RESUMO
Inositol 1,4,5-triphosphate (IP3) and inositol 1,3,4,5-tetrakisphosphate (IP4) are calcium-regulating second messenger molecules generated following the binding of a wide range of hormones and growth factors to their receptors. The actions of these messengers, which play important roles in the regulation of cell proliferation as well as in other signaling pathways, are terminated by the action of a 5-phosphomonoesterase (5-PME) enzyme. We have assayed this enzyme in normal and malignant hemopoietic cells. Extracts from normal bone marrow cells and peripheral blood mononuclear cells (PBMNC) degraded [3H]IP3 at rates of 74.5 (+/- 3.4) and 84.5 (+/- 7.9) pmol/min/micrograms protein, respectively. PME activity in 10/13 (77%) acute lymphoblastic leukemia samples were significantly below the normal range and the enzyme was completely undetectable in three (23%) of these. Enzyme activity in 8/9 (89%) chronic lymphocytic leukemia samples were below the normal range, being undetectable in three of these (33%). Nine of 24 (38%) acute myeloid leukemia samples contained low 5-PME levels, which was undetectable in one sample. Reduced 5-PME activity was detected in 2/7 (28%) of chronic granulocytic leukemia samples. The data here are consistent with the hypothesis that a reduced rate of degradation of IP3 and IP4 in some leukemia cells may result in the aberrant operation of signaling pathways, possibly including those involved in the control of cell proliferation.
Assuntos
Cálcio/metabolismo , Fosfatos de Inositol/metabolismo , Leucemia/metabolismo , Monoéster Fosfórico Hidrolases/metabolismo , Sistemas do Segundo Mensageiro , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Medula Óssea/enzimologia , Criança , Pré-Escolar , Feminino , Células-Tronco Hematopoéticas/enzimologia , Humanos , Immunoblotting , Inositol 1,4,5-Trifosfato/metabolismo , Leucemia/enzimologia , Leucemia/patologia , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/enzimologia , Lítio/farmacologia , Masculino , Pessoa de Meia-Idade , Testes de PrecipitinaRESUMO
Inositol 1,4,5-trisphosphate and inositol 1,3,4,5-tetrakisphosphate are Ca(2+)-regulating second messenger molecules which are generated via the cleavage of inositol lipids. We have previously shown that these species are autonomously generated in HL60 myeloid leukemia cells and that they may play a role in signalling the continuous proliferation of this cell line. Here we show that the activity of the 5-phosphomonoesterase (5-PME) enzyme which cleaves and inactivates these second messengers was strikingly reduced in HL60 cells compared to normal granulocytes or macrophages. Induction of differentiation of HL60 cells along the monocyte/macrophage or granulocytic pathways did not result in a significant increase in 5-PME activity. The activity of this enzyme was also low in extracts of bone marrow mononuclear cells from four patients with myeloid leukemia. A lesion in the 5-PME pathway may therefore result in the conservation of Ca(2+)-regulating second messengers in the HL60 cell line and in some myeloid leukemia cells. It is plausible that this lesion may co-operate with the autonomous cleavage of inositol lipids in the signalling of leukemic cell proliferation.
Assuntos
Inositol 1,4,5-Trifosfato/metabolismo , Leucemia Mieloide/metabolismo , Medula Óssea/metabolismo , Cálcio/metabolismo , Divisão Celular/fisiologia , Fracionamento Celular , Humanos , Leucemia Mieloide/patologia , Sistemas do Segundo Mensageiro , Trítio , Células Tumorais CultivadasRESUMO
Infection with HIV causes a reduction in the numbers and function of CD4+ lymphocytes and functional abnormalities of other cells. We have studied the effect of HIV infection on signal transduction in the H9 lymphoblastoid CD4+ cell line. Resting HIV-infected H9 cells show evidence of chronic activation with raised levels of InsP3 and InsP4, the active metabolites of the inositol polyphosphate pathway, and a consequently raised intracellular free calcium concentration. Stimulation of HIV-infected H9 cells with phytohemagglutinin (PHA) leads to a fall in the previously raised levels of InsP3 but a further rise in InsP4, whilst an attenuated intracellular calcium rise is seen with both PHA and anti-CD3 antibody. The observed effects of HIV infection on signal transduction provide a mechanism to explain the functional defects in CD4+ lymphocytes and, possibly, other cell types.
Assuntos
Linfócitos T CD4-Positivos/microbiologia , Fosfatos de Inositol/metabolismo , Linfócitos T CD4-Positivos/metabolismo , Cálcio/metabolismo , Linhagem Celular , Humanos , Hidrólise , Fito-Hemaglutininas/farmacologia , Transdução de SinaisRESUMO
Lymphocytes or lymphoblastoid cells that have been infected by HIV in vitro or exposed to its envelope glycoprotein (gp120) show abnormal inositol polyphosphate-mediated signal transduction and associated defects in calcium regulation. Such cells behave as though they were chronically activated and fail to respond to further activating signals. We now show that similar changes are seen in lymphocytes obtained from HIV-infected subjects at various stages of infection, despite the fact that only a minority of such cells are infected. Furthermore, the defect in the phosphatidylinositol hydrolysis pathway in lymphocytes obtained from AIDS patients reverses after treatment with zidovudine, in parallel with improvements in phytohaemagglutinin-induced proliferative response and interferon-gamma production.
Assuntos
Síndrome da Imunodeficiência Adquirida/sangue , Infecções por HIV/sangue , Fosfatos de Inositol/metabolismo , Linfócitos/metabolismo , Zidovudina/uso terapêutico , Síndrome da Imunodeficiência Adquirida/tratamento farmacológico , Linfócitos T CD4-Positivos , Cálcio/metabolismo , Infecções por HIV/tratamento farmacológico , Humanos , Inositol 1,4,5-Trifosfato/metabolismo , Interferon gama/metabolismo , Contagem de Leucócitos , Ativação Linfocitária/efeitos dos fármacos , Linfócitos/efeitos dos fármacos , Fosfatidilinositóis/metabolismo , Linfócitos T ReguladoresRESUMO
OBJECTIVES: To investigate, in lymphocytes from HIV-1-infected individuals, the phenotypic expression of various adhesion co- or counter-receptors [lymphocyte function-associated antigen (LFA)-3, LFA-1 and intercellular adhesion molecule (ICAM)-1] involved in providing the co-stimulatory signal through the phospholipase C-gamma pathway in relation to inositol polyphosphate metabolism. DESIGN AND METHODS: Cell adhesion molecule profiles of peripheral blood lymphocytes (PBL) from 39 HIV-1-infected individuals at various stages of infection and 20 healthy laboratory controls were studied using flow cytometry. These were studied in 14 patients with late-stage disease in conjunction with their inositol polyphosphate metabolic profiles measured by high performance liquid chromatography. Levels of HIV-1 present in cell lysates were concurrently measured by a p24 antigen capture assay. In addition, the effects of a specific anti-ICAM-1 antisense oligonucleotide on the intracellular phosphatase activities of lymphocytes from a separate group of eight HIV-1-infected individuals were examined. RESULTS: The expression of LFA-1, a beta 2 integrin, was upregulated among patient PBL in parallel with disease progression, whereas that of LFA-3 (CD58) was found to be significantly reduced among the CD4+ lymphocyte subset in all stages of infection. The 5-phosphatase activity, which we previously observed to be defective in HIV disease, was found to correlate linearly with the expression of both LFA-1 and its ligand, ICAM-1. Treatment of patient lymphocytes with an antisense oligonucleotide, which reduced the cell surface expression of ICAM-1 by blocking the translation of its mRNA, resulted in further reduction of intracellular phosphatase activities. CONCLUSIONS: Our results suggest a pivotal role for adhesion co- and counter-receptors in influencing lymphocyte signalling and hence cellular response to recall antigens in HIV-1-infected individuals.
Assuntos
Moléculas de Adesão Celular/metabolismo , Infecções por HIV/imunologia , Infecções por HIV/terapia , HIV-1 , Antígenos CD/metabolismo , Sequência de Bases , Antígenos CD58 , Infecções por HIV/metabolismo , Humanos , Imunoterapia , Técnicas In Vitro , Fosfatos de Inositol/metabolismo , Inositol Polifosfato 5-Fosfatases , Molécula 1 de Adesão Intercelular/genética , Molécula 1 de Adesão Intercelular/metabolismo , Antígeno-1 Associado à Função Linfocitária/metabolismo , Linfócitos/efeitos dos fármacos , Linfócitos/imunologia , Linfócitos/metabolismo , Masculino , Glicoproteínas de Membrana/metabolismo , Dados de Sequência Molecular , Oligonucleotídeos Antissenso/genética , Oligonucleotídeos Antissenso/farmacologia , Monoéster Fosfórico Hidrolases/metabolismo , Transdução de SinaisRESUMO
Although circulating levels of interleukin 8 (IL-8), a potent pro-inflammatory chemokine, and many other inflammatory mediators increase in response to cardiopulmonary bypass, only a small proportion of patients develop a clinically significant systemic inflammatory response. The natural mechanisms that control the inflammatory response are poorly understood. To investigate the role of IL-8 in a human inflammatory model, 15 adult patients undergoing cardiopulmonary bypass for elective coronary artery bypass grafting were studied. Following reperfusion, plasma IL-8 levels increased significantly from 58 pg/mL (pre-bypass) and 66 pg/mL (after 20 min of bypass) to 98 pg/mL (p = .02 and .04, respectively), but this was accompanied by a concomitant threefold decrease in the IL-8 binding affinity of circulating neutrophils (Dissociation constant (KL) post-reperfusion/KL pre-bypass = 3.2; KL post-reperfusion/KL after 20 min of bypass = 2.8). IL-8-triggered release of myeloperoxidase and elastase by peripheral blood neutrophils ex vivo was also down-regulated following reperfusion. There were no significant changes in beta 2 integrin expression or inositol polyphosphate metabolism of peripheral blood neutrophils. These changes in receptor affinity and neutrophil responsiveness to IL-8 may represent an important in vivo regulatory mechanism which serves to prevent excessive tissue injury from inflammatory triggers.
Assuntos
Ponte Cardiopulmonar/efeitos adversos , Inflamação/fisiopatologia , Neutrófilos/metabolismo , Antígenos CD/metabolismo , Antígenos CD18/metabolismo , Reagentes de Ligações Cruzadas , Humanos , Fosfatos de Inositol/metabolismo , Interleucina-8/sangue , Interleucina-8/metabolismo , Interleucina-8/farmacologia , Elastase de Leucócito/metabolismo , Neutrófilos/efeitos dos fármacos , Peroxidase/metabolismo , Receptores de Interleucina/metabolismo , Receptores de Interleucina-8A , Transdução de SinaisRESUMO
Qualitative defects in immune responsiveness after human immunodeficiency virus (HIV) infection have been well characterized and may play a key role in the development of HIV disease. However, no clear picture of the underlying mechanism of the functional deficiencies has yet emerged. In this article, Anthony Pinching and Keith Nye suggest that HIV or HIV proteins can sabotage transmembrane signalling and that this is of primary importance to the alterations in immune responsiveness.
Assuntos
Infecções por HIV/fisiopatologia , HIV/fisiologia , Modelos Biológicos , Transdução de Sinais , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/fisiologia , Cálcio/metabolismo , Proteína gp120 do Envelope de HIV/fisiologia , Infecções por HIV/imunologia , Humanos , Tolerância Imunológica , Fosfatos de Inositol/metabolismo , Ativação Linfocitária , Macrófagos/fisiologia , Neurônios/fisiologia , Especificidade de ÓrgãosRESUMO
Lymphocytes infected in vivo with HIV or lymphoblastoid cells exposed in vitro to either HIV or its envelope glycoprotein (gp120) show a defect in inositol polyphosphate-mediated signal transduction together with an associated abnormality in intracellular calcium regulation. The defect in patients reverses after treatment with the anti-retroviral agent zidovudine (AZT). We present evidence that the defect is at the level of the Ins (1,3,4,5)P4 5-phosphomonoesterase (PME) in these cells and that, though elevation of the intracellular ATP level partially down-regulates the activity of this enzyme, such changes alone are unable to account for the complete inhibition seen in HIV-infected cells.
Assuntos
HIV/metabolismo , Linfócitos/metabolismo , Fosfatidilinositóis/metabolismo , Monoéster Fosfórico Hidrolases/fisiologia , Trifosfato de Adenosina/farmacologia , Cálcio/farmacologia , Cromatografia Líquida de Alta Pressão , Relação Dose-Resposta a Droga , Regulação para Baixo/efeitos dos fármacos , Proteína gp120 do Envelope de HIV/imunologia , Humanos , Hidrólise , Inositol Polifosfato 5-Fosfatases , Transdução de Sinais , Zidovudina/farmacologiaRESUMO
The effect of early human immunodeficiency virus-1 infection in vitro on proximal signal transduction events in primary peripheral blood lymphocytes was investigated with respect to CD4-mediated costimulation of CD3/T cell-receptor signalling. Tyrosine phosphorylation profiles induced by CD4 and CD3 + CD4 ligation were profoundly abrogated in virally infected cells, although CD4 receptor expression remained intact during early infection. Furthermore, the association of the tyrosine kinase p56lck with the CD4 receptor was reduced in virally infected cells. The downmodulation of CD4-mediated CD3 signalling coincided with the subsequent inhibition of the activity and tyrosine phosphorylation of the downstream kinase ZAP-70 in virally infected cells. The observed virally mediated cosignalling defects during early infection may account for the inhibition of distal signal events and thus contribute to HIV pathogenesis, such as reduced immune response to antigenic exposure, anergy, and apoptosis.
Assuntos
Antígenos CD4/metabolismo , Linfócitos T CD4-Positivos/virologia , HIV-1/crescimento & desenvolvimento , Proteínas Tirosina Quinases/metabolismo , Quinases da Família src/metabolismo , Complexo CD3/metabolismo , Células Cultivadas , Regulação para Baixo , Humanos , Capeamento Imunológico , Proteína Tirosina Quinase p56(lck) Linfócito-Específica , Fosforilação , Transdução de Sinais , Proteína-Tirosina Quinase ZAP-70RESUMO
We investigated the effects of early human immunodeficiency virus-1 infection (HIV-1) on CD4- and CD28-mediated co-signaling of the T cell receptor (TCR)/CD3 complex in peripheral blood lymphocytes (PBL). CD4 ligation either alone or in conjunction with TCR occupancy resulted in abrogated signaling shown by impaired co-association of the tyrosine kinase ZAP-70 with the CD3-zeta chains in virally infected PBL. In addition, down-regulation of CD4-associated TCR signaling resulted in diminished tyrosine phosphorylation of mitogen-activated protein kinase (MAPK), a serine threonine kinase which is critically involved in the regulation of transcription factors. Furthermore, these aberrant CD4-driven signals rendered HIV-1-infected PBL susceptible to activation-induced cell death. By contrast, cross-linking of the TCR/CD3 complex with the CD28 receptor improved tyrosine phosphorylation of MAPK and salvaged infected PBL from activation-induced cell death. Our data demonstrate the importance of appropriate CD3, CD4 and CD28 co-stimulatory function to prevent apoptosis. The CD4-mediated signaling defects of the TCR could contribute to the loss of immunocompetent cells during HIV-1 infection via activation-induced cell death, whereas stimulation through the CD28 pathway could reverse these detrimental effects.
Assuntos
Infecções por HIV/imunologia , HIV-1 , Linfócitos/imunologia , Apoptose/imunologia , Antígenos CD28/metabolismo , Complexo CD3/metabolismo , Antígenos CD4/metabolismo , Proteínas Quinases Dependentes de Cálcio-Calmodulina/metabolismo , Sobrevivência Celular , Infecções por HIV/metabolismo , Infecções por HIV/patologia , Humanos , Técnicas In Vitro , Linfócitos/metabolismo , Linfócitos/patologia , Fosforilação , Proteínas Tirosina Quinases/metabolismo , Receptores de Antígenos de Linfócitos T/metabolismo , Transdução de Sinais/imunologia , Tirosina/metabolismo , Proteína-Tirosina Quinase ZAP-70RESUMO
Proper function of the protein tyrosine phosphatase CD45 is required for the positive regulation of the activity of src tyrosine kinases p56lck and p59fyn which participate in T-cell receptor and CD4 receptor signalling. In this study, the effect of HIV-1 infection on the function of CD45-associated tyrosine phosphatase activity in the H9 T-cell line has been investigated with respect to CD3 and CD4 ligation. A significant reduction in CD45-associated phosphatase activity was observed following CD3 + CD4 ligation in virally infected cells, whereas CD45 activity was not compromised following CD3 receptor ligation. Dysfunctional CD45 activity in infected cells was not attributable to reduced receptor surface expression induced by HIV-1, since CD4, CD3 and CD45 expression levels were found to be intact. Defective CD45 activity correlated with inhibted downstream signalling events as evidenced by reduced CD4-associated tyrosine kinase activity and inhibition of PLC-gamma1. Impaired CD45 function is likely to play a critical role in the inhibition of CD3/CD4 signalling thereby contributing to HIV-1 pathogenesis.
Assuntos
Complexo CD3/fisiologia , Antígenos CD4/fisiologia , HIV-1/fisiologia , Antígenos Comuns de Leucócito/fisiologia , Proteínas Tirosina Fosfatases/metabolismo , Transdução de Sinais , Sequência de Aminoácidos , Linhagem Celular , Meios de Cultura Livres de Soro , Gastrinas/química , Gastrinas/metabolismo , Proteína do Núcleo p24 do HIV/análise , HIV-1/imunologia , Humanos , Dados de Sequência Molecular , Proteínas Tirosina Quinases/metabolismo , Receptor Cross-Talk , Especificidade por SubstratoRESUMO
The human immunodeficiency virus-1 (HIV-1) utilises CD4 and certain beta-chemokine receptors, mainly CCR-5 and CXCR4, for attachment and virus entry into T-lymphocytes and monocytes/macrophages. CD4 and beta-chemokine receptors participate in intracellular signalling via protein tyrosine kinases and G-protein-coupled signalling. The factors which influence HIV-1 replication and the intracellular signalling mechanisms elicited by the virus are not well understood. In this study, it was demonstrated that exposure of peripheral blood lymphocytes (PBLs) to a T-cell tropic strain of HIV-1 evokes signal(s) which results in downregulation of intracellular cAMP. In addition, pre-incubation of PBLs with the Gi-protein inhibitor Pertussis toxin mediated a significant inhibition of HIV-1 replication. These data strongly suggest that HIV-1 employs CD4 receptors and Gi-coupled proteins for entry into target cells and that productive HIV-1 infection is dependent on an active signalling event.
Assuntos
AMP Cíclico/metabolismo , Proteínas de Ligação ao GTP/fisiologia , HIV-1/fisiologia , Linfócitos/virologia , Toxina Pertussis , Transdução de Sinais , Fatores de Virulência de Bordetella/farmacologia , Benzoquinonas , Antígenos CD4/metabolismo , Divisão Celular , Sobrevivência Celular/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , Citometria de Fluxo , Imunofluorescência , Subunidades alfa Gi-Go de Proteínas de Ligação ao GTP/antagonistas & inibidores , Subunidades alfa Gi-Go de Proteínas de Ligação ao GTP/fisiologia , Proteínas de Ligação ao GTP/antagonistas & inibidores , Proteína do Núcleo p24 do HIV/metabolismo , HIV-1/patogenicidade , Humanos , Lactamas Macrocíclicas , Linfócitos/citologia , Proteínas Tirosina Quinases/antagonistas & inibidores , Proteínas Tirosina Quinases/metabolismo , Quinonas/farmacologia , Receptores CXCR4/metabolismo , Rifabutina/análogos & derivados , Transdução de Sinais/efeitos dos fármacos , Fatores de Tempo , Replicação Viral/efeitos dos fármacosRESUMO
The distribution of phenotypes of the group specific component (Gc) was examined in 203 homosexuals at risk of infection or infected by the human immunodeficiency virus and compared with that in 50 randomly selected homosexuals and 122 healthy male heterosexual seronegative controls. 30.2% of patients with the acquired immunodeficiency syndrome (AIDS) were homozygous for Gc 1 fast (Gc 1f) compared with 0.8% of controls (p less than 0.0001); patients with other clinical manifestations of HIV infection were also more likely than controls to have Gc 1f. By contrast, seronegative symptomless homosexual contacts of AIDS patients (AH-p) lacked this phenotype but were more likely than controls to be homozygous for Gc 2 (25% vs 9%, p less than 0.05). AIDS patients lacked the homozygous Gc 2 phenotype altogether. A chi 2 trend test showed that progression to AIDS had a strong positive association with the Gc 1f allele (p less than 0.0001) and a negative one with Gc 2 (p less than 0.05). It is proposed that Gc may be involved in viral entry into host cells, the ease of which varies with different allelic forms of Gc, according to their sialic acid content.
Assuntos
Síndrome da Imunodeficiência Adquirida/genética , Alelos , HIV/genética , Proteína de Ligação a Vitamina D/análise , Complexo Relacionado com a AIDS/genética , Anticorpos Antivirais/análise , Suscetibilidade a Doenças , Eletroforese em Gel de Poliacrilamida , Anticorpos Anti-HIV , Homossexualidade , Homozigoto , Humanos , Masculino , Fenótipo , Risco , Comportamento Sexual , Proteína de Ligação a Vitamina D/genéticaRESUMO
Rheumatic fever is associated with exaggerated activity of B cells with massive production of antibody to the Group A streptococcus. Gc (vitamin D-binding protein) is constitutively expressed on B-cell membranes in association with membrane immunoglobulin, and could be involved in cell activation. We therefore looked for associations between the three major Gc alleles and susceptibility to rheumatic fever in a homogeneous Arab population. Patients with tuberculosis or rheumatoid arthritis and control donors, were studied in parallel. Allele frequencies in the controls, rheumatoid and tuberculosis patients were identical to those found in a previous study of normal Arab donors. However, there was a striking association between Gc2 and rheumatic fever. This allele was twice as common in these patients as in controls (p = 0.0024), and was present in 56.4% of all rheumatic fever patients.
Assuntos
Alelos , Febre Reumática/genética , Proteína de Ligação a Vitamina D/genética , Criança , Suscetibilidade a Doenças , Feminino , Humanos , MasculinoRESUMO
HIV infection is associated with a disease status-dependent impairment of Ag-specific T cell responses, resulting in anergy or unchecked apoptotic cell death. beta1 integrins play an important role in the induction of T lymphocyte responses to antigenic challenge by providing a T cell costimulatory signal, and have been shown to rescue various cell types from undergoing apoptosis. We examined the integrin-triggered cell survival signal and associated pathways in CD3+ T cells derived from 69 HIV-1-infected individuals in comparison with healthy controls. We found beta1 integrin-mediated costimulation of TCR-induced T cell proliferation and protection from aberrant cell death to be absent in the majority of patients with AIDS, but intact in asymptomatic, infected individuals. The lack of integrin-mediated rescue may be partly due to an early impairment of TCR/integrin-costimulated secretion of IFN-gamma, a type 1 lymphokine that protects against TCR-induced apoptosis of T cells from HIV-seropositive donors, but not loss of integrin expression. The mechanism of integrin hyporesponsiveness appeared to correlate with a failure of the integrin-generated signal to induce pp125FAK mRNA and protein expression. Protein kinase C activation in CD3+ T cells following integrin stimulation was also impaired in HIV-infected individuals, mostly among the symptomatic/AIDS patients. Protein kinase C inactivation in T cells was shown to have a destabilizing effect in vitro on pp125FAK mRNA that contains an AUUUA motif in the 3'-untranslated region, a consensus sequence for the AU-rich elements responsible for mRNA destabilization. These aberrant changes in pp125FAK expression may have direct significance to the overall immunopathogenesis during infection with HIV-1.
Assuntos
Apoptose/imunologia , Infecções por HIV/imunologia , Integrinas/fisiologia , Linfócitos T/imunologia , Síndrome da Imunodeficiência Adquirida/imunologia , Fármacos Anti-HIV/uso terapêutico , Anticorpos Monoclonais/farmacologia , Apoptose/efeitos dos fármacos , Linfócitos T CD4-Positivos/imunologia , Moléculas de Adesão Celular/biossíntese , Moléculas de Adesão Celular/sangue , Moléculas de Adesão Celular/genética , Sinergismo Farmacológico , Ativação Enzimática/efeitos dos fármacos , Epitopos/fisiologia , Quinase 1 de Adesão Focal , Proteína-Tirosina Quinases de Adesão Focal , Infecções por HIV/tratamento farmacológico , Infecções por HIV/metabolismo , Humanos , Tolerância Imunológica , Integrina beta1/biossíntese , Integrinas/metabolismo , Interferon gama/metabolismo , Interfase , Leucemia Linfoide , Ativação Linfocitária/efeitos dos fármacos , Proteína Quinase C/efeitos dos fármacos , Proteínas Tirosina Quinases/biossíntese , Proteínas Tirosina Quinases/sangue , Proteínas Tirosina Quinases/genética , RNA Mensageiro/biossíntese , Complexo Receptor-CD3 de Antígeno de Linfócitos T/antagonistas & inibidores , Complexo Receptor-CD3 de Antígeno de Linfócitos T/biossíntese , Complexo Receptor-CD3 de Antígeno de Linfócitos T/fisiologia , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/imunologia , Linfócitos T/efeitos dos fármacos , Células Tumorais CultivadasRESUMO
OBJECTIVE: To establish the relationship between T cell responses to integrin coreceptor stimulation and B cell hyperreactivity as measured by pathologic autoantibody production. METHODS: Peripheral blood mononuclear cells from 42 patients with SLE according to the American Rheumatism Association criteria were examined for their ability to adhere to plate-immobilised fibronectin. Co-stimulation assays were performed on the same cells using anti-CD3 antibody alone or co-immobilised with an anti-beta1-integrin antibody. Proliferative responses were measured by 3[H]thymidine pulsing on day 3 and activation was determined using a commercial protein kinase C assay, the protocol being established by our group in association with Promega. Beta-integrin expression was established by FACS analysis. RESULTS: An impaired PKC response to integrin-mediated activation was found in T-lymphocytes from 6/21 (29%) SLE patients, which correlated significantly with an absence of anti-dsDNA antibody in patient sera, irrespective of prednisolone treatment. Integrin co-stimulation of TcR/CD3-induced proliferation and T cell adhesion to fibronectin were also impaired among 5/21 (24%) and 6/15 (40%) patients studied, respectively. CONCLUSION: We hypothesise that the integrity of beta1-integrin signalling pathways may influence pathological antibody production in SLE by affecting T-lymphocyte activation and interactions between T- and B-lymphocytes.