RESUMO
In HSCT setting, KIR-driven alloreactivity might be better predicted if the donor KIR genotype is considered in addition to the recipient HLA genotype. The prediction of NK cell alloreactivity relies on the missing ligand in the recipient, a scenario that can be found in HLA-identical and non-identical allotransplants. The aim of this study was to investigate at genetic level the prognostic impact of recipient HLA-I lacking for donor KIR on allotransplanted patients outcome. We analysed donors KIR genotype and HLA genotype of 60 paediatric patients who received related (n=15) or unrelated (n=45) transplantation. When patients were grouped based on the KIR gene type involved in the KIR/HLA-I mismatch, we did not observe any relapse in the group of patients characterized by mismatches involving only inhibitory KIR. On the contrary, all relapses were observed in patients showing at least one activating gene involved in the mismatch (p<0.05). Although the biological mechanism accounting for this putative genetic rule is still to be clarified, we suggest that a careful survey of KIR/HLA-I mismatching should be taken into account in the selection of donor in related and unrelated HSCT.
Assuntos
Antígeno HLA-A1/genética , Neoplasias Hematológicas/cirurgia , Transplante de Células-Tronco Hematopoéticas/métodos , Receptores KIR/genética , Adolescente , Criança , Pré-Escolar , Estudos de Coortes , Reparo de Erro de Pareamento de DNA/genética , Intervalo Livre de Doença , Feminino , Seguimentos , Genótipo , Rejeição de Enxerto/genética , Antígeno HLA-A1/análise , Haploidia , Neoplasias Hematológicas/mortalidade , Neoplasias Hematológicas/patologia , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Histocompatibilidade/genética , Humanos , Masculino , Modelos de Riscos Proporcionais , Receptores KIR/análise , Recidiva , Medição de Risco , Estatísticas não Paramétricas , Taxa de Sobrevida , Transplante Homólogo , Resultado do TratamentoRESUMO
OBJECTIVE: Endothelial progenitor cells (EPCs) participate in vascular homeostasis and angiogenesis. The aim of the present study was to explore EPC number and function in relation to cardiovascular risk, gender, and reproductive state. METHODS AND RESULTS: As measured by flow-cytometry in 210 healthy subjects, CD34(+)KDR(+) EPCs were higher in fertile women than in men, but were not different between postmenopausal women and age-matched men. These gender gradients mirrored differences in cardiovascular profile, carotid intima-media thickness, and brachial artery flow-mediated dilation. Moreover, EPCs and soluble c-kit ligand varied in phase with menstrual cycle in ovulatory women, suggesting cyclic bone marrow mobilization. Experimentally, hysterectomy in rats was followed by an increase in circulating EPCs. EPCs cultured from female healthy donors were more clonogenic and adherent than male EPCs. Treatment with 17beta-estradiol stimulated EPC proliferation and adhesion, via estrogen receptors. Finally, we show that the proangiogenic potential of female EPCs was higher than that of male EPCs in vivo. CONCLUSIONS: EPCs are mobilized cyclically in fertile women, likely to provide a pool of cells for endometrial homeostasis. The resulting higher EPC levels in women than in men reflect the cardiovascular profile and could represent one mechanism of protection in the fertile female population.
Assuntos
Doenças Cardiovasculares/etiologia , Endotélio Vascular/metabolismo , Endotélio Vascular/patologia , Estrogênios/metabolismo , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/patologia , Caracteres Sexuais , Adulto , Idoso , Animais , Ataxina-1 , Ataxinas , Doenças Cardiovasculares/metabolismo , Doenças Cardiovasculares/patologia , Adesão Celular/fisiologia , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Estradiol/farmacologia , Feminino , Homeostase/fisiologia , Humanos , Recém-Nascido , Masculino , Ciclo Menstrual/metabolismo , Pessoa de Meia-Idade , Neovascularização Fisiológica/fisiologia , Proteínas do Tecido Nervoso/metabolismo , Proteínas Nucleares/metabolismo , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Proteínas Proto-Oncogênicas c-kit/metabolismo , Ratos , Receptores de Estrogênio/metabolismo , Fatores de RiscoRESUMO
OBJECTIVE: Our purpose was to study in vitro whether phenotypically-distinct interstitial cell clones from bovine aortic valve (BVIC) possess different calcifying potential in response to endotoxin (lipopolysaccharide [LPS]) and phosphate (Pi). METHODS AND RESULTS: Among various clones of BVIC obtained by limited dilution technique we selected 4 clones displaying different growth patterns and immunophenotypes. Uncloned and cloned cells were treated with combinations of LPS (100 ng/mL) and Pi (2.4 mmol/L). Uncloned BVIC showed increased alkaline phosphatase activity (ALP) after treatment with LPS, which resulted in calcification after addition of Pi. Among BVIC clones, only Clone 1 (fibroblast-like phenotype) showed a relevant increase in ALP after LPS treatment in parallel with prevention of smooth muscle (SM) alpha-actin accumulation. No effect was observed in clonal cells harboring a more stable SM cell-like profile (Clone 4). None of the isolated clones calcified but mineralization was induced in the presence of LPS plus Pi when Clone 1 was cocultured with Clone 4 or after seeding on type I collagen sponges. CONCLUSIONS: Endotoxin and phosphate can act as valve calcification promoters by targeting specific fibroblast-like interstitial valve cells that possess a unique procalcific potential.
Assuntos
Valva Aórtica/efeitos dos fármacos , Valva Aórtica/patologia , Calcinose/etiologia , Doenças das Valvas Cardíacas/etiologia , Lipopolissacarídeos/toxicidade , Fosfatos/toxicidade , Fosfatase Alcalina/metabolismo , Animais , Valva Aórtica/metabolismo , Calcinose/metabolismo , Calcinose/patologia , Cálcio/metabolismo , Bovinos , Células Clonais , Técnicas de Cocultura , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Fibroblastos/patologia , Doenças das Valvas Cardíacas/metabolismo , Doenças das Valvas Cardíacas/patologia , Miócitos de Músculo Liso/efeitos dos fármacos , Miócitos de Músculo Liso/metabolismo , Miócitos de Músculo Liso/patologia , FenótipoRESUMO
OBJECTIVE: The loss of myenteric neurons in the lower esophageal sphincter (LES) characterizes achalasia, an esophageal motor disorder. Because the presence of lymphocytic infiltrates suggests an immuno-mediated mechanism ongoing at the sites of disease, we investigated the T-cell receptor (TCR) repertoire and the ability to recognize human herpes virus type 1 (HSV-1) antigens of LES-infiltrating T lymphocytes in achalasia patients. METHODS: Fifty-nine patients with idiopathic achalasia and 38 heart-beating cadaveric multiorgan donors (controls) were studied. By flow cytometry evaluation and CDR3 length spectratyping analysis, the lymphocytes of 18 patients and 15 controls were analyzed, whereas 41 patients and 23 controls were employed for functional assays. RESULTS: Achalasia patients were characterized by a significantly higher esophagus lymphocytic infiltrate than controls (24.71%+/- 3.11 and 9.54%+/- 1.34, respectively; P < 0.05), mainly represented by CD3+CD8+ T cells. The characterization of TCR beta chain repertoire of CD3+ cells showed the expression of a limited number of TCR beta variable (BV) gene families (from two to five out of 26), with highly restricted spectratypes, suggesting a disease-associated oligoclonal selection of T cells. Furthermore, lymphocytes from achalasia LES specifically responded to exposure to HSV-1 antigens in vitro as showed by increased proliferation and Th-1 type cytokines release. CONCLUSIONS: These data suggest that the oligoclonal lymphocytic infiltrate within the LES of achalasia patients may represent the trace of an immune-inflammatory reaction triggered by HSV-1 antigens and that the Th1-type cytokines released by the activated lymphocytes may contribute to establish the neuronal damage accounting for the clinical features of idiopathic achalasia.
Assuntos
Antígenos Virais/imunologia , Acalasia Esofágica/imunologia , Herpesvirus Humano 1/imunologia , Plexo Mientérico/imunologia , Receptores de Antígenos de Linfócitos T/análise , Linfócitos T/imunologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Cadáver , Esfíncter Esofágico Inferior/inervação , Feminino , Citometria de Fluxo , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da PolimeraseRESUMO
We have shown previously that the chemokine receptors CXCR3 and CXCR6 are coexpressed by Th1 cells infiltrating the lung and the granuloma of patients with sarcoidosis. In this study, we evaluated the role of CCL20/CCR6 interaction in the pathogenesis of acute and chronic pulmonary sarcoidosis. By flow cytometry and molecular analyses, we have demonstrated that Th1 cells isolated from the bronchoalveolar lavage (BAL) of patients with sarcoidosis and T cell alveolitis are equipped with CCR6. Furthermore, CCR6(+) T cells coexpressed the chemokine receptors CXCR3 and CXCR6. Immunohistochemical analysis of lung specimens has shown that CCR6(+) T cells infiltrate lung interstitium and surround the central core of the granuloma. It is interesting that CCR6 was never detected on the alveolar macrophage (AM) surface, and it is observed in the cytoplasm of AMs from patients with sarcoidosis and alveolitis. The CCR6 ligand CCL20 was expressed by macrophages, multinucleated giant cells, and epithelioid cells infiltrating the granuloma. Furthermore, detectable levels of CCL20 protein are seen in the BAL fluid components of patients with active sarcoidosis, and sarcoid AMs release the CCR6 ligand in vitro. From a functional point of view, sarcoid Th1 cells were able to respond to CXCL10, CXCL16, and CCL20 in migratory assays. In vitro kinetic studies demonstrated that CCR6 is induced rapidly by IL-2, IL-18, and IFN-gamma. In conclusion, T cells expressing CCR6, CXCR3, and CXCR6 act coordinately with respective ligands and Th1 inflammatory cytokines in the alveolitic/granuloma phases of the disease.
Assuntos
Quimiocina CCL20/imunologia , Regulação da Expressão Gênica/imunologia , Pulmão/imunologia , Proteínas Inflamatórias de Macrófagos/imunologia , Receptores CCR6/imunologia , Sarcoidose Pulmonar/imunologia , Doença Aguda , Adulto , Lavagem Broncoalveolar , Células Cultivadas , Quimiocina CCL20/biossíntese , Quimiocina CXCL10/biossíntese , Quimiocina CXCL10/imunologia , Quimiocina CXCL16 , Quimiocinas CXC/biossíntese , Quimiocinas CXC/imunologia , Doença Crônica , Células Epitelioides/imunologia , Células Epitelioides/metabolismo , Células Epitelioides/patologia , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Células Gigantes/imunologia , Células Gigantes/metabolismo , Células Gigantes/patologia , Granuloma do Sistema Respiratório/imunologia , Granuloma do Sistema Respiratório/metabolismo , Granuloma do Sistema Respiratório/patologia , Humanos , Interferon gama/imunologia , Interferon gama/farmacologia , Interleucina-18/imunologia , Interleucina-18/farmacologia , Interleucina-2/imunologia , Interleucina-2/farmacologia , Pulmão/metabolismo , Pulmão/patologia , Proteínas Inflamatórias de Macrófagos/biossíntese , Macrófagos Alveolares/imunologia , Macrófagos Alveolares/metabolismo , Macrófagos Alveolares/patologia , Masculino , Pessoa de Meia-Idade , Receptores CCR6/biossíntese , Receptores CXCR3 , Receptores CXCR6 , Receptores de Quimiocinas/biossíntese , Receptores de Quimiocinas/imunologia , Receptores Depuradores/biossíntese , Receptores Depuradores/imunologia , Receptores Virais/biossíntese , Receptores Virais/imunologia , Sarcoidose Pulmonar/metabolismo , Sarcoidose Pulmonar/patologia , Células Th1/imunologia , Células Th1/metabolismoRESUMO
By analyzing the expression of several cytotoxic markers, killer-immunoglobulin-like receptors (KIRs), CD94/CD159, CD314 and natural cytotoxicity receptors (NCRs), in 22 CD3+ lymphoproliferative disease of granular lymphocyte (LDGL) patients we investigated whether granular lymphocytes (GLs) displayed the phenotype of fully differentiated cytotoxic cells. Our results demonstrate that GLs express a pattern consistent with fully differentiated CTLs. KIRs are expressed only in a fraction of patients (7/22), as is CD94/CD159 (5/22). In conclusion, GLs in CD3+ LDGL patients typically show the phenotype of fully differentiated CTL, whereas the expression of NK receptors does not represent a common feature of the proliferating clone.
Assuntos
Linfócitos/imunologia , Transtornos Linfoproliferativos/imunologia , Linfócitos T Citotóxicos/imunologia , Idoso , Complexo CD3/imunologia , Diferenciação Celular/imunologia , Feminino , Humanos , Imunofenotipagem , Masculino , Subfamília K de Receptores Semelhantes a Lectina de Células NK , Receptores de IgG/imunologia , Receptores Imunológicos/biossíntese , Receptores Imunológicos/imunologia , Receptores KIR , Receptores de Células Matadoras NaturaisRESUMO
OBJECTIVE: Peripheral arterial disease (PAD) is a threatening complication of diabetes. As endothelial progenitor cells (EPCs) are involved in neovasculogenesis and maintenance of vascular homeostasis, their impairment may have a role in the pathogenesis of diabetic vasculopathy. This study aimed to establish whether number and function of EPCs correlate with PAD severity in type 2 diabetic patients. METHODS AND RESULTS: EPCs were defined by the expression of CD34, CD133 and KDR, and quantified by flow cytometry in 127 diabetic patients with and without PAD. PAD severity has been assessed as carotid atherosclerosis and clinical stage of leg atherosclerosis obliterans. Diabetic patients with PAD displayed a significant 53% reduction in circulating EPCs versus non-PAD patients, and EPC levels were negatively correlated with the degree of carotid stenosis and the stage of leg claudication. Moreover, the clonogenic and adhesion capacity of cultured EPCs were significantly lower in diabetic patients with PAD versus patients without. CONCLUSIONS: This study demonstrates that EPC decrease is related to PAD severity and that EPC function is altered in diabetic subjects with PAD, strengthening the pathogenetic role of EPC dysregulation in diabetic vasculopathy. EPC count may be considered a novel biological marker of peripheral atherosclerosis in diabetes.
Assuntos
Angiopatias Diabéticas/patologia , Angiopatias Diabéticas/fisiopatologia , Células Endoteliais/patologia , Doenças Vasculares Periféricas/patologia , Doenças Vasculares Periféricas/fisiopatologia , Células-Tronco/patologia , Idoso , Artérias , Células Sanguíneas/patologia , Estenose das Carótidas/patologia , Estenose das Carótidas/fisiopatologia , Estudos de Casos e Controles , Adesão Celular , Contagem de Células , Células Cultivadas , Ensaio de Unidades Formadoras de Colônias , Diabetes Mellitus Tipo 2 , Feminino , Citometria de Fluxo , Humanos , Claudicação Intermitente/patologia , Claudicação Intermitente/fisiopatologia , Perna (Membro)/irrigação sanguínea , Masculino , Índice de Gravidade de DoençaRESUMO
BACKGROUND AND PURPOSE: Disruption of the endothelial layer is the first step in the atherogenic process. Experimental studies have shown that endothelial progenitor cells (EPCs) are involved in endothelial homeostasis and repair. Conversely, EPC depletion has been demonstrated in the setting of established atherosclerotic diseases. With this background, we evaluated whether variations in the number of EPCs are associated with subclinical atherosclerosis in healthy subjects. METHODS: Carotid intima-media thickness (IMT), high-sensitive C-reactive protein (hsCRP), levels of circulating EPCs, and cardiovascular risk were compared in 137 healthy subjects. Six subpopulations of progenitor cells were determined by flow cytometry on the basis of the surface expression of CD34, CD133, and KDR antigens: CD34(+), CD133(+), CD34(+)CD133(+), CD34(+)KDR(+), CD133(+)KDR(+), and CD34(+)CD133(+)KDR(+). RESULTS: Among different antigenic profiles of EPCs, only CD34(+)KDR(+) cells were significantly reduced in subjects with increased IMT. Specifically, CD34(+)KDR(+) cells were inversely correlated with IMT, even after adjustment for hsCRP and 10-year Framingham risk and independently of other cardiovascular parameters. CONCLUSIONS: Depletion of CD34(+)KDR(+) EPCs is an independent predictor of early subclinical atherosclerosis in healthy subjects and may provide additional information beyond classic risk factors and inflammatory markers.
Assuntos
Antígenos CD34/metabolismo , Antígenos de Superfície/metabolismo , Células Sanguíneas/metabolismo , Células Endoteliais/metabolismo , Arteriosclerose Intracraniana/sangue , Arteriosclerose Intracraniana/diagnóstico por imagem , Células-Tronco/metabolismo , Adulto , Artérias Carótidas/diagnóstico por imagem , Contagem de Células , Células Endoteliais/patologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Células-Tronco/patologia , Túnica Íntima/diagnóstico por imagem , Túnica Média/diagnóstico por imagem , UltrassonografiaRESUMO
OBJECTIVES: We sought to establish whether a reduction in endothelial progenitor cells (EPCs) has a putative role in peripheral vascular disease (PVD) of type 2 diabetic patients. BACKGROUND: Peripheral vascular disease is a common and severe complication of diabetes mellitus. Impaired collateralization of diabetic vasculopathy has been extensively shown, but causes leading to its pathogenesis are not fully understood. Recently, EPCs have been found to contribute to vascular repair and angiogenesis. Diabetes has been associated with low levels of circulating EPCs, but no data are available in the literature on the relationship between EPCs and PVD in diabetes. METHODS: Flow cytometric analysis was used to quantify circulating progenitor cells (CPCs, CD34+) and EPCs (CD34+KDR+) in 51 patients and 17 control subjects. RESULTS: The CPCs and EPCs from diabetic patients were reduced by 33% and 40%, respectively, compared with healthy subjects (p < 0.001). An inverse correlation was found between the number of EPCs and the values of fasting glucose (r = -0.49, p = 0.006). Peripheral vascular disease was associated with a 47% reduction in EPCs (p < 0.0001) and EPC levels directly correlated with the ankle-brachial index (r = 0.70, p = 0.01). The subgroup of diabetic patients with PVD also had reduced CPCs by 32% (p = 0.037), whereas patients with ischemic foot lesions had the lowest levels of both EPCs and CPCs (p = 0.02). CONCLUSIONS: Our data demonstrate decreased EPC levels in diabetic patients and, for the first time, show that PVD is associated with an extensively low number of EPCs. Depletion of circulating EPCs in diabetic patients may be involved in the pathogenesis of peripheral vascular complications.
Assuntos
Diabetes Mellitus Tipo 2/fisiopatologia , Angiopatias Diabéticas/patologia , Endotélio Vascular/citologia , Doenças Vasculares Periféricas/patologia , Idoso , Antígenos CD34/análise , Estudos de Casos e Controles , Angiopatias Diabéticas/sangue , Feminino , Citometria de Fluxo , Hemoglobinas Glicadas/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Doenças Vasculares Periféricas/sangueRESUMO
A role for circulating progenitor cells have been recently demonstrated in many pathological conditions. Endothelial progenitor cells (EPCs) localize at sites of ischemia and stimulate neovasculogenesis. This small study was carried out to assess whether selective blood sampling during renal angiography could demonstrate an arterio-venous gradient of EPCs in 5 patients with renal artery stenosis. Surprisingly, we found that EPCs were more abundant in venous than in arterial blood, suggesting that the kidney subjected to chronic ischemia may become a source rather than a target of EPCs.
Assuntos
Endotélio Vascular/patologia , Hipertensão Renal/patologia , Obstrução da Artéria Renal/patologia , Artéria Renal , Veias Renais , Células-Tronco/patologia , Humanos , Isquemia/patologia , Rim/irrigação sanguínea , Rim/patologiaRESUMO
BACKGROUND: Hypersensitivity pneumonitis (HP) is an interstitial lung disease caused by repeated inhalations of finely dispersed organic particles or low molecular weight chemicals. The disease is characterized by an alveolitis sustained by CD8(+) cytotoxic T lymphocytes, granuloma formation, and, whenever antigenic exposition continues, fibrosis. Although it is known that T-cell migration into the lungs is crucial in HP reaction, mechanisms implicated in this process remain undefined. METHODS: Using flow cytometry, immunohistochemistry, confocal microscopy analysis and chemotaxis assays we evaluated whether CXCL10 and its receptor CXCR3 regulate the trafficking of CD8(+) T cells in HP lung. RESULTS: Our data demonstrated that lymphocytes infiltrating lung biopsies are CD8 T cells which strongly stain for CXCR3. However, T cells accumulating in the BAL of HP were CXCR3(+)/IFNgamma(+) Tc1 cells exhibiting a strong in vitro migratory capability in response to CXCL10. Alveolar macrophages expressed and secreted, in response to IFN-gamma, definite levels of CXCL10 capable of inducing chemotaxis of the CXCR3(+) T-cell line. Interestingly, striking levels of CXCR3 ligands could be demonstrated in the fluid component of the BAL in individuals with HP. CONCLUSION: These data indicate that IFN-gamma mediates the recruitment of lymphocytes into the lung via production of the chemokine CXCL10, resulting in Tc1-cell alveolitis and granuloma formation.
Assuntos
Alveolite Alérgica Extrínseca/imunologia , Linfócitos T CD8-Positivos/imunologia , Quimiocinas CXC/imunologia , Interferon gama/imunologia , Ativação Linfocitária/imunologia , Receptores de Quimiocinas/imunologia , Adulto , Alveolite Alérgica Extrínseca/patologia , Movimento Celular/imunologia , Quimiocina CXCL10 , Feminino , Humanos , Masculino , Receptores CXCR3RESUMO
PURPOSE: The chronic exposure at high altitude (HA) represents an ideal model for evaluating the in vivo effects of hypobaric hypoxia. Taking advantage of the EV-K2-CNR Pyramid, this study was designed to evaluate whether acute and chronic hypoxia differently modulates the in vivo immune responses. METHODS: The study includes 13 healthy female moderately active volunteers participating to the Italian HA project EV-K2-CNR. Peripheral blood lymphocytes, collected at sea level and at HA in the Pyramid Laboratory of CNR, Nepal (5050 m), were immunologically characterized by flow cytometry and a series of molecular and functional analyses. RESULTS: Flow cytometric analyses showed that: a) CD3+ T lymphocytes significantly decreased during both acute and chronic exposure to HA, b) T-cell fall was totally due to CD4+ T-cell reduction, c) B lymphocytes were not influenced by the exposure to HA, and d) natural killer (NK) cells significantly increased during acute and chronic exposure. The evaluation of the Th1/Th2 pattern demonstrated a significant decrease of the expression of the Th1 cytokine interferon-gamma (IFN-gamma) by circulating T cells during acute and chronic exposure to HA. The expression by T cells of CXCR3, a chemokine receptor typically expressed by Th1/Tc1 cells, paralleled the decrease of IFN-gamma. On the contrary, the expression of IL-4 was not conditioned by the exposure to HA. Finally, functional studies showed a significant reduction of the proliferative activity in response to mitogen (PHA) both in acute and chronic HA exposure. Despite the increased number of NK cells, NK cytotoxic activity was not influenced by the HA exposure. CONCLUSIONS: Our results indicate that the in vivo exposure to HA leads to an impairment of the homeostatic regulation of Th1/Th2 immune balance that potentially could favor long-term immunological alterations and increase the risk of infections.
Assuntos
Altitude , Hipóxia/metabolismo , Adulto , Complexo CD3/sangue , Antígenos CD4/sangue , Proliferação de Células , Células Cultivadas , Feminino , Citometria de Fluxo , Humanos , Interferon gama/sangue , Interleucina-4/sangue , Células Matadoras Naturais/metabolismo , Receptores CXCR3 , Receptores de Quimiocinas/sangue , Linfócitos T/metabolismo , Células Th1/metabolismoRESUMO
Hypersensitivity pneumonitis (HP) is characterized by an alveolitis sustained by CD8(+) T lymphocytes showing a limited expression of the T cell receptor (TCR). We previously demonstrated that a bias in T cell selection occurs in the lower respiratory tract of patients with HP, with a compartmentalization in the lung of CD8(+) T cells bearing (TCR)-beta variable (TCRBV) #2, 3, 5, 6, 8, and 13 gene segments. We herein characterized the clonal T cell populations present in the lung and in the blood of patients with HP. Heteroduplex analyses, cloning, and sequencing T cells bearing TCR indicate oligoclonal expansions of T cells expressing homologous or identical complementary-determining region 3. Furthermore, T cell clones isolated from the two compartments expressed similar, sometimes identical, junctional regions. Removal from antigenic exposure led to the disappearance of T cell clones. Our findings indicate that expansions of T lymphocytes bearing clonal TCRBV region gene segments take place in the lung of patients with HP during exposure. The evidence that identical T cell clones are present in the lung and the blood of the same patient suggests that the immune reaction occurring at lung level gives rise to a systemic reaction.
Assuntos
Alveolite Alérgica Extrínseca/imunologia , Linfócitos T/imunologia , Adulto , Alveolite Alérgica Extrínseca/etiologia , Alveolite Alérgica Extrínseca/patologia , Sequência de Aminoácidos , Células Sanguíneas/imunologia , Estudos de Casos e Controles , Células Clonais/imunologia , Regiões Determinantes de Complementaridade/genética , Feminino , Genes Codificadores da Cadeia beta de Receptores de Linfócitos T , Humanos , Imunofenotipagem , Pulmão/imunologia , Pulmão/patologia , Ativação Linfocitária/imunologia , Masculino , Pessoa de Meia-Idade , Receptores de Antígenos de Linfócitos T/genéticaRESUMO
Glutathione S-Transferase enzymes (GSTs) constitute the principal Phase II superfamily which plays a key role in cellular detoxification and in other biological processes. Studies of GSTs have revealed that genetic polymorphisms are present in these enzymes and that some of these are Loss-of-Function (LoF) variants, which affect enzymatic functions and are related to different aspects of human health. The aim of this study was to analyze functional genetic differences in GST enzymes among human populations. Attention was focused on LoF polymorphisms of GSTA1, GSTM1, GSTO1, GSTO2, GSTP1 and GSTT1 genes. These LoF variants were analyzed in 668 individuals belonging to six human groups with different ethnic backgrounds: Amhara and Oromo from Ethiopia; Colorado and Cayapa Amerindians and African Ecuadorians from Ecuador; and one sample from central Italy. The HapMap database was used to compare our data with reference populations and to analyze the haplotype and Linkage Disequilibrium diversity in different ethnic groups. Our results highlighted that ethnicity strongly affects the genetic variability of GST enzymes. In particular, GST haplotypes/variants with functional impact showed significant differences in human populations, according to their ethnic background. These data underline that human populations have different structures in detoxification genes, suggesting that these ethnic differences influence disease risk or response to drugs and therefore have implications for genetic association studies involving GST enzymes. In conclusion, our investigation provides data about the distribution of important LoF variants in GST genes in human populations. This information may be useful for designing and interpreting genetic association studies.
Assuntos
Etnicidade/genética , Variação Genética , Glutationa Transferase/genética , Alelos , Frequência do Gene , Genética Populacional , Genótipo , Glutationa Transferase/metabolismo , HumanosAssuntos
Diabetes Mellitus Tipo 2/fisiopatologia , Retinopatia Diabética/patologia , Doenças Vasculares Periféricas/patologia , Células-Tronco/fisiologia , Adulto , Idoso , Antígenos CD34/análise , Diferenciação Celular/fisiologia , Retinopatia Diabética/imunologia , Retinopatia Diabética/fisiopatologia , Células Endoteliais/imunologia , Endotélio Vascular/citologia , Endotélio Vascular/fisiopatologia , Humanos , Pessoa de Meia-Idade , Neovascularização Patológica , Doenças Vasculares Periféricas/imunologia , Doenças Vasculares Periféricas/fisiopatologia , Estudos ProspectivosRESUMO
BACKGROUND: Circulating progenitor cells (PCs) may play a role in the pathogenesis of cardiac allograft vasculopathy, the leading cause of morbidity and mortality in heart transplantation (HTx). We assessed the relationship between circulating PCs and their incorporation into allografts and coronary microvascular function in HTx. METHODS: PCs were quantified by flow cytometry on the basis of the surface expression of CD34, CD133, and kinase domain receptor (KDR) antigens. Biopsy specimens at 2 different times were examined. Immunohistochemistry for the stem cell marker c-Kit, endothelial PC (EPC) marker KDR, and CD34 was performed in serial sections in all specimens. Cells positive for each marker were counted in all specimen area sections, and the number obtained was corrected by area section. Coronary flow in the left anterior descending coronary artery was detected at rest and during intravenous adenosine by transthoracic echocardiography. Coronary flow reserve (CFR) was the ratio of hyperemic diastolic mean velocity (DMV)/resting DMV. RESULTS: CFR was measured in 29 patients and was abnormal (CFR < 2) in 6 (Group A) and normal in 23 (Group B). CFR was lower in Group A (1.5 ± 0.1 vs 3.3 ± 0.8, p < 0.0001). CD34(+)KDR(+), CD133(+)KDR(+), and CD34(+)CD133(+)KDR(+) cell counts were lower in Group A (p < 0.05). EPCs in biopsy sections tended to be lower in Group A (p = 0.06) and correlated to circulating CD133(+)KDR(+) and CD34(+)CD133(+)KDR(+) (p = 0.003 and p = 0.052, respectively). CONCLUSIONS: EPCs are decreased in the circulation and in the allograft in patients with microvasculopathy. Defective mobilization and engraftment of EPCs may be involved in the pathogenesis of cardiac allograft vasculopathy.
Assuntos
Movimento Celular , Endotélio Vascular/patologia , Transplante de Coração/efeitos adversos , Células-Tronco/patologia , Doenças Vasculares/etiologia , Antígeno AC133 , Adulto , Antígenos CD/metabolismo , Antígenos CD34/metabolismo , Velocidade do Fluxo Sanguíneo , Contagem de Células , Circulação Coronária , Endotélio Vascular/metabolismo , Feminino , Glicoproteínas/metabolismo , Transplante de Coração/patologia , Humanos , Masculino , Microcirculação , Pessoa de Meia-Idade , Peptídeos/metabolismo , Células-Tronco/metabolismo , Transplante Homólogo , Doenças Vasculares/fisiopatologia , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismoRESUMO
PURPOSE: This study aimed to assess vascular endothelial function in patients with ocular hypertension (OHT) or primary open-angle glaucoma (POAG) by measuring: (a) endothelium-dependent flow-mediated vasodilation (FMD) of the brachial artery, and (b) circulating endothelial progenitor cells, which are believed to support the integrity of the vascular endothelium. METHODS: We enrolled 25 patients with OHT, 23 with POAG and 26 control subjects, all of whom were aged < 65 years and had no medical history of cardiovascular disease or cardiovascular risk factors. All subjects underwent a complete ophthalmological examination, biochemistry study, assessment of cardiovascular parameters, brachial artery ultrasound assessment of endothelium-dependent FMD, generic circulating progenitor cell (CPC) and circulating endothelial progenitor cell (EPC) count with the use of flow cytometry. RESULTS: Flow-mediated vasodilation values differed significantly in OHT (4.5 +/- 1.1%; p = 0.021) and POAG (4.0 +/- 0.9%; p = 0.003) patients compared with controls (7.7 +/- 0.8%). The CD34(+) KDR(+) EPC count was markedly lower in OHT (28.0 +/- 5.0; p < 0.001) and POAG (24.3 +/- 3.4; p < 0.001) patients compared with controls (73.1 +/- 8.1). Neither FMD not EPCs differed significantly between OHT and POAG patients. No significant differences in CPC count or cardiovascular parameters were found among OHT or POAG patients and controls. The levels of CD34(+) KDR(+) EPCs were directly correlated (p = 0.043) with FMD, and inversely correlated (p = 0.032) with baseline intraocular pressure in OHT and POAG patients. CONCLUSIONS: Both OHT and POAG patients without cardiovascular risk factors have previously unreported severely reduced circulating EPCs and reduced FMD, both of which are indicators of endothelial dysfunction and increased risk of cardiovascular events.
Assuntos
Artéria Braquial/fisiopatologia , Endotélio Vascular/patologia , Endotélio Vascular/fisiopatologia , Glaucoma de Ângulo Aberto/fisiopatologia , Hipertensão Ocular/fisiopatologia , Células-Tronco/patologia , Vasodilatação , Adulto , Contagem de Células , Feminino , Glaucoma de Ângulo Aberto/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Hipertensão Ocular/patologia , Fluxo Sanguíneo RegionalRESUMO
In the last 10 years an increasing interest has been devoted to the study of endothelial progenitor cells (EPCs), a subtype of immature cells involved in endothelial repair and neoangiogenesis. EPCs have been discovered as a novel integrated part of the cardiovascular system, which plays a comprehensive role in tissue homeostasis. Consistently, alterations and/or reduction of the circulating EPC pool have been associated with different manifestations of cardiovascular disorders and atherosclerosis. This is why, the extent of the EPC pool is now considered a mirror of vascular health, while EPC reduction has become a surrogate biomarker of cardiovascular risk and of the ongoing vascular damage. Unfortunately, the methods used to study EPCs still lack standardization, and this is significantly decelerating progress in the field. In this review, we focus on some aspects related to the two methods used to assess circulating EPCs: flow cytometry and cell culture. We uncover the many traps hidden in the choice of the right protocol, and suggest the best solutions on the basis of evidence and background theories.
Assuntos
Antígenos CD/classificação , Células Endoteliais/citologia , Células-Tronco/citologia , Aterosclerose/fisiopatologia , Biomarcadores/sangue , Técnicas de Cultura de Células , Células Endoteliais/fisiologia , Citometria de Fluxo/métodos , Humanos , Neovascularização Fisiológica/fisiologia , Kit de Reagentes para Diagnóstico , Células-Tronco/fisiologiaRESUMO
Chemokines and their receptors play a pivotal role in the regulation of B-lymphocyte trafficking. This study was aimed at investigating the pattern of chemokine receptor expression, including CCR1 to CCR3, CCR5 to CCR7, CXCR1 to CXCR5, and the migration ability of multiple myeloma (MM) plasma cells (PC). PC were recovered from the bone marrow (BM) of 29 MM patients, extramedullary sites of 10 patients and the BM of five controls. Flow cytometry analysis showed that the receptors mainly expressed on malignant BM PC were represented by CXCR4 (70% of patients), CCR1 (25%), CCR2 (25%), CCR5 (17%) and CXCR3 (20%), while other receptors were commonly lacking. The analysis performed on extramedullary (peripheral blood and pleural effusion) malignant PC demonstrated that the most represented receptors were CXCR4 (100%), CCR2 (66%) and CXCR1 (60%). The migratory capability of malignant PC at resting conditions identified three groups of patients with different migration (low, intermediate and high). As CXCR4 was the relevant chemokine receptor expressed by MM PC, its ligand CXCL12 induced their migration. These data suggest that malignant PC from MM display different chemokine receptor profiles and that CXCR4 is fully functional and might play a role in the spreading of the disease.
Assuntos
Mieloma Múltiplo/imunologia , Plasmócitos/imunologia , Receptores de Quimiocinas/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Células da Medula Óssea/imunologia , Quimiocina CXCL12 , Quimiocinas CXC/metabolismo , Quimiotaxia de Leucócito/imunologia , Feminino , Citometria de Fluxo , Humanos , Masculino , Pessoa de Meia-Idade , Derrame Pleural/imunologia , Receptores CCR2 , Receptores CXCR4/metabolismo , Receptores de Interleucina-8A/metabolismo , Células Tumorais CultivadasRESUMO
AIMS: Circulating progenitor cells are believed to participate in cardiovascular (CV) homeostasis and their exhaustion has been linked to CV damage. As general agreement on their characterization is lacking, this work was carried out to assess the relationships between different antigenic profiles of progenitor cells and CV risk, with special regard to metabolic syndrome (MetSyn). METHODS AND RESULTS: CD34, CD133, and KDR were used to quantify circulating progenitors in 214 subjects at different levels of CV risk. In a cross-analysis of six different cell subtypes (CD34(+), CD133(+), CD34(+)CD133(+), CD34(+)KDR(+), CD133(+)KDR(+), and CD34(+)CD133(+)KDR(+)), CD34(+) progenitors showed the best correlation with CV parameters and risk estimates. Components of the MetSyn were all characterized by reduction of CD34(+) cells and acted synergistically in decreasing CD34(+) cell count. Moreover, CD34(+) cell count demonstrated a high performance in detecting high CV risk. CONCLUSION: These data demonstrate that CD34 identifies progenitor cells linked to CV risk, showing a close negative correlation between CD34(+) cells and CV risk, as well as a synergic detrimental effect of clustered metabolic components. Progenitor cell count may be used as a surrogate marker of CV risk, whereas extensive antigenic characterization may not be useful for this purpose.