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J Carracedo

Publications and source records attributed to J Carracedo.

At least 19 recordsLinked to original sources
PubMed2006

[Acute rejection of kidney transplant is associated with induction of replicative senescence in CD8+ T lymphocytes].

Acute renal rejection repeatedly activates immunocompromised CD8 + T cells. Maintained activation of CD8 + T cells can induce a process of replicative senescence. In the present study, we will evaluate in CD8 lymphocytes from patients undergoing acute renal rejection characteristics of replicative senescence such as: a) low expression of CD28 molecule; b) telomere shortening and c) increase production of proinflammatory cytokines. The study was carried out in CD8 + T cells from 14 patients transplanted without clinical evidences of acute renal rejection, 14 patients kidney transplanted with clinical and anatomopathological evidences of acute renal rejection, 8 healthy controls. The results shown that in peripheral blood and renal biopsy of patients with acute renal rejection there is a significant increment of the population of T cells CD28-CD8+, with short telomere length, as compared with healthy controls and patients without acute renal rejection. The presence of senescent cells was associated with high levels of IL-10 and IFN-Y in plasma and urine. In conclusion our study suggest that the CD8 + T cells of patients with acute renal rejection suffer a process of replicative senescence.

Adult↗
PubMed2001

Antithrombin III prevents early pulmonary dysfunction after lung transplantation in the dog.

BACKGROUND: Ischemia-reperfusion injury with the resulting inflammatory response is a devastating complication of lung transplantation; much of the tissue damage could be diminished by control of the inflammatory response. Recent studies have show that antithrombin III (AT III) has an anti-inflammatory effect in addition to its established role in the regulation of blood coagulation. Thus, we hypothesized that the administration of AT III might help to prevent ischemia-reperfusion injury after lung transplantation. METHODS AND RESULTS: The study was performed in a dog model of orthotopic lung transplantation. Dogs were randomly assigned to receive either vehicle (controls) or AT III. We observed that in control dogs, during the 180-minute period after lung transplantation, the arterial O(2) partial pressure decreased and both the alveolar-arterial O(2) difference and the pulmonary vascular resistance increased. By contrast, these parameters remained unchanged in the group of dogs receiving AT III. Dogs with transplants receiving AT III did not show an increase in cell adhesion molecules, and histological examination revealed almost an absence of inflammatory response. The administration of AT III produced a marked increase in serum prostacyclin (PGI(2)) levels, whereas in control dogs, the PGI(2) levels did not change. The beneficial effect of AT III was not observed when dogs received indomethacin to prevent the stimulation of PGI(2) release by AT III. CONCLUSIONS: Our results demonstrate that AT III prevents ischemia-reperfusion injury in a dog model of lung transplantation and that this effect is conditioned by an increase in PGI(2) production.

6-Ketoprostaglandin F1 alpha↗
PubMed2001

TH2 lymphocytes from atopic patients treated with immunotherapy undergo rapid apoptosis after culture with specific allergens.

BACKGROUND: In atopic patients treatment with specific immunotherapy (SIT) induced a shift in the balance of T-cell immune response away from a T(H)2-type (producing mostly IL-4) in favor of a T(H)1-type T-lymphocyte response (with the preferential production of IFN-gamma). However, the mechanisms through which SIT acts are less clear. We have recently shown that allergens may induce an activation-induced cell death process in lymphocytes from SIT-treated atopic patients. OBJECTIVE: This study aimed to determine whether allergen-induced apoptosis can occur in a specific subset of cells. METHODS: The study was performed in lymphocytes from normal subjects and atopic patients, some of whom were treated with SIT. Cells were cultured in the presence of gramineous pollen (Lolium perenne) allergenic extracts. Cell phenotype and intracellular cytokine expression were measured by means of fluorescent mAbs. Apoptosis was measured by using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling. Fluorescence was analyzed in a FACScan flow cytometer, and the data were evaluated with Consort 30 software. RESULTS: Our results showed that allergens induce apoptosis of lymphocytes in SIT-treated atopic patients. Apoptosis occurs mainly in T(H)2 lymphocytes with the IL-4+/CD4+ phenotype and subsequently increases the percentage of IFN-gamma(+) cells in the culture. CONCLUSION: These results suggest that the shift from T(H)2 to T(H)1 induced by SIT in atopic patients may be mediated, at least in part, by the induction of an activation-induced cell death process in allergen-responder T(H)2 cells.

Allergens↗
PubMed2001

Role of adhesion molecules in mononuclear cell apoptosis induced by cuprophan hemodialysis membranes.

BACKGROUND/AIM: Hemodialysis with Cuprophan (CU) membranes induces mononuclear cell activation, leading to increased expression of adhesion molecules, formation of cell aggregates, and apoptosis. It is likely that structure(s) of the CU membrane interact with mononuclear cell surface molecules which transduce biochemical signals to the cell. Interactions between adhesion molecules and extracellular matrix have been implicated in cell activation, proliferation, and/or apoptosis. In the present work, we study whether adhesion molecules may be involved in CU-induced mononuclear cell aggregation and/or apoptosis. METHODS: The present study was performed using THP-1 cells, a human monocytic cell line, cultured in the presence of the CU membrane. CD11b and CD54 expression was studied with fluorescent monoclonal antibodies. Cell aggregation was quantified using a phase-contrast microscope. Apoptosis was evaluated by either light microscopy or annexin V labeling. RESULTS: The results show that incubation of CU membranes with the proteins CD11b, CD18, and CD54 or the blockade of these cell surface molecules with specific monoclonal antibodies inhibited the CU-induced aggregation and apoptosis in a dose-dependent manner. CONCLUSION: These results suggest that CU membranes interact selectively with these specific proteins to induce cell activation which ultimately results in apoptosis.

Apoptosis↗
PubMed1999

Allergens induce apoptosis in lymphocytes from atopic patients.

Repeated stimulation of immune cells may induce an "activation-induced cell death" (AICD) program. Allergy is characterized by the cyclic activation of allergen-reactive immune cells. To study the effects of allergen stimulation in cell proliferation and apoptosis in atopic subjects, peripheral blood mononuclear cells (PBL) from 40 atopic patients with positive reactivity to the allergens Olea Europaea (OE) and Lollium Perenne (LP) (20 without immunotherapy and 20 with specific immunotherapy) and 10 normal subjects were cultured with the allergens OE and LP. PBL from atopic patients proliferate more vigorously than cells from normal subjects after culture in vitro with both allergens, although PBL from atopic subjects without immunotherapy proliferate more than PBL from atopic subjects with immunotherapy. The study of cell proliferation shows that in atopic patients PBL mainly exhibit the CD4/CD45RO phenotype. This preferential proliferation is more evident in PBL from atopic patients treated without immunotherapy. Cell culture with specific allergens induces apoptosis in PBL from atopic patients. The percentage of apoptosis increased when atopic patients had been previously treated with immunotherapy. In addition to the observed increase in cell proliferation, apoptosis mainly occurs in the CD45RO cells that support the involvement of these cells in allergy. Furthermore, results obtained in cells from immunized patients suggest that an AICD process may partly at least explain the mechanism of action of allergen immunotherapy.

Allergens↗
PubMed1999

NK phenotypic markers and IL2 response in NK cells from elderly people.

Immunosenescence is a process that primarily affects the T cell compartment of the immune system, although age-associated immunological alterations have also been demonstrated in the NK cell phenotype and function. A significant expansion in the number of NK cells is found in aging. The NK cytotoxic capacity of total peripheral blood lymphocytes is also well preserved, not only in healthy elderly people but also in centenarians. However, NK cell killing of K562 is impaired when considered in a per-cell basis, and this defect is associated with defective signal transduction after activation more than a diminished conjugate formation or killing capacity. We have studied the phenotype of NK cells in elderly donors fulfilling the Senieur criteria. We have also studied the capacity of these cells to be activated by IL2 when different NK cell functions, other than cytotoxicity, are considered. Our results confirm the increased percentage of NK cells in the elderly due to the expansion of the CD56dim subset that also show an altered pattern of activation markers, whereas no differences were found in the CD56bright subset. The response of NK cells to IL2 was found to be impaired when proliferation, expression of CD69, and Ca2+ mobilization were considered, whereas TNF-alpha production was not significantly affected. These results suggest that human NK cells do not escape the aging process, although senescence have a differential effect on distinct NK cell biological functions, ranging from severe to negligible impairment, depending on the parameters considered.

Adult↗
PubMed1998

Serum levels of intercellular adhesion molecule 1 (ICAM-1) in patients with colorectal cancer: inhibitory effect on cytotoxicity.

A positive correlation between the level of ICAM-1 in serum and the stage of neoplastic processes has been demonstrated. We studied ICAM-1 serum concentration in 27 colorectal cancer patients and investigated the effect of this molecule on cellular aggregation and toxicity. ICAM-1 serum concentration in the group of patients was significantly higher (P < 0.01) than in normal controls and was related to tumour stage. Patient sera inhibited both the formation of cellular aggregates and the percentage of specific lysis, the effect being lost when the serum was depleted of ICAM-1. These results suggest that the release of soluble ICAM-1 may represent a mechanism of tumour escape.

Aged↗
PubMed1998

Nonbiocompatible hemodialysis membranes induce apoptosis in mononuclear cells: the role of G-proteins.

Mononuclear cells are activated during hemodialysis. In this study, we provide evidence that in vitro culture of mononuclear cells with Cuprophan, a nonbiocompatible hemodialysis membrane, increases the levels of protein phosphorylation in these cells as well as the expression of surface activation molecules. By contrast, culturing of mononuclear cells with AN69, a more biocompatible membrane, did not increase protein phosphorylation levels or expression of surface activation molecules in these cells. In addition, Cuprophan, but not the AN69 membrane, increased the percentage of mononuclear cell death by apoptosis. Inhibition of G-protein-mediated signal transduction decreased the apoptosis of cells cultured with the Cuprophan membrane. The GTP-binding protein involved in Cuprophan-induced apoptosis was sensitive to the ADP-ribosylating pertussis toxin (PTX). The inhibition of GTP-binding protein decreased apoptosis in the early stage of the activation-induced apoptosis, suggesting that G-proteins are implicated in the transmission of apoptosis-inducing signal(s) but do not interfere with the effector signals that mediate the late stages of apoptotic catabolism. Finally, PTX was capable of inhibiting apoptosis without affecting expression of activation molecules; thus, the inhibition of apoptosis by Cuprophan was not due to quenching of the stimulation signals, because monocytes were still able to be activated by Cuprophan despite the action of PTX. The results obtained in this study suggest that cell activation and apoptosis may be mediated by separate intracellular signals.

Acrylic Resins↗
PubMed1997

Mycobacterium tuberculosis induces apoptosis in gamma/delta T lymphocytes from patients with advanced clinical forms of active tuberculosis.

Antigens from inactivated Mycobacterium tuberculosis H37Ra induce activation in a subpopulation of gamma/delta (gamma/delta) T lymphocytes in a manner that resembles that of superantigens from alpha/beta T cells. After culture in vitro with H37Ra proteins, gamma/delta T lymphocytes from patients with advanced clinical forms of active tuberculosis (ACF-TBC) display cytotoxic activity against homotypic target cells exposed to H37Ra. Cytotoxicity by gamma/delta T lymphocytes from ACF-TBC patients occurs in a range similar to that observed in healthy subjects. Following activation, H37Ra-stimulated gamma/delta T lymphocytes from healthy subjects did proliferate in the presence of exogenous recombinant human interleukin 2. However, under the same conditions, gamma/delta T lymphocytes from ACF-TBC patients not only did not proliferate but died by apoptosis. These results suggest that in gamma/delta T lymphocytes from patients with ACF-TBC, antigens from M. tuberculosis may induce cell activation that leads to apoptotic cell death.

Antigens, Bacterial↗
PubMed1997

C1-esterase inhibitor prevents early pulmonary dysfunction after lung transplantation in the dog.

The success of lung transplantation to a large extent depends on effective protection of the graft from ischemic injury after reperfusion. Although mechanisms have not been clarified, the pathologic findings of ischemic injury after reperfusion are similar to adult respiratory distress syndrome, a condition in which the blood coagulation contact system is activated. This study evaluates the effect of C1-esterase inhibitor (C1-INH), the main inhibitor of the blood coagulation contact system, on short-term lung function in a dog model of orthotopic lung transplantation. Twelve lung transplantations were performed after 24 h of ischemic time. Dogs were randomly assigned to receive either vehicle (Control) or C1-INH. After the lung transplantation in the control group, Pao2 decreased by 84% and both the AaPO2 and the Qs/Qt% increased (340 and 530%, respectively, p < 0.01); these parameters remained unchanged in the C1-INH group. The hypoxemia observed in control animals was associated with decreased blood coagulation contact factors, complement consumption, increased expression of adhesion glycoproteins in leukocytes, and extensive intraalveolar and interstitial neutrophil infiltration. In contrast, C1-INH administration prevented hypoxemia, the decrease in blood coagulation contact factors, the activation of the complement system, the increase in expression of leukocyte adhesion molecules, and inflammatory cell infiltrate. This study has demonstrated that in a dog model of lung transplantation, the administration of C1-INH prevents early pulmonary dysfunction, and it suggests that activation of blood coagulation contact system and complement are important mechanisms causing ischemic injury after reperfusion.

Animals↗
PubMed1996

CD69-induced monocyte apoptosis involves multiple nonredundant signaling pathways.

Simultaneous stimulation of human monocytes/macrophages or THP1 cells with LPS and an antibody specific for the activation marker CD69 induces apoptosis. Here we demonstrate the involvement of multiple independent signals that are necessary for apoptosis induction. Thus, inhibitors of phospholipase A2 and lipoxygenase prevent apoptosis induction. Similarly, the ADP-ribosylating G-protein-reactive pertussis toxin (PTX) but not a mutant toxin lacking the ADP-ribosylating moiety (mPTX) prevents apoptosis induction. Furthermore, inhibition of NO generation abrogates completely the induction of apoptosis by LPS/CD69 ligation. These three pathways can be dissociated from each other in the sense that interventions on the arachidonic acid metabolism or G proteins do not inhibit the generation of NO and that exogenous NO cannot reverse the inhibition of cell death by inhibitors of phospholipase A2 or PTX. In addition, both PTX and mPTX affect arachidonic acid mobilization only partially, indicating that the apoptosis-inhibitory effect of PTX (which is not shared by mPTX) cannot be explained by its effect on phospholipase A2 activation. Both LPS and anti-CD69 are sufficient on their own to activate cells, as determined by TNF production, NO generation, or arachidonic acid metabolism, but neither LPS nor anti-CD69 can induce apoptosis on their own. Thus, apoptosis induction in this system involves at least three independent signal transduction systems--(i) arachidonic acid metabolism, (ii) NO, and (iii) PTX-sensitive events--each of which is necessary but insufficient to induce monocyte/macrophage apoptosis. These findings underline the complex control of activation-induced apoptosis in cells of the myelomonocytic lineage.

Adult↗
PubMed1996

1 alpha, 25-dihydroxyvitamin D3 (calcitriol) induces apoptosis in stimulated T cells through an IL-2 dependent mechanism.

The steroid hormone 1 alpha,25-dihydroxyvitamin D3 (calcitriol) has been shown to inhibit PHA-induced T-cell proliferation. In the present work we show that calcitriol caused a concentration-dependent decrease in the viability of PHA stimulated but not unstimulated T lymphocytes. Under microscopy, nonviable cells exhibit classic features of programmed cell death or apoptosis. In addition, the same concentration of calcitriol induces a blockage in the G1 phase of T cell cycle and decreased secretion of IL-2, which was correlated with the extent of apoptosis. Thus, addition of exogenous IL-2 reverted both the cell cycle block and the apoptotic cell death induced by calcitriol. This data indicates that calcitriol induced apoptosis in PHA-stimulated T cell, and suggests that this action is mediated through suppression of IL-2 secretion.

Animals↗
PubMed1996

Inhibition of apoptotic cell death in B-CLL by interferon gamma correlates with clinical stage.

Apoptosis was evaluated in B cells from 41 patients with B-CLL and 20 healthy aged-matched controls. B cells were cultured with and without gamma-IFN and other cytokines; apoptosis was quantified at regular intervals throughout a 5-day culture period. According to Rai's criteria, 17 patients were classified as good risk, 16 as intermediate and eight as high risk. In vitro, purified B cells from B-CLL patients were evaluated for apoptosis. Maximal apoptosis (44.12%) was observed at day 5 in cells from patients with poor prognosis. The addition of gamma-IFN to the culture media prevented apoptosis in a dose-dependent manner. Maximal inhibition of apoptosis was achieved with 100 IU/ml of gamma-IFN. The degree of inhibition of apoptosis by gamma-IFN was greater in cells from the high-risk group patients than in those from the intermediate and good prognosis group (P < 0.0001). The expression of gamma-IFN receptors in B-CLL cells was evaluated using a MnAb against the extracellular domain of gamma-IFN receptor. After 4 days in culture with gamma-IFN, only cells from the intermediate- and high-risk groups showed an increase in the density of gamma-IFN receptors (P < 0.001). gamma-IFN was not detected in the sera of our study patients. However gamma-IFN was detectable in the media from both normal B cells and B-CLL cells in culture; there was no difference in the amount of gamma-IFN released by cells from the three groups of patients studied. Our results show that in vivo gamma-IFN inhibits apoptosis of B cells from B-CLL patients. The inhibitory effect of gamma-IFN on apoptosis correlates directly with the severity of the disease and this is likely explained by a marked upregulation of gamma-IFN receptors in cells from patients in the high-risk group.

Aged↗
PubMed1995

Pertussis toxin-sensitive GTP-binding proteins regulate activation-induced apoptotic cell death of human natural killer cells.

Apoptosis of natural killer (NK) cells can be induced by non-specific physical damage (UV irradiation, heat shock) or by simultaneous ligation of the CD16 and the interleukin-2 receptor (IL-2R) molecules, but not with either anti-CD16 or IL-2 alone. Whereas blockade of GTP-binding protein (G protein)-mediated signal transduction using ADP-ribosylating bacterial toxins or the GTPase-resistant GTP analog guanosine 5'-0-(3-thiotriphosphate (GTP gamma S) does not affect non-specific induction of NK cell apoptosis, such interventions do inhibit induction of apoptosis by anti-CD16/IL-2. The G proteins involved in the regulation of activation-induced NK apoptosis are sensitive to pertussis toxin (PTX) and to the non-specific GTP analog GTP gamma S but not to cholera toxin, Pseudomonas exotoxin A or diphtheria toxin. A pertussis toxin mutant that lacks ADP-ribosylating activity, but conserves the membrane translocating and T cell-mitogenic effects of the native molecule, fails to inhibit NK apoptosis. To exert their apoptosis-inhibitory effect, PTX and GTP gamma S must be employed before cells are activated. Later addition has no effect, suggesting the implication of G proteins in the transmission of apoptosis-inducing signals, but not in the effector stage of apoptosis. Pre-incubation with PTX or GTP gamma S does not affect the activation of NK cells by CD16 cross-linking, IL-2 stimulation- or both, as assessed by the induction of CD69 expression, protein tyrosine phosphorylation and calcium mobilization. Moreover, neither PTX nor GTP gamma S compromise the effector function of NK cells or the susceptibility of target cells to NK-mediated lysis. These data suggest apoptosis as a novel mechanism by which NK responses may be controlled in vivo, as well as an experimental and therapeutical strategy to counteract endogenous down-regulation of NK responses.

Apoptosis↗
PubMed1995

Cell aggregation and apoptosis induced by hemodialysis membranes.

During hemodialysis, circulating mononuclear cells can be stimulated to different degrees, depending on membrane biocompatibility. Cell activation usually leads to aggregation and proliferation. It may also result in apoptosis if cells are subjected to abnormal activation. This may be the case of cells exposed to bioincompatible hemodialysis membranes. The study presented here evaluates the effects of two hemodialysis membranes, with different degrees of biocompatibility, (Cuprophan (CU; Lundia IC 5N; GAMBRO, Sweden) and polyacrylonitrile (AN69; Biospal 3000S, Hospal, France)) on aggregation and apoptosis of circulating human mononuclear cells and the human mononuclear cell line (THP-1). The results showed that 2-h incubation with CU, a bioincompatible membrane, produces cell aggregation of both peripheral mononuclear cells and THP-1 cells (35% and 54%, respectively). After 48 h of incubation with a CU membrane, apoptotic death was observed in 32% of THP-1 cells and in 45% of normal peripheral mononuclear cells. Neither cell aggregation nor apoptosis was observed after incubation with the AN69 membrane. CU membrane-induced apoptosis was inhibited by Staurosporrin (Sigma, St. Louis, MO) a protein kinase C (PKC)-inhibitor, suggesting that cell apoptosis induced by the CU membrane is mediated by a PKC-dependent cell activation. Furthermore, cell prestimulation with phorbol 12-myristate 13-acetate, an activator of PKC, results in a increase in the percentage of THP-1 cell death by apoptosis after CU exposure (53%). Our study indicates that CU membranes induce mononuclear cell activation, leading to cell apoptosis.

Acrylic Resins↗