntos
Perfilação da Expressão Gênica , Síndrome Metabólica/genética , Síndrome Metabólica/imunologia , Linfócitos T Reguladores/imunologia , Separação Celular , Criança , Citometria de Fluxo , Expressão Gênica , Humanos , Síndrome Metabólica/sangue , Reação em Cadeia da Polimerase Via Transcriptase Reversa
5.
Neoplasma ; 56(5): 428-34, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-19580345

RESUMO

In all types of leukemia both in children and adults there is a need for novel therapies that could reduce the risk of relapse after standard treatment. Acute lymphoblastic leukemia (ALL) cells are ineffective antigen presenting cells, but as shown by many authors including results from our laboratory, stimulation with CD40L restores their antigen expressing capacity. The development of T-cell therapies for leukemic patients can be based on discovery of leukemia-associated antigens (LAA) which could be recognized by the host immune system. The aim of our present study was to test the hypothesis that leukemia-derived dendritic cells maintain the expression of tumor associated antigens. Twenty five children with B-cell precursor ALL were prospectively enrolled into the study. The mononuclear cells from peripheral blood or bone marrow were cultured and stimulated (or not) with CD40L and IL-4. The assessment of costimulatory/adhesion molecules with the use of flow cytometry and real-time RT PCR were used to confirm the possibility of turning ALL cells into dendritic-like cells. Additionally 22 tumor associated antigens mRNA levels were determined by real-time PCR technique with the TaqMan chemistry using ready-to-use Low Density Arrays for Gene Expression. The results of the study showed maintained expression and even up-regulation of some (PNPT1, PMPCB, HMMR/RHAMM, BSG and ERCC1) tumor associated antigens in CD40-activated leukemic cells. CD40L stimulation leading to the differentiation of leukemic cells into DCs which combine both antigen presenting function and expression of tumor associated antigens represents an interesting approach in cancer immunotherapy.


Assuntos
Antígenos de Neoplasias/genética , Basigina/genética , Proteínas de Ligação a DNA/genética , Células Dendríticas/metabolismo , Endonucleases/genética , Exorribonucleases/genética , Proteínas da Matriz Extracelular/genética , Receptores de Hialuronatos/genética , Metaloendopeptidases/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/imunologia , Ligante de CD40/farmacologia , Criança , Pré-Escolar , Feminino , Humanos , Imunoterapia , Interleucina-4/farmacologia , Masculino , Leucemia-Linfoma Linfoblástico de Células Precursoras/terapia , RNA Mensageiro/análise , Peptidase de Processamento Mitocondrial
6.
Eur J Med Res ; 14 Suppl 4: 165-9, 2009 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-20156750

RESUMO

OBJECTIVE: It has recently been described that circulatory and BAL regulatory T-cells (Tregs), defined as CD4+CD25highCD127low are increased in patients with active sarcoidosis compared with other interstitial lung diseases. MATERIAL AND METHODS: We studied prospectively 17 patients (10 women, 7 men) of median age 39 years (range 27-65) with active granulomatous lung diseases (GLD) (10 patients with sarcoidosis (BBS), and 7 with hypersensitivity pneumonitis (HP), and 9 healthy controls. Bronchoalveolar lavage fluid (BAL) and induced sputum Treg counts, CD4+, CD8+, CD25+ cells were quantified by flow cytometry. Disease activity was measured by ACE serum level. Pulmonary function tests were performed using an Elite DL Medgraphics body box. RESULTS: We found Treg cells count significantly elevated in induced sputum from active GLD (38.3% vs. 7.1% and 5.3% in BBS, HP, and control, respectively). A significantly higher percentage of Treg cells characterized BAL cells from HP patients (2.27%; 9.5%; 2.1%, in BBS, HP and control, respectively). There was a strong correlation with ACE serum level and Treg cell count in BAL fluid of BBS patients, with no such correlation within HP patient group, nor Treg cell count and pulmonary function tests. CONCLUSIONS: Our data suggest a potential role of CD4+CD25highCD127low induced sputum and BAL lymphocytes from patients with active granulomatous lung diseases and hypersensitivity pneumonitis. An increased number of Treg cells in active GLD may be involved in immune regulation in active granulomatous lung diseases. The results indicate that analysis of these cells could be useful as markers of disease activity in granulomatous lung diseases.


Assuntos
Líquido da Lavagem Broncoalveolar/imunologia , Sarcoidose Pulmonar/imunologia , Escarro/imunologia , Adulto , Idoso , Relação CD4-CD8 , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Linfócitos T Reguladores
7.
Autoimmunity ; 41(2): 163-73, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18324486

RESUMO

Apoptosis, i.e. natural programmed cell death, is a physiological phenomenon indispensable for normal functioning of the organism. The signal to apoptosis can be started practically in any cell. Disturbances in the apoptosis regulation determine the essential link of the pathogenesis of many diseases, including autoimmune thyroid disorders. The aim of the study was to assess the expression of Fas/FasL and caspase eight in the tissues of the thyroid gland in patients with Graves' disease (GD), non-toxic nodular goiter (NTNG) and Hashimoto's thyroiditis (HT). The analysis of Fas/FasL expression was performed by western blot and immunohistochemical investigation with DAB-visualization and Mayer's hematoxylin staining. Caspase-8 expression in thyroid follicular cells was assayed by western blot method. Identification of the proapoptotic proteins FasL and Fas exhibited their pronounced expression in the thyroid tissue in GD patients (++; ++) and HT (+++; +++) as compared to the NTNG group (0/+; 0/+). Among the study groups, the expression of caspase-8 was revealed in band 55 kDa from patients with autoimmune thyroid diseases. In GD patients, the percentage of thyrocytes with FasL expression correlated positively with TRAb (R = 0.58, p < 0.02). However, no such correlations were noted in HT or non-toxic multinodular goiter. There were no significant correlations between thyroid hormones and the percentage of thyrocytes with Fas and FasL expression. In conclusion, our findings suggest that the changes in the expression of apoptotic molecules on the surface of T lymphocytes and thyroid follicular cells in patients with autoimmune thyroid disorders reflect their substantial involvement in the pathogenesis of GD and HT. In addition, analysis of Fas/FasL and caspase-8 expression in thyroid tissue may indicate the disease activity and immunological phenotype.


Assuntos
Apoptose , Caspase 8/metabolismo , Proteína Ligante Fas/metabolismo , Doença de Graves/fisiopatologia , Doença de Hashimoto/fisiopatologia , Glândula Tireoide/metabolismo , Receptor fas/metabolismo , Adolescente , Adulto , Criança , Feminino , Bócio Nodular/metabolismo , Doença de Graves/metabolismo , Doença de Hashimoto/metabolismo , Humanos , Masculino , Glândula Tireoide/citologia
8.
J Physiol Pharmacol ; 59 Suppl 6: 499-505, 2008 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-19218674

RESUMO

The aim was to find out how the IL-12 and IL-18 levels in the bronchoalveolar lavage fluid (BALF) correspond to the inflammatory activity of the hypersensitivity pneumonitis (HP). We studied 12 patients with HP and 13 normal subjects. IL-12 and IL-18 levels were measured using ELISA kits. We found a significantly higher plasma angiotensin-converting enzyme (ACE) concentration (55 vs. 34 U/L, P=0.0016), lymphocyte percentage (57 vs. 14%, P<0.001), CD8+ cells (32 vs. 17%, P<0.001) and a lower CD4/CD8 ratio (1.2 vs. 2.0, P<0.0001). The IL-12 and IL-18 levels in BALF were significantly higher in HP patients than in healthy subjects (3.9 vs. 3.2 pg/ml, P=0.003 and 14.2 vs. 6.15 pg/ml, P% 0.0001, respectively). We found a strong positive correlation between IL-12 and the percentage of lymphocytes (r=0.68, P=0.015) and a negative one between IL-12 and the percentage of macrophages in BALF (r=-0.64, P=0.024). We conclude that upregulation of the Th1 cell cytokine profile may play a significant role in the pathogenesis of HP.


Assuntos
Alveolite Alérgica Extrínseca/metabolismo , Alveolite Alérgica Extrínseca/patologia , Líquido da Lavagem Broncoalveolar/citologia , Citocinas/biossíntese , Células Th1/metabolismo , Adulto , Broncoscopia , Eletrocardiografia , Feminino , Citometria de Fluxo , Humanos , Interleucina-12/metabolismo , Interleucina-18/metabolismo , Masculino , Pessoa de Meia-Idade , Oxigênio/sangue , Testes de Função Respiratória
9.
J Physiol Pharmacol ; 59 Suppl 6: 507-13, 2008 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-19218675

RESUMO

We studied prospectively 43 patients with sarcoidosis and 13 normal subjects. IL-12 and IL-18 levels were measured using ELISA kits. Spirometry and body plethysmography were performed using an Elite DL Medgraphics body box. The sarcoidosis group was characterized by significantly higher median range of the plasma angiotensin-converting enzyme (ACE) concentration (72 vs. 34 U/l, P<0.0001), lymphocyte (34 vs. 14%, P<0.0001), CD4+ cells percentages (59 vs. 36%), and CD4/CD8 ratio (4.2 vs. 1.99, P<0.0001). The BALF IL-12 and IL-18 levels were significantly higher in sarcoidosis patients than in healthy subjects (4.1 pg/ml vs. 3.2 pg/ml; P<0.001 and 11.1 pg/ml vs. 6.15 pg/ml, P<0.0001, respectively). A negative correlation between BALF IL-12 and ACE plasma levels (r=-0.33, P<0.05) within the sarcoidosis group was found. Our data suggest a potential role of IL-12 and IL-18 in a local immunologic response in pulmonary sarcoidosis.


Assuntos
Líquido da Lavagem Broncoalveolar/química , Interleucina-12/metabolismo , Interleucina-18/metabolismo , Sarcoidose Pulmonar/metabolismo , Adulto , Eletrocardiografia , Ensaio de Imunoadsorção Enzimática , Feminino , Citometria de Fluxo , Humanos , Masculino , Pessoa de Meia-Idade , Oxigênio/sangue , Peptidil Dipeptidase A/sangue , Pletismografia Total , Testes de Função Respiratória , Sarcoidose Pulmonar/patologia , Espirometria
10.
Neoplasma ; 54(6): 527-35, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17949237

RESUMO

UNLABELLED: Despite discovery of new therapeutic agents, including nucleoside analogs and monoclonal antibodies, the B-cell chronic lymphocytic leukemia (B-CLL) remains incurable. In recent years, some effort has been made in developing T-cell specific immunity against neoplasmatic cells. Reconstitution of effective costimulation and immunological response of host T-cells against CLL cells could be a potential approach in immunotherapeutic trials. CD40/CD40L system is involved in the survival and proliferation of normal and neoplasmatic B-cells. Some preclinical studies have shown that CD40 stimulation can differentiate leukemic cells into dendritic cells (DCs) and result in host response. In this study, we sought to determine whether B-CLL cells could be turned into efficient and functional antigen presenting cells, as well as to assess the type of allogeneic T-cell response against B-CLL - derived DCs. MATERIAL AND METHODS: B-CLL cells from 25 patients were cultured with or without the presence of CD40L and IL-4 for 96 hours and then cultured in mixed lymphocyte reaction with allogeneic T-cells. RESULTS: 1) after CD40 stimulation B-CLL cells achieved phenotypical and functional characterization of DCs (i.e. upregulated co-stimulatory and adhesion molecules at mRNA and protein level) 2) leukemia-derived DCs expressed higher amount of mRNA for chemokines involved in T-cell migration (MDC, TARC and CCR7) 3) the proliferating response of Tcells against leukemia-derived DCs consisted of CD4 and CD8 cells (upregulation of HLA-DR and OX40). CONCLUSIONS: our experiment confirm that B-CLL cells can be turned into dendritic-like cells, additionally, these cells express chemokines involved in T-cell migration and stimulate allogeneic response.


Assuntos
Quimiocinas/biossíntese , Quimiotaxia de Leucócito , Células Dendríticas/imunologia , Leucemia Linfocítica Crônica de Células B/imunologia , Ativação Linfocitária , Linfócitos T/imunologia , Idoso , Células Apresentadoras de Antígenos/citologia , Células Apresentadoras de Antígenos/imunologia , Ligante de CD40/metabolismo , Diferenciação Celular/imunologia , Células Cultivadas , Células Dendríticas/citologia , Feminino , Citometria de Fluxo , Antígenos HLA-DR/metabolismo , Humanos , Leucemia Linfocítica Crônica de Células B/terapia , Teste de Cultura Mista de Linfócitos , Masculino , Pessoa de Meia-Idade , Receptores OX40/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Linfócitos T/metabolismo , Transplante Homólogo
11.
J Physiol Pharmacol ; 58 Suppl 5(Pt 2): 445-52, 2007 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-18204157

RESUMO

In this study we compared the cellular profiles of induced sputum (IS) and BAL in newly diagnosed sarcoidosis patients before and after 6 months of prednisone therapy. The study encompassed 17 untreated patients with stage II pulmonary sarcoidosis. Sputum was induced with hypertonic saline solution, and bronchoscopy was performed in all patients. The results showed a significantly higher percentage of macrophages in BAL (P<0.05), whereas the percentage of neutrophils was higher in IS samples (P<0.01). The percentage of lymphocytes in IS was significantly lower than that in BAL (P<0.05). The TCD4/CD8 ratio was in favor of CD4 in both BAL and IS samples before prednisone therapy and it significantly decreased after therapy; the ratio was 4.8 vs. 1.8 (P=0.009) in BAL and 3.5 vs. 2.0 (P=0.019) in IS, respectively. Positive BAL/IS correlation characterized T cell populations in regards to CD4+ (r=0.59), CD8+ (r=0.34), CD4/CD8 ratio (r= 0.66) and CD3+ HLA-DR+ (r=0.89) cell population. These finding suggest that IS may serve as a valuable alternative to BAL technique.


Assuntos
Líquido da Lavagem Broncoalveolar/citologia , Sarcoidose Pulmonar/patologia , Escarro/citologia , Adulto , Idoso , Anti-Inflamatórios/uso terapêutico , Broncoscopia , Contagem de Células , Eletrocardiografia , Feminino , Citometria de Fluxo , Humanos , Masculino , Pessoa de Meia-Idade , Oxigênio/sangue , Prednisona/uso terapêutico , Testes de Função Respiratória , Sarcoidose Pulmonar/tratamento farmacológico , Espirometria
12.
Neoplasma ; 53(4): 301-4, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-16830056

RESUMO

UNLABELLED: Mechanisms leading blasts of acute lymphoblastic leukemia to escape from immune surveillance are still unknown. Only few reports showed that ALL cells are inefficient antigen presenting cells. The aim of the study was to assess expression of critical costimulatory/adhesion molecules and mRNA for main pro- and anti-inflammatory cytokines in ALL cells. Children with B-cell precursor ALL (n=20) were prospectively enrolled into the study. Expression of costimulatory/adhesion molecules (CD1a, CD11c, CD40, CD54, CD80, CD83, CD86, CD123, HLA class I and II) was assessed by flow cytometry and mRNA for cytokines (IFN-gamma, IL-10, IL-4, TGF-beta) - with real-time PCR. RESULTS: 1) high expression was observed for HLA I and II class, moderate for CD40, CD83, CD86 and low or no expression for CD80, CD54, CD1a, CD11c and CD123; 2) we found expression of mRNA for IFN-gamma, IL-10, IL-4 and TGF-beta in blasts cells (but not in all specimens). We noted relatively lower expression of all assessed cytokines comparing to T-cells obtained from healthy donors but interestingly expression for IL-10 was higher in normal B-cells than in blast cells, and IFN-gamma and IL-4 were not found in normal B-cells. In summary we suggest that ALL-blasts present low expression of costimulatory/adhesion molecules and mRNA for cytokines and this probably contribute to the absence of host T- cells stimulation to immune response.


Assuntos
Antígenos CD/metabolismo , Citocinas/metabolismo , Tolerância Imunológica , Leucemia-Linfoma Linfoblástico de Células Precursoras/imunologia , Leucemia-Linfoma Linfoblástico de Células Precursoras/metabolismo , Linfócitos B/metabolismo , Criança , Pré-Escolar , Citocinas/genética , Expressão Gênica , Humanos , Estudos Prospectivos , RNA Mensageiro/metabolismo
13.
Autoimmunity ; 38(8): 605-11, 2005 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-16390813

RESUMO

Thyroid peroxidase (TPO) is the major thyroid autoantigen recognized by serum autoantibodies from patients with Graves' disease (GD) or Hashimoto's thyroiditis directed to two immunodominant conformational regions termed A and B. The epitopes of human TPO have been defined using a panel of mouse monoclonal antibodies (mAbs). The aim of this study was to estimate the expression of chosen surface antigen regions of TPO (1, 18, 30, 64 epitopes) on thyroid cells in 15 patients with non-toxic multinodular goiter (NTMG) and 15 patients with GD. The thyrocytes were identified by indirect method: in the first stage we added mouse monoclonal autoantibodies specific for TPO regions and in the second stage we conjugated this complex with rabbit anti-mouse antibodies IgG (Fab')(2) with FITC. All investigations were performed by flow cytometry using Coulter EPICS XL apparatus. The percentages of thyrocytes with expression of epitopes 1, 18, 30, 64 TPO were measured in relation to the respective anti-TPO concentrations: 50-1600 microg/ml. The analysis of epitopes located in immunodominant regions (IDR) of TPO revealed higher percentages of thyrocytes in cases with GD in comparison to NTNG. The most predominant difference was observed for mAb 64 epitope (48 vs 7%, p < 0.019; 39 vs 5%, p < 0.017) at the concentration of 100-200 microg/ml mAbs. The expression of 18 epitope on thyrocytes was also statistically higher in Graves' patients than in the NTMG (14 vs 6%, p < 0.025) at concentration of 400 microg/ml mAbs. However, this expression was much less pronounced. In all the cases, the percentages of thyrocytes with epitopes 1 and 30 were in low detection (8-15% of positive cells). In conclusion, our findings suggest that the elevated expression of TPO epitopes 18 and 64 in young patients with thyroid autoimmune diseases increase stimulation and activation of thyroid cells during inflammatory reaction within the thyroid gland. In addition, predominant expression of 64 TPO epitope that recognizes B domain in GD patients could be a useful marker of the immune process in the thyroid gland.


Assuntos
Anticorpos Monoclonais , Bócio Nodular/imunologia , Doença de Graves/imunologia , Epitopos Imunodominantes/metabolismo , Iodeto Peroxidase/imunologia , Linfócitos T/imunologia , Adolescente , Animais , Biomarcadores/análise , Feminino , Citometria de Fluxo , Bócio Nodular/enzimologia , Doença de Graves/enzimologia , Humanos , Epitopos Imunodominantes/imunologia , Iodeto Peroxidase/metabolismo , Masculino , Camundongos , Linfócitos T/enzimologia
15.
Rocz Akad Med Bialymst ; 49 Suppl 1: 37-9, 2004.
Artigo em Inglês | MEDLINE | ID: mdl-15638368

RESUMO

The aim of this study was to check if the expression of CD44v4 in epithelial cells of the colorectal cancer correlates with the pTN stage and the histopathological grade of malignancy--G. Samples of tumour tissue (TT), as well as those of healthy tissue (HT) and of tumour adjacent tissue (TAT) were obtained from 25 patients. An evaluation of the expression of CD44v4 was performed in a flow cytometer. The mean value of the percentage of epithelial cells with co-expression of CD44v4 was lower in pT2 stage than that in pT3 only in HT. The expression of CD44v4 in epithelial cells was higher in cases without lymph node metastases only in TAT. The expression of CD44v4 in epithelial cells was higher in G2 than in G3 degree only in TAT as well. According to the obtained results, it is difficult to state if CD44v4 can influence the progress of colorectal cancer.


Assuntos
Neoplasias Colorretais/imunologia , Variação Genética , Receptores de Hialuronatos/genética , Mucosa Intestinal/imunologia , Antígenos CD/genética , Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , Citometria de Fluxo , Humanos , Mucosa Intestinal/patologia
16.
Rocz Akad Med Bialymst ; 49 Suppl 1: 34-6, 2004.
Artigo em Inglês | MEDLINE | ID: mdl-15638367

RESUMO

The aim of the study was to assess the expression of adhesion molecules, CD134 and CD137, in the peripheral blood in correlation with clinical advancement, the histological grade, the size and type of tumour growth, tissue infiltration, and lymph node and metastases to lymph nodes and liver. The study involved 28 patients with primary colorectal cancer. The expression of both molecules was investigated on the day of the surgery, before the procedure and ten days after the operation by means of flow cytometry. The expression of CD134 was markedly higher, compared to CD137, both on the day of the surgery and ten days after the operation. A significant increase was observed in CD134 expression ten days after the surgery. CD137 expression increased with the higher stage of clinical advancement, but decreased with the enhancement of colon wall infiltration. CD134 showed a similar expression for all the stages of tumour clinical advancement.


Assuntos
Neoplasias Colorretais/patologia , Receptores de Fator de Crescimento Neural/sangue , Receptores do Fator de Necrose Tumoral/sangue , Antígenos CD , Neoplasias Colorretais/sangue , Neoplasias Colorretais/imunologia , Neoplasias Colorretais/cirurgia , Citometria de Fluxo , Humanos , Estadiamento de Neoplasias , Período Pós-Operatório , Receptores OX40 , Membro 9 da Superfamília de Receptores de Fatores de Necrose Tumoral
17.
Rocz Akad Med Bialymst ; 49 Suppl 1: 91-3, 2004.
Artigo em Inglês | MEDLINE | ID: mdl-15638386

RESUMO

The aim of this study was to evaluate the expression of CD44v10 in colorectal tumour cells and in lymphocytes infiltrating the tumour (CD45+). Samples of tumour tissue (TT), as well as of healthy tissue (HT) and of tumour adjacent tissue (TAT), were obtained from 20 patients. An evaluation of CD44v10 expression was performed in a flow cytometer. The mean value of the percentage of CD45+ with co-expression of CD44v10 was significantly higher in the lower stage of the tumour (pT). The mean value of the percentage of epithelial cells with CD44v10 co-expression was significantly higher in pN2 than in pN1 stage. Only in TAT the mean value of the percentage of epithelial cells and CD45+ with tCD44v10 co-expression was significantly lower in the higher degree of histological malignancy. It is supposed that CD44v10 takes part in local cancer progression.


Assuntos
Neoplasias Colorretais/genética , Variação Genética , Receptores de Hialuronatos/genética , Linfócitos do Interstício Tumoral/patologia , Neoplasias Colorretais/patologia , Humanos , Linfócitos do Interstício Tumoral/imunologia , Recidiva Local de Neoplasia
18.
Rocz Akad Med Bialymst ; 49: 106-10, 2004.
Artigo em Inglês | MEDLINE | ID: mdl-15631324

RESUMO

PURPOSE: The aim of the study was to assess the expression of selected adhesion molecules on mononuclear cells of peripheral blood and lymphocyte subpopulations in children with IgA nephropathy (IgAN). MATERIAL AND METHODS: 14 children with IgAN and 20 healthy controls were included in the study. Flow cytometry was used to determine the expression of such adhesion molecules as L selectin (CD62L), VLA-4 integrin (CD49d), intracellular molecule ICAM-1 (CD54) and cytotoxic lymphocyte molecule CTLA-4 (CD152), as well as the lymphocyte antigens: CD3, CD4, CD8, CD19, CD1656 (NK), CD4 and CD8 RO+ and RA+. RESULTS: The findings revealed that the expression of the adhesion molecules VLA-4 and CTLA-4 did not differ from that of the healthy controls (p > 0.05). However, the expression of CD62L (L-selectin) was increased (p < 0.05). The expression of ICAM-1 was reduced, but not significantly, compared to the control group (p > 0.05). We found a decrease in the expression of NK cells (CD1656) and CD4/CD8 ratio, and an increase in CD8 cells (p < 0.05). In the group of 9/14 children, with proteinuria over 1.0 g/24 hours, a decreased expression of CD4 was additionally found (p < 0.05). CONCLUSIONS: The children with IgAN show: 1. Changes in peripheral lymphocyte subpopulations involving an increase in CD8 cells and a decrease in CD1656(NK) cells, a reduction in the CD4/CD8 ratio, and additionally in cases with proteinuria a reduction in CD4 cell count, 2. Increased expression of L-selectin (CD62L) on peripheral blood mononuclear cells.


Assuntos
Moléculas de Adesão Celular/sangue , Glomerulonefrite por IGA/sangue , Subpopulações de Linfócitos , Adolescente , Antígenos CD , Antígenos CD19/sangue , Antígenos de Diferenciação/sangue , Complexo CD3/sangue , Antígenos CD4/sangue , Relação CD4-CD8 , Antígeno CD56/sangue , Antígenos CD8/sangue , Antígeno CTLA-4 , Estudos de Casos e Controles , Criança , Feminino , Expressão Gênica , Humanos , Integrina alfa4beta1/sangue , Molécula 1 de Adesão Intercelular/sangue , Selectina L/sangue , Masculino , Receptores de IgG/sangue
19.
Rocz Akad Med Bialymst ; 48: 95-9, 2003.
Artigo em Inglês | MEDLINE | ID: mdl-14737951

RESUMO

PURPOSE: Helicobacter pylori infection in children is associated with a chronic inflammatory process of gastric and duodenal mucosa, which may have a various clinical course ranging from asymptomatic and chronic inflammatory condition to gastric ulceration. The immune system may contribute especially to chronic gastric mucosa inflammation. The aim of our study was to assess the levels of peripheral blood T (CD3+, CD4+, CD8+) and B lymphocyte subpopulation (CD19+) in children with Helicobacter pylori infection and to evaluate their relation to degree of antrum mucosa inflammation. MATERIAL AND METHODS: The study was performed in 32 children aged 7-18 years, hospitalized due to dyspeptic symptoms. The endoscopic examination of upper gastrointestinal tract was performed and gastric and duodenal mucosa was estimated in all patients. The endoscopic and histological evaluation of gastric mucosa was performed according to the Sydney System [4]. The urease test (CLO-test-H. pylori) was made to estimate the severity of the infection. RESULTS: Moderate antrum mucosa inflammation was found in 41.2% of the examined. The highest percentage of children (58.8%) presented marked inflammation. No mild inflammation was found in children examined. CONCLUSIONS: No correlation was found between lymphocyte levels and the degree of the inflammatory changes in antrum mucosa. The evaluation of peripheral blood lymphocytes performed in children with Helicobacter pylori infection suggests that T lymphocytes may play a predominant role in this infection.


Assuntos
Infecções por Helicobacter/imunologia , Helicobacter pylori , Linfócitos/sangue , Adolescente , Criança , Dispepsia/imunologia , Feminino , Mucosa Gástrica/imunologia , Infecções por Helicobacter/sangue , Humanos , Linfócitos/imunologia , Masculino , Índice de Gravidade de Doença
20.
Pol Merkur Lekarski ; 10(58): 244-6, 2001 Apr.
Artigo em Polonês | MEDLINE | ID: mdl-11434167

RESUMO

The main lymphocytes' subpopulations of peripheral blood in 21 children (mean age--9.8 +/- 3.3 y) with permanent proteinuria and haematuria, 11 with IgA nephropathy (IgA) and 10 with nephropathy in course of Schonlein-Henoch purpura (Sch-H) during intensification of symptoms were assessed on flow cytometer f. Coultier, using monoclonal antibodies. The control group consisted of 21 healthy children at the same age. The results showed decreased percentage of CD4 and increased percentage of CD8 with decreased CD4/CD8 ratio in both groups of examined children. In most cases also increased percentage of B lymphocytes (CD19) were noticed and especially in children from N IgA group increased concentration of immunoglobulin A in serum. Only in some patients increased percentage of natural cytotoxic cells NK were noticed. The results were similar either in N IgA or Sch-H groups.


Assuntos
Glomerulonefrite por IGA/imunologia , Vasculite por IgA/imunologia , Subpopulações de Linfócitos/imunologia , Adolescente , Linfócitos B/imunologia , Relação CD4-CD8 , Criança , Humanos , Imunoglobulina A/sangue , Contagem de Linfócitos
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