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1.
Mucosal Immunol ; 12(6): 1358-1369, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31582819

RESUMO

De-regulated T-cell activation and functions are pivotal in the orchestration of immune-mediated tissue damage in IBD. We investigated the role of DNAM-1 (co-activating)/TIGIT (co-inhibitory)/ligand axis in the regulation of T-cell functions and its involvement in IBD pathogenesis. We show that DNAM-1 and TIGIT display a peculiar expression pattern on gut mucosa T-cell populations, in a microenvironment where their shared ligands (PVR and Nectin-2) are physiologically present. Moreover, DNAM-1 family receptor/ligand system is perturbed in IBD lesions, in a disease activity-dependent manner. The expression profile of CCR6 and CD103 mucosa addressins suggests that microenvironment-associated factors, rather than skewed recruitment of circulating T-cell populations, play a more relevant role in supporting the establishment of DNAM-1 and TIGIT expression pattern in mucosal T-cell populations, and may explain its alteration in IBD. Although both co-receptors mark functionally competent T cells, DNAM-1 and TIGIT segregate on T cells endowed with different proliferative potential. Moreover, their opposing role in regulating T-cell proliferation exquisitely depends on ligand availability. All together, our data propose a role for DNAM-1 and TIGIT in regulating mucosal T-cell activation and immune homeostasis, and highlight the involvement of an imbalance of this system in IBD.


Assuntos
Antígenos de Diferenciação de Linfócitos T/metabolismo , Colo/metabolismo , Doenças Inflamatórias Intestinais/metabolismo , Mucosa Intestinal/metabolismo , Ativação Linfocitária , Receptores Imunológicos/metabolismo , Linfócitos T/metabolismo , Adolescente , Fatores Etários , Estudos de Casos e Controles , Proliferação de Células , Microambiente Celular , Criança , Pré-Escolar , Colo/imunologia , Feminino , Células HT29 , Humanos , Imunidade nas Mucosas , Lactente , Doenças Inflamatórias Intestinais/imunologia , Mucosa Intestinal/imunologia , Masculino , Nectinas/metabolismo , Receptores Virais/metabolismo , Transdução de Sinais , Linfócitos T/imunologia
2.
Artigo em Inglês | MEDLINE | ID: mdl-30828351

RESUMO

BACKGROUND: Identifying the mechanisms responsible for the development of food allergy in liver transplant recipients is more complex as there are several different clinical scenarios related to the immunological function of the liver. CASE PRESENTATION: We describe the first case of Transplant Acquired Food Allergy (TAFA) to cow milk in an adult following LT from a donor dead because of anaphylactic shock. A 67-year-old woman with primary biliary cirrhosis was referred to the Transplant Center of our hospital because of an acute-on-chronic liver failure. The donor was a 15-year-old girl deceased for anoxic encephalopathy due to food induced anaphylaxis after eating a biscuit. In the donor's history food allergies to cow milk and eggs were present. CONCLUSION: This case emphasizes the need for a standardized assessment of both solid-organ donors and recipients including donor allergy history in order to detect recipients at risk for anaphylaxis due to passive IgE transfer. Despite several reports of TAFA after solid organ, especially liver, an appropriate protocol to avoid risk for the recipient doesn't exist at the moment. The SPT (skin prick test) or specific IgE level are not enough to ensure a correct management in these cases and a correct education of the patients and the medical staff involved is absolutely necessary. It is the first case of milk allergy sensitization after solid organ transplant by passive transfer of IgE.

3.
Allergol. immunopatol ; 45(5): 452-456, sept.-oct. 2017. tab, graf
Artigo em Inglês | IBECS | ID: ibc-166999

RESUMO

Background: The diagnostic and therapeutic approach to grass pollen allergy is now possible by detecting specific IgE (sIgE) to its allergenic components. Aim: To evaluate the correlation between the sensitisation to different molecular Phleum pratense (Phl p) allergens and clinical efficacy of SLIT. Methods: The pilot study included 36 patients affected by allergic rhinoconjunctivitis, all treated with SLIT actively. We performed serum analysis of sIgE to Phl p 1, 2, 4, 5, 6, 7, 11 and 12. The Average Rhinoconjunctivitis Total Symptom Score (ARTSS) and the Average Combined Score (ACS) were evaluated before and after one year of immunotherapy. Results: Three different groups of sensitisation were defined based on the range of IgE reactivity to Phleum pratense allergens at baseline: group I (sIgE reactive to 1-3 allergens); group II (sIgE reactive to 4-5 allergens); and group III (sIgE reactive to 6-8 allergens). At T0 ACS was 1.79 ± 0.18 in group I; 1.81 ± 0.23 in group II; and 1.95 ± 0.34 in group III. At T1 ACS was 0.85 ± 0.55 in group I; 1.01 ± 0.31 in group II; and 1.44 ± 0.39 in group III. At T1 there was a significant improvement of ARTSS and ACS for group I (p = 0.001). Conclusions: Sublingual immunotherapy with a grass pollen is efficacious irrespective of the patients’ baseline sensitisation to either single or multiple grass pollen molecular allergens. We found that patients with few sensitisations have a greater improvement in combined symptom and medication score. SLIT improves the clinical course of allergic patients although new sensitisations may appear (AU)


No disponible


Assuntos
Humanos , Masculino , Feminino , Criança , Adolescente , Rinite Alérgica/terapia , Conjuntivite Alérgica/terapia , Imunoterapia Sublingual/métodos , Resultado do Tratamento , Pólen/efeitos adversos , Alérgenos/uso terapêutico
4.
Allergol Immunopathol (Madr) ; 45(5): 452-456, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28410874

RESUMO

BACKGROUND: The diagnostic and therapeutic approach to grass pollen allergy is now possible by detecting specific IgE (sIgE) to its allergenic components. AIM: To evaluate the correlation between the sensitisation to different molecular Phleum pratense (Phl p) allergens and clinical efficacy of SLIT. METHODS: The pilot study included 36 patients affected by allergic rhinoconjunctivitis, all treated with SLIT actively. We performed serum analysis of sIgE to Phl p 1, 2, 4, 5, 6, 7, 11 and 12. The Average Rhinoconjunctivitis Total Symptom Score (ARTSS) and the Average Combined Score (ACS) were evaluated before and after one year of immunotherapy. RESULTS: Three different groups of sensitisation were defined based on the range of IgE reactivity to Phleum pratense allergens at baseline: group I (sIgE reactive to 1-3 allergens); group II (sIgE reactive to 4-5 allergens); and group III (sIgE reactive to 6-8 allergens). At T0 ACS was 1.79±0.18 in group I; 1.81±0.23 in group II; and 1.95±0.34 in group III. At T1 ACS was 0.85±0.55 in group I; 1.01±0.31 in group II; and 1.44±0.39 in group III. At T1 there was a significant improvement of ARTSS and ACS for group I (p=0.001). CONCLUSIONS: Sublingual immunotherapy with a grass pollen is efficacious irrespective of the patients' baseline sensitisation to either single or multiple grass pollen molecular allergens. We found that patients with few sensitisations have a greater improvement in combined symptom and medication score. SLIT improves the clinical course of allergic patients although new sensitisations may appear.


Assuntos
Alérgenos/uso terapêutico , Antígenos de Plantas/uso terapêutico , Conjuntivite/terapia , Rinite Alérgica/terapia , Imunoterapia Sublingual/métodos , Adolescente , Alérgenos/imunologia , Antígenos de Plantas/imunologia , Criança , Conjuntivite/diagnóstico , Conjuntivite/imunologia , Feminino , Humanos , Imunização , Imunoglobulina E/sangue , Masculino , Phleum , Pólen , Valor Preditivo dos Testes , Rinite Alérgica/diagnóstico , Rinite Alérgica/imunologia
5.
Eur Rev Med Pharmacol Sci ; 19(1): 149-53, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25635988

RESUMO

Acute, drug-induced angioedema may not respond to standard therapies, because the pathogenetic mechanism that induces the pathology is not always mediated by histamine but, in certain instances, by bradykinin. A case of angioedema is reported here, in which allergic etiology was excluded by the non-response to antihistamines. Considering the clinical history (repeated use of drugs) and the ineffectiveness of standard therapy, it was decided to administer a beta2 receptor antagonist, icatibant. After 20 minutes, the patient reported a subjective improvement. The only form of angioedema for which this type of medication is licensed is the hereditary deficiency of C1 inhibitor. The use of icatibant for the treatment of other types of angioedema (which can also be life-saving if the airway is involved) is off label. The off-label use of a drug is allowed in the absence of a viable alternative therapy, if there is scientific evidence in the literature and if the prescriber takes responsibility. The case here reported draws attention to this therapeutic problem and underlines the fact that a life-threatening emergency can justify the use of icatibant.


Assuntos
Angioedema/induzido quimicamente , Angioedema/tratamento farmacológico , Bradicinina/análogos & derivados , Idoso , Antialérgicos/uso terapêutico , Anti-Inflamatórios não Esteroides/uso terapêutico , Bradicinina/uso terapêutico , Antagonistas de Receptor B2 da Bradicinina/uso terapêutico , Feminino , Humanos , Uso Off-Label
6.
Int J Immunopathol Pharmacol ; 27(3): 449-54, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25280038

RESUMO

Primary Snoring (PS) has been positioned at the milder end of the Sleep-Disordered Breathing severity continuum characterized by snoring and it is usually underestimated. PS is defined as snoring without apnea, frequent arousals, or gas exchange abnormalities and recent studies demonstrated that children with PS have increased blood pressure and reduced arterial distensibility. The association between adipokines and SDB has been recently investigated, though most of the studies were focused on OSAS where intermittent hypoxia characterizing the disease may lead to an inflammatory cascade and to the release of several adipokines, contributing to oxidative stress. Resistin, initially described s an adipokine increasing insulin resistance, has been recently identified as a novel important member of the cytokine family involved in the regulation of inflammation. The aim of our study was to investigate circulating resistin levels in normal weight children with PS. Sixty-five children of normal weight aged between 4 and 14 years of age were selected for habitual snoring. Children with positive polysomnography were excluded from the study. Serum resistin levels were detected in all children with PS. Thirty-three healthy non-snorer children with similar age, sex and BMI were selected as a control group. A significantly higher level of resistin was observed in patients with PS compared to the control group (4.67±1.91 ng/ml vs 3.98±1.58 ng/ml; p<0.01). Patients with inconclusive pulse oximetry showed significantly higher resistin levels than those with negative recordings recordings (5.29±1.91 ng/ml vs 4.20±1.93 ng/ml; p<0.008). Moreover, there was a significant increasing trend between sieric adipokine level and the frequency of snoring (p<0.006). Our results suggest that systemic inflammation and oxidative stress may also play a significant role in the pathophysiology of PS.


Assuntos
Resistina/sangue , Ronco/sangue , Adolescente , Criança , Pré-Escolar , Feminino , Humanos , Inflamação/complicações , Masculino , Estresse Oxidativo , Apneia Obstrutiva do Sono/sangue , Ronco/etiologia
7.
Eur J Immunol ; 31(7): 2016-25, 2001 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-11449354

RESUMO

The activation of protein tyrosine kinase(s) (PTK) is a critical event required for the development of NK cell-mediated cytotoxicity. Here we demonstrate that the adaptor protein shc undergoes tyrosine phosphorylation during the generation of antibody-dependent cellular cytotoxicity (ADCC) and natural killing. In addition, we report that, upon direct or antibody-dependent target cell interaction, shc coprecipitates with the Src homology 2 (SH2)-containing inositol phosphatase, SHIP. To gain information on the functional role of shc in NK cytotoxicity, we overexpressed wild-type or dominant negative shc constructs in the human NKL cell line. Our findings show a consistent shc-mediated down-regulation of ADCC and natural killing. Such functional effect correlates with a perturbation of the phosphoinositide (PI) metabolism by means of a shc-mediated negative regulation of inositol 1,4,5 triphosphate (IP3) generation and intracellular calcium flux upon CD16 ligation. Furthermore, our data show that dominant-negative shc-mediated perturbation of shc/SHIP interaction leads to inhibition of ligand-dependent SHIP recruitment to CD16 zeta chain. We suggest that shc plays a role of negative adaptor by modulating SHIP recruitment to activation receptors involved in the generation of NK cytotoxic function.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal , Proteínas Adaptadoras de Transporte Vesicular , Citotoxicidade Imunológica , Células Matadoras Naturais/imunologia , Proteínas/fisiologia , Citotoxicidade Celular Dependente de Anticorpos , Sinalização do Cálcio , Linhagem Celular , Células Cultivadas , Testes Imunológicos de Citotoxicidade , Humanos , Células K562 , Substâncias Macromoleculares , Fosfatidilinositol-3,4,5-Trifosfato 5-Fosfatases , Monoéster Fosfórico Hidrolases , Fosfotirosina/metabolismo , Proteínas/genética , Receptores de IgG/imunologia , Proteínas Adaptadoras da Sinalização Shc , Transdução de Sinais , Proteína 1 de Transformação que Contém Domínio 2 de Homologia de Src , Transfecção , Domínios de Homologia de src
8.
J Exp Med ; 192(1): 77-85, 2000 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-10880528

RESUMO

During the early stages of thymopoiesis, cell survival is controlled by cytokines that regulate the expression of antiapoptotic proteins such as Bcl-2. At the pre-T cell stage, a critical checkpoint for beta chain selection is monitored by the tumor suppressor p53: pre-T cells can survive and differentiate when p53 is removed genetically or when its proapoptotic function is inactivated physiologically as a consequence of signaling through the pre-T cell receptor complex. Previous work has shown that the guanine nucleotide binding protein Rho controls cell survival in T cell progenitors. Here we define the survival pathways controlled by Rho in pre-T cells and show that this GTPase is a pivotal regulator of the p53-mediated checkpoint operating at the time of beta selection: loss of Rho function results in apoptosis in pre-T cells, but this cell death is prevented by loss of p53. The prevention of cell death by loss of p53 restored numbers of early T cell progenitors but did not fully restore thymic cellularity. Further analysis revealed that loss of Rho function caused survival defects in CD4/8 double-positive thymocytes that is independent of p53 but can be prevented by ectopic expression of Bcl-2. These studies highlight that the GTPase Rho is a crucial component of survival signaling pathways in at least two different thymocyte subpopulations: Rho controls the p53 survival checkpoint in pre-T cells and is also crucial for a p53 independent survival signaling pathway in CD4/8 double positives.


Assuntos
Toxinas Botulínicas , Linfócitos T/fisiologia , Proteína Supressora de Tumor p53/metabolismo , Proteínas rho de Ligação ao GTP/metabolismo , ADP Ribose Transferases/genética , ADP Ribose Transferases/metabolismo , Animais , Apoptose , Sobrevivência Celular , Clostridium botulinum/enzimologia , Clostridium botulinum/genética , Genes bcl-2 , Genes p53 , Humanos , Camundongos , Camundongos Transgênicos , Reação em Cadeia da Polimerase , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Linfócitos T/citologia , Linfócitos T/imunologia , Timo/citologia , Timo/imunologia
9.
J Immunol ; 162(6): 3148-52, 1999 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-10092764

RESUMO

Here we investigate the activation of and a possible role for the hematopoietic Rac1 exchange factor, Vav, in the signaling mechanisms leading to NK cell-mediated cytotoxicity. Our data show that direct contact of NK cells with a panel of sensitive tumor targets leads to a rapid and transient tyrosine phosphorylation of Vav and to its association with tyrosine-phosphorylated Syk. Vav tyrosine phosphorylation is also observed following the activation of NK cells through the low-affinity Fc receptor for IgG (Fc gamma RIII). In addition, we demonstrate that both direct and Ab-mediated NK cell binding to target cells result in the activation of nucleotide exchange on endogenous Rac1. Furthermore, Vav antisense oligodeoxynucleotide treatment leads to an impairment of NK cytotoxicity, with Fc gamma RIII-mediated killing being more sensitive to the abrogation of Vav expression. These results provide new insight into the signaling pathways leading to cytotoxic effector function and define a role for Vav in the activation of NK cell-mediated killing.


Assuntos
Proteínas de Ciclo Celular , Citotoxicidade Imunológica , Proteínas de Ligação ao GTP/fisiologia , Células Matadoras Naturais/imunologia , Proteínas Proto-Oncogênicas/fisiologia , Transdução de Sinais/imunologia , Animais , Testes Imunológicos de Citotoxicidade , Proteínas de Ligação ao GTP/metabolismo , Humanos , Células K562 , Células Matadoras Naturais/metabolismo , Ativação Linfocitária , Camundongos , Fosforilação , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Proto-Oncogênicas c-vav , Receptores de IgG/fisiologia , Células Tumorais Cultivadas , Tirosina/metabolismo , Proteínas rac de Ligação ao GTP
10.
J Immunol ; 159(8): 3767-73, 1997 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-9378963

RESUMO

Fc gamma RIII (CD16) is a hetero-oligomeric receptor composed of a ligand-binding alpha subunit associated with homo- or heterodimers of the TCR zeta- and Fc epsilon RI gamma-chains. We have previously demonstrated that CD16 ligation promotes complex formation between tyrosine-phosphorylated shc and Grb2, leading to activation of ras signaling pathway in human NK cells. Here we report that CD16 engagement induces rapid shc association with the tyrosine-phosphorylated receptor complex in human NK cells. In vitro binding studies demonstrate that this interaction is mediated by the shc-SH2 domain, and immunodepletion experiments indicate that the zeta- but not the gamma-chain has the capability to mediate this association. Jurkat cell clones expressing CD16-zeta or -gamma homodimers have been used to gain more information about the mechanism of shc/CD16 association. Our data show that, while engagement of both receptors induces tyrosine phosphorylation of shc and Grb2 recruitment, shc-SH2/receptor complex association is evident only in CD16-zeta but not in CD16-gamma transfectants. Overall, our data demonstrate that the adaptor protein shc can be recruited to the activated CD16 complex by interaction with tyrosine-phosphorylated zeta-chain in a SH2-dependent manner. These results also provide further support to the notion that zeta- and gamma-chains might couple to different biochemical pathways.


Assuntos
Células Matadoras Naturais/metabolismo , Proteínas de Membrana/metabolismo , Receptores de Antígenos de Linfócitos T gama-delta/metabolismo , Receptores de Antígenos de Linfócitos T/metabolismo , Receptores de IgG/metabolismo , Tirosina/metabolismo , Domínios de Homologia de src/imunologia , Células Cultivadas , Humanos , Células Jurkat , Ligantes , Fosforilação , Ligação Proteica/imunologia , Transfecção/imunologia
11.
Immunity ; 7(1): 163-74, 1997 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-9252129

RESUMO

Mice lacking thymic function of the GTPase Rho show severe defects in fetal and adult thymopoiesis. Rho thymi are deficient in CD44+ CD25+ pro-T cells and CD44- CD25+ early pre-T cells because Rho function is required for survival but not G1/S phase cell cycle progression in these populations. The selective apoptosis defect in Rho prothymocytes can be rescued by expression of a bcl-2 transgene. A second function for Rho is seen in CD44- CD25- late pre-T cells: Rho regulates cell cycle progression but not survival of this population. These studies show that the critical processes of proliferation and survival are independently regulated during thymopoiesis and establish two different functions for Rho in the development of early thymic progenitors.


Assuntos
Proteínas de Ligação ao GTP/fisiologia , Proteínas Ativadoras de GTPase , Linfócitos T/citologia , Timo/citologia , Envelhecimento/imunologia , Envelhecimento/metabolismo , Animais , Linfócitos T CD4-Positivos/citologia , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/citologia , Linfócitos T CD8-Positivos/imunologia , Sobrevivência Celular , Citometria de Fluxo , Células-Tronco Hematopoéticas/citologia , Células-Tronco Hematopoéticas/imunologia , Camundongos , Camundongos Transgênicos , Proteínas Proto-Oncogênicas c-bcl-2/genética , Receptores de Antígenos de Linfócitos T alfa-beta/metabolismo , Receptores de Antígenos de Linfócitos T gama-delta/metabolismo , Timo/embriologia , Transgenes
12.
EMBO J ; 16(9): 2397-407, 1997 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-9171353

RESUMO

The present study employs a genetic approach to explore the role of Rho GTPases in murine thymic development. Inactivation of Rho function in the thymus was achieved by thymic targeting of a transgene encoding C3 transferase from Clostridium botulinum which selectively ADP-ribosylates Rho within its effector domain and thereby abolishes its biological function. Thymi lacking functional Rho isolated from C3 transgenic mice were strikingly smaller and showed a marked (90%) decrease in cellularity compared with their normal litter mates. We also observed a similar decrease in levels of peripheral T cells in C3 transgenic mice. Analysis of the maturation status of thymocytes indicated that differentiation of progenitor cells to mature T cells can occur in the absence of Rho function, and both positive and negative selection of T cells appear to be intact. However, transgenic mice that lack Rho function in the thymus show maturational, proliferative and cell survival defects during T-cell development that severely impair the generation of normal numbers of thymocytes and mature peripheral T cells. The present study thus identifies a role for Rho-dependent signalling pathways in thymocyte development. The data show that the function of Rho GTPases is critical for the proliferative expansion of thymocytes. This defines a selective role for the GTPase Rho in early thymic development as a critical integrator of proliferation and cell survival signals.


Assuntos
Toxinas Botulínicas , Proteínas de Ligação ao GTP/fisiologia , Proteínas Ativadoras de GTPase , Timo/crescimento & desenvolvimento , ADP Ribose Transferases/genética , ADP Ribose Transferases/metabolismo , Animais , Apoptose , Linfócitos T CD4-Positivos/citologia , Linfócitos T CD8-Positivos/citologia , Ciclo Celular , Diferenciação Celular , Clostridium botulinum/enzimologia , Feminino , Proteínas de Ligação ao GTP/antagonistas & inibidores , Antígeno H-Y/genética , Antígeno H-Y/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Receptores de Antígenos de Linfócitos T/genética , Receptores de Antígenos de Linfócitos T/metabolismo , Timo/citologia , Transgenes
13.
Eur J Immunol ; 26(12): 2807-11, 1996 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-8977272

RESUMO

We recently showed evidence of CD44-mediated enhancement of natural killer (NK) cell cytotoxic activity and induction of intracellular Ca2+ flux. In this study, we evaluated whether CD44 plays a stimulatory role in NK cell functions, such as cytokine production and activation antigen expression. Our results indicate that ligation of the CD44 receptor results in the induction of expression of the CD69 surface activation antigen as well as in the enhancement of phorbol ester-induced TNF-alpha secretion. We report also evidence for the coupling of CD44 receptor to a protein tyrosine kinase(s) pathway. CD44 engagement rapidly stimulates the tyrosine phosphorylation of several cellular substrates. Pretreatment of NK cells with the tyrosine kinase inhibitor herbimycin A resulted in marked decrease of CD44-stimulated phosphorylation, indicating that it activates tyrosine kinase(s). Furthermore, the drug also prevents CD44-mediated TNF-alpha production and CD69 expression. These findings indicate that protein tyrosine phosphorylation is an early and critical event in CD44-mediated activation of NK cell functions.


Assuntos
Receptores de Hialuronatos/efeitos dos fármacos , Receptores de Hialuronatos/metabolismo , Células Matadoras Naturais/efeitos dos fármacos , Ativação Linfocitária/efeitos dos fármacos , Proteínas Tirosina Quinases/farmacologia , Antígenos CD/biossíntese , Antígenos de Diferenciação de Linfócitos T/biossíntese , Células Cultivadas , Reagentes de Ligações Cruzadas/farmacologia , Humanos , Células Matadoras Naturais/metabolismo , Lectinas Tipo C , Fosforilação , Transdução de Sinais/efeitos dos fármacos , Acetato de Tetradecanoilforbol/farmacologia , Fator de Necrose Tumoral alfa/biossíntese
14.
J Exp Med ; 183(1): 179-86, 1996 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-8551221

RESUMO

The Src homology (SH) 2/SH3 domain-containing protein Grb2 and the oncoprotein Shc have been implicated in a highly conserved mechanism that regulates p21ras activation. We investigated the involvement of these adaptor proteins in the signaling pathway induced by CD16 or interleukin (IL) 2R triggering in human natural killer (NK) cells. Both p46 and p52 forms of Shc were rapidly and transiently tyrosine phosphorylated upon CD16 or IL-2 stimulation with different kinetics. Shc immunoprecipitates from lysates of CD16- or IL-2-stimulated NK cells contained Grb2 and an unidentified 145-kD tyrosine phosphoprotein. Grb2 immunoprecipitates from anti-CD16-stimulated NK cells contained not only Shc, but also a 36-kD tyrosine phosphoprotein (p36). The interaction between Grb2 and Shc or p36 occurred via the Grb2SH2 domain as indicated by in vitro binding assays using a bacteriologically synthesized glutathione S-transferase-Grb2SH2 fusion protein. We also present evidence that p21ras is activated by CD16 and IL-2R cross-linking. Accumulation of guanosine triphosphate-bound Ras was detected within 1 minute and occurred with kinetics similar to inductive protein tyrosine phosphorylation and Grb2 association of Shc and p36 adaptor proteins.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal , Proteínas Adaptadoras de Transporte Vesicular , Células Matadoras Naturais/fisiologia , Proteínas/metabolismo , Proteínas Proto-Oncogênicas p21(ras)/metabolismo , Receptores de IgG/metabolismo , Proteína Adaptadora GRB2 , Humanos , Immunoblotting , Interleucina-2/farmacologia , Células Matadoras Naturais/efeitos dos fármacos , Fosfoproteínas/metabolismo , Fosforilação , Testes de Precipitina , Ligação Proteica , Receptores de Interleucina-2/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Proteínas Adaptadoras da Sinalização Shc , Transdução de Sinais/fisiologia , Proteína 1 de Transformação que Contém Domínio 2 de Homologia de Src , Tirosina/metabolismo , Domínios de Homologia de src
15.
Nat Immun ; 15(2-3): 147-53, 1996.
Artigo em Inglês | MEDLINE | ID: mdl-9162264

RESUMO

NK cells are endowed with a wide array of adhesion molecules which mediate their interaction with endothelium and extracellular matrix components. We have shown that cross-linking of beta 1 integrins and CD44 on human NK cells induces a signal transduction pathway involving both tyrosine kinase activation and the modulation of intracellular calcium levels. Our studies have also demonstrated the ability of beta 1 integrins and CD44 to upregulate both the spontaneous and the CD16-triggered cytotoxic activity of human NK cells. Although the molecular mechanisms responsible for this costimulatory activity have not been defined yet, our studies indicate that the simultaneous cross-linking of beta 1 integrins and CD44 results in a synergistic effect on CD16-mediated enhancement of intracellular free calcium concentration, and suggest that this may be relevant for the cooperation observed.


Assuntos
Comunicação Celular/imunologia , Citotoxicidade Imunológica , Matriz Extracelular/imunologia , Líquido Intracelular/imunologia , Células Matadoras Naturais/imunologia , Transdução de Sinais/imunologia , Humanos
16.
J Immunol ; 153(10): 4399-407, 1994 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-7525702

RESUMO

CD44, a major hyaluronate receptor, is involved in a variety of lymphocyte functions including lympho-hemopoiesis, adhesion to high endothelial venules or the extracellular matrix, and T cell activation. Here we investigated the ability of CD44 to affect the cytotoxic functions of human NK cells. Ligation of CD44 by selected mAb (J173 and F10442) resulted in a rapid, dose-response-dependent enhancement of NK cytotoxic activity against a panel of tumor target cells that varied in their sensitivity to NK killing. Neither enhanced killing against NK-resistant target cells nor CD44 mAb-mediated redirected lysis was not observed. CD44 cross-linking also was found to up-regulate CD16-mediated lysis. In an attempt to investigate the early biochemical events that occur after CD44 ligation, we found that optimal cross-linking conditions induce a rapid increase of intracellular free calcium levels, which is abrogated by extracellular Ca2+ chelation. Moreover, enhanced and more sustained Ca2+ rise resulted from CD16 and CD44 coengagement. In contrast, no inositol 1,4,5-trisphosphate generation was found after optimal CD44 cross-linking. These results suggest that although CD44 is not capable of delivering a lytic signal in human NK cells, it coactivates spontaneous or CD16-mediated NK cytotoxicity. The variation in intracellular free calcium may be one of the signals that account for the costimulation of the lytic activity.


Assuntos
Citotoxicidade Celular Dependente de Anticorpos/imunologia , Proteínas de Transporte/imunologia , Células Matadoras Naturais/imunologia , Receptores de Superfície Celular/imunologia , Receptores de Retorno de Linfócitos/imunologia , Anticorpos Monoclonais/imunologia , Cálcio/metabolismo , Testes Imunológicos de Citotoxicidade , Citometria de Fluxo , Humanos , Receptores de Hialuronatos , Inositol 1,4,5-Trifosfato/biossíntese , Ativação Linfocitária , Transdução de Sinais/imunologia , Células Tumorais Cultivadas
17.
J Immunol ; 153(1): 21-31, 1994 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-7515923

RESUMO

Lymphohematopoiesis, cell matrix adhesion, homing of leukocytes, T cell activation, and tumor metastasis are mediated through the CD44 family of cell surface receptors. We have recently shown that anti-CD44 mAb trigger protein tyrosine kinase-dependent activation of T cell effector functions. Here, we show that hyaluronate (HA), a CD44 ligand, in conjunction with CD3/TCR-mediated stimuli, is costimulatory for human peripheral blood T cell proliferation, for IL-2 production by Th clones, and for release of trypsin-like esterase by cytolytic T cell clones. A human T cell line, HUT-78, was found to bind HA and on HA coating it was used as a target for cytolytic T cell clones. After anti-CD3 stimulation, CD3+/CD8+ clones acquire the ability of lysing HA-coated HUT-78 cells more efficiently than the same HA-uncoated targets. Resting peripheral blood T cells and T cell clones do not adhere to HA-coated plates. However, 24-h anti-CD3 mAb stimulation gives them the transient ability to bind HA. HA adhesion of activated T cells and T cell clones, as well as that of T cell lines, is blocked by one anti-CD44 mAb (J-173). Two other anti-CD44 mAbs induce a 10-fold increase in HA adhesiveness of anti-CD3-stimulated peripheral blood T cells. This impressive HA adhesiveness is also readily blocked by J-173 anti-CD44 mAb. These data indicate that 1) HA is costimulatory for human T cell effector functions in conjunction with CD3/TCR-mediated stimuli, 2) the capacity to bind HA is acquired by resting T cells and T cell clones after anti-CD3 stimulation, and 3) HA binding occurs via specific interaction with CD44 molecules expressed on activated T cells.


Assuntos
Proteínas de Transporte/metabolismo , Ácido Hialurônico/farmacologia , Receptores de Superfície Celular/metabolismo , Receptores de Retorno de Linfócitos/metabolismo , Subpopulações de Linfócitos T/imunologia , Linfócitos T CD4-Positivos/imunologia , Adesão Celular/efeitos dos fármacos , Humanos , Receptores de Hialuronatos , Ácido Hialurônico/metabolismo , Interleucina-2/biossíntese , Ativação Linfocitária , Subpopulações de Linfócitos T/citologia , Linfócitos T Citotóxicos/efeitos dos fármacos , Linfócitos T Auxiliares-Indutores/efeitos dos fármacos
18.
Bone Marrow Transplant ; 11(5): 357-61, 1993 May.
Artigo em Inglês | MEDLINE | ID: mdl-8504269

RESUMO

Rejection of T-depleted BMTs is predominantly mediated by alloreactive host T cells. A low but significant number of radiochemoresistant clonable T cells can be detected following a conventional cytoreductive protocol given prior to T-depleted BMT. Elimination of these cells increases the engraftment rate. We found no clonable T cells at the end of the conditioning regimen in 100 ml of peripheral blood from 47 patients who received an HLA-identical T-depleted BMT. None rejected the graft and none displayed mixed chimerism. In addition, although no clonable T cells were detected in nine patients who received a mismatched BMT, two rejected their graft. However, in three mismatched patients, who for clinical reasons received a modified pre-BMT schedule, the presence of host clonable T cells was associated with immunological rejection. These findings suggest that the detection of clonable T cells should prove a valuable indicator for optimising immunosuppression prior to T-depleted BMT.


Assuntos
Purging da Medula Óssea , Transplante de Medula Óssea/imunologia , Rejeição de Enxerto/imunologia , Linfócitos T/imunologia , Adolescente , Adulto , Criança , Testes Imunológicos de Citotoxicidade , Feminino , Rejeição de Enxerto/sangue , Rejeição de Enxerto/etiologia , Antígenos HLA/imunologia , Teste de Histocompatibilidade , Humanos , Masculino , Pessoa de Meia-Idade
19.
J Immunol ; 150(10): 4225-35, 1993 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-8097750

RESUMO

mAb against the lymphocyte homing receptor CD44/Hermes up-regulate the proliferation of human T PBL induced by anti-CD3 or anti-CD2 mAb. Moreover, certain anti-CD44 mAb can activate human resting T cells and mouse cytotoxic T cells in the absence of anti-CD3 or anti-CD2 mAb. Here, we show that anti-CD44 mAb trigger proliferation of human CD3+/CD4+ T cell clones in a fashion similar to that observed with mAb to CD3. Such an effect is IL-2-dependent, as shown by IL-2 production induced by anti-CD44 mAb and by complete inhibition of cell proliferation in the presence of anti-IL-2 antibodies or cyclosporin A. Moreover, anti-CD44 mAb trigger human cytolytic T cell clones to lyse Fc gamma-R+ P815 cells in the absence of additional stimuli. The magnitude of the cytolytic response induced by anti-CD44 mAb is comparable to that observed in the presence of anti-CD3 mAb for both CD4+ and CD8+ TCR-alpha/beta+ clones, and for V delta 1 or V delta 2 TCR-gamma/delta+ clones. By contrast, in CD3-/CD16+ NK cell clones, no cytolytic responses to anti-CD44 mAb could be observed. Granule trypsin-like esterase enzyme (granzyme) release by cytolytic T cell clones is induced by plastic-immobilized anti-CD44 mAb. Anti-CD44 mAb-triggered proliferation ([3H]thymidine incorporation) and cytotoxicity are blocked by the protein tyrosine kinase inhibitor, genestein. In addition, ligation of the CD44 molecule induces tyrosine phosphorylation of proteins identical, by molecular mass, to those phosphorylated after anti-CD3 mAb stimulation. Notably, anti-CD44 mAb does not induce tyrosine phosphorylation of a 21-kDa protein (the phosphorylated zeta-chain of the TCR molecular complex) typically observed upon anti-CD3 mAb stimulation. In conclusion, this study shows that the ligated CD44 molecule provides the necessary stimuli for a variety of T cell-mediated functions triggered via protein tyrosine kinase-dependent signal transduction pathways at least in part similar to those that follow stimulation of the CD3/TCR complex.


Assuntos
Receptores de Retorno de Linfócitos/imunologia , Subpopulações de Linfócitos T/imunologia , Linfócitos T CD4-Positivos/citologia , Linfócitos T CD4-Positivos/imunologia , Células Clonais , Citotoxicidade Imunológica/efeitos dos fármacos , Genisteína , Humanos , Técnicas Imunológicas , Técnicas In Vitro , Isoflavonas/farmacologia , Ativação Linfocitária , Proteínas Tirosina Quinases/antagonistas & inibidores , Proteínas Tirosina Quinases/farmacologia , Transdução de Sinais
20.
Cytotechnology ; 11 Suppl 1: S100-2, 1993.
Artigo em Inglês | MEDLINE | ID: mdl-7763733

RESUMO

We show that antibodies to the CD44 molecule trigger proliferation of human CD3+/CD4+ T-cell clones. Such effect is IL2-dependent, as shown by IL2 production induced by anti-CD44 mAb and by inhibition of cell proliferation in the presence of anti-IL2 antibodies or cyclosporin A (CsA). Moreover, anti-CD44 mAb triggered human cytolytic CD4+ and CD8+ TCR alpha/beta+ clones, and V delta 1 or V delta 2 TCR Y/delta+ clones to lyse Fc-gamma-R+ P815 cells and to release granule trypsin-like esterase enzymes. Anti-CD44 mAb-triggered proliferation and cytotoxicity were blocked by the PTK-inhibitor, genestein. In addition, ligation of the CD44 molecule induced tyrosine phosphorylation of proteins identical, by molecular weight, to those phosphorylated following anti-CD3 mAb-stimulation. Notably, anti-CD44 mAb does not induce tyrosine phosphorylation of a 21 kD protein (the phosphorylated zeta chain of the TcR molecular complex) typically observed upon anti-CD3 mAb stimulation.


Assuntos
Receptores de Retorno de Linfócitos/imunologia , Linfócitos T Citotóxicos/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Anticorpos Monoclonais , Biotecnologia , Divisão Celular , Células Clonais/citologia , Células Clonais/imunologia , Humanos , Ativação Linfocitária , Proteínas Tirosina Quinases/imunologia , Receptores de Retorno de Linfócitos/antagonistas & inibidores , Transdução de Sinais/imunologia
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