Superoxide dismutases in idiopathic pulmonary fibrosis.
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Adult↗
Biomedical subjects
A Facchini
Publications and source records attributed to A Facchini.
Endothelin-1 in idiopathic pulmonary fibrosis.
AIMS--To evaluate whether endothelin-1 is involved in the pathology of idiopathic pulmonary fibrosis (IPF). METHODS--Plasma endothelin-1 concentrations were evaluated in 37 patients with IPF and 27 normal controls by radioimmunoassay. In addition, expression of endothelin-1 in lung tissue was evaluated in biopsy specimens obtained from four patients with IPF. Three biopsy specimens of normal lung were used as controls. Endothelin-1 immunoreactivity was detected using immunohistochemistry. RESULTS--Elevated endothelin-1 plasma concentrations were found in patients with IPF compared with controls and a positive correlation was found with duration of disease. No significant difference was observed between treated and untreated patients with IPF. Increased endothelin-1 immunoreactivity was found in lungs of three of four patients with IPF. Endothelin-1 positive consisted mainly of small vessel endothelial cells. Some scattered macrophages were also positive. CONCLUSIONS--Elevated plasma concentrations and expression of endothelin-1 in lung tissue are suggestive of increased production of endothelin-1 in at least a proportion of patients with IPF. Consequently, endothelin-1 activity could play a role in the fibrogenic process of the disease.
Adult↗
Distribution and lytic activity of NK cell subsets in the elderly.
Old subjects present an increased number of NK cells associated with a decreased lytic activity of isolated and cloned CD16 cells. Recently, two new surface molecules of 58 kDa, identified by the monoclonal antibodies GL183 and EB6, have been described. The presence of these molecules, which can be coexpressed on CD16+ cells allows the recognition of the NK cell subsets whose cytolytic activity is restricted to different allospecificities. This study investigated a group of old subjects to determine whether a particular distribution or a different lytic activity of NK subsets, defined by MoAbs GL183 and EB6, is involved in the altered cytolytic activity found during ageing. Further, we investigated whether the ageing process might be responsible for a restriction of the NK cell repertoire involved in the recognition of allogenic cells. We found that old and young subjects have a similar proportion of double positive and double negative GL183/EB6 cells, while in the old group single positive subsets were increased. The lytic activity of sorted NK subsets isolated from old and young subjects was similar, although double positive and double negative cells from the old presented a lower cytotoxic activity. The addition of IFN-beta or rIL-2 to the culture medium restored the lytic activity to the level found in young subjects. These data show that the decreased NK lytic activity found in the old subjects is shared out among the different NK subsets and normal aged subjects do not lose the NK repertoire found in the young.
Aged↗
Standardization of a micro-cytotoxicity assay for human natural killer cell lytic activity.
Cytotoxicity assays are widely used to evaluate the functional activity of NK and T cells against tumour target cells and the release of radioactive sodium chromate from labelled target cells is still the most commonly used marker of target lysis in culture supernatants. We describe here the standardization of a micro-cytotoxicity test in which the number of cytolytic effector and tumour target cells have been decreased by a factor of 10. The release obtained by 500 tumour target cells was compared with the release obtained by 5000 target cells in the standard cytotoxicity assay for target:effector cell ratios from 1:1 to 1:100. Both gamma and beta emissions of the 51Cr isotope were evaluated to determine the assay release. The results obtained by the micro-cytotoxicity assay (500 target cells) were comparable to those of the standard assay (5000 target cells) and 51Cr release evaluation using the gamma counter was the most sensitive method of determining lytic activity using 500 tumour target cells. beta counter evaluation using solid phase scintillation was found to be a reproducible alternative method, even if the lytic curves cannot be compared with those obtained using the traditional method.
Chromium↗
Intracellular Cu/Zn superoxide dismutase levels in T and non-T cells from normal aged subjects.
The decreased immune response associated with aging may, in part, reflect intrinsic age-related biochemical alteration in lymphocytes from older subjects. The 'reactive oxygen species hypothesis' of aging postulates that these molecules are involved in the modifications leading to cellular senescence. Superoxide dismutase (SOD), and in particular the Cu/Zn-dependent intracellular form, plays a critical role in the defense against these species, but it is controversial whether this function declines in lymphocytes in old age. We utilized two different methods to evaluate Cu/Zn SOD levels in T and non-T cells (CD3+, CD3-, CD4+, CD8+, CD16+) from young and old individuals: a specific and sensitive enzyme immunoassay performed on extracts of sorted cells, and a flow cytometry double fluorescence technique with monoclonal antibodies against Cu/Zn SOD and the different lymphocyte subsets. The Cu/Zn SOD cell content was assayed both in basal conditions and after peripheral blood lymphocyte stimulation with Concanavalin A, anti-CD3 monoclonal antibody and phorbol myristate acetate. In basal conditions, and considering the various subsets, no differences were found between young and old individuals, although data analysis revealed high and low responders in both groups. Taking all the subjects together, higher levels of this enzyme were found in CD3+ T lymphocytes, in particular in the CD4+ cells. After peripheral blood lymphocyte stimulation, Cu/Zn SOD concentration was higher than levels in unstimulated cells, both in young and old individuals, and particularly using Concanavalin A with respect to anti-CD3 and phorbol myristate acetate. In conclusion, the synthesis of Cu/Zn SOD does not seem to be affected by aging in proliferating cells. The highest levels of Cu/Zn SOD present in CD4+ cells, both from young and old individuals, may prevent the oxidant stress of these elements which play a major role in the inflammation sites.
Adult↗
Synergism between gamma interferon and doxorubicin in a human MDR colon adenocarcinoma cell line.
Recent interest in cancer therapy derives from the ability of interferons to synergistically increase the activity of chemotherapeutic agents. To understand the biological basis of this synergism we evaluated the effects of human recombinant IFN-gamma on the expression of the mdr1 gene and on the cellular growth of a human colon adenocarcinoma cell line (LoVo) and its MDR subline (LoVo/Dx) after coincubation with doxorubicin. Treatment with IFN-gamma showed unchanged levels of MDR1-glycoprotein, no perturbation on cell cycle distribution and a significant reduction of colony formation in both lines (P < 0.05) starting from 100 U/ml. A synergistic effect was observed in the LoVo/Dx cell line when doxorubicin was added after exposure to 0.1-10 U/ml of IFN-gamma. Our data indicate that the effects of IFN-gamma, independent from action on cell proliferation and from modulation of p-glycoprotein expression, are a cause of the synergistic activity between this lymphokine and conventional chemotherapeutic agents such as doxorubicin.
ATP Binding Cassette Transporter, Subfamily B, Mem↗
Elevated soluble intercellular adhesion molecule 1 in the serum of patients with polymyalgia rheumatica: influence of steroid treatment.
OBJECTIVE: To determine the levels of serum soluble intercellular adhesion molecule 1 (sICAM-1) in patients with polymyalgia rheumatica (PMR) and/or giant cell arteritis (GCA) prior and during steroid therapy. METHODS: Sixteen patients with PMR and/or GCA had complete clinical and laboratory investigations at diagnosis before treatment and 9 patients were followed during 6 months of steroid therapy. Serum sICAM-1 levels were measured by ELISA: RESULTS: Serum ICAM-1 levels were higher in the patient group compared to a control population. No significant correlation was found between serum sICAM-1 levels and other indices of disease activity. After steroid administration, a dramatic fall in serum sICAM-1 levels was observed at the end of the first week and persisted until the end of the study period, paralleling clinical and laboratory disease remission. CONCLUSION: Serum sICAM-1 levels paralleled disease activity in PMR and/or GCA. Steroid treatment induced a rapid normalization of the serum sICAM-1 levels.
Aged↗
Ischemia-reperfusion syndrome: an alternative experimental model.
OBJECTIVE: To design an alternative experimental model of ischemia-reperfusion syndrome. Our model mimics the clinical pattern of the syndrome and also assesses the efficacy of therapeutical protocols. EXPERIMENTAL DESIGN: Ischemia was induced under general anaesthesia in the posterior limbs of 10 sheep by occluding the aorta and vena cava by means of two-way balloon catheters. Ischemia was stopped after 4 hours and blood and histologic parameters determined in the first three hours of revascularization. The animals were divided into three groups: a group of 3 sheep in which a sham operation was performed; a control group (5) to assess the efficacy of induced ischemia; the third group (5) to determine the effect of antioxidant and membrane protective drugs to assess the reliability of the model to study the ischemia-reperfusion syndrome. RESULTS: At the end of ischemia, skin temperature was decreased (p < 0.04) both in control and treated groups, pH decreased significantly soon after reperfusion in the control group (p < 0.04). Reperfusion in control sheep, compared with treated animals, was followed by a significant increase in CPK blood levels (p < 0.009), related to marked muscle damage, in particular after reperfusion. Tissue damage detected at TEM was less severe in treated animals. CONCLUSIONS: This model is an effective experimental strategy and a means of assessing preventive treatment.
Animals↗
Nutritional status affects the sensitivity to anoxia/reoxygenation in hepatocytes.
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Adenosine Triphosphate↗
Flow cytometry and in vitro tritiated thymidine labeling in normal rectal mucosa of patients at high risk of colorectal cancer.
OBJECTIVES: To compare two different methods to evaluate rectal epithelial cell proliferation as a biomarker of risk of developing colon cancer. METHODS: Samples of normal rectal mucosa from 26 patients at increased risk for colorectal cancer (22 patients with adenoma, three with adenocarcinoma of the large bowel, and one with longstanding ulcerative colitis) were examined by means of in vitro labeling with tritiated thymidine and flow cytometry. RESULTS: We found a significant correlation between thymidine-labeling index and the percentage of cells in S-phase, measured by flow cytometry both in formalin-fixed, paraffin-embedded specimens and in frozen specimens (respectively, r = 0.7647, p < 0.001, and r = 0.4503, p < 0.01). However, using flow cytometry, the percentage of cells in S-phase was significantly higher than the thymidine-labeling index in both fixed-embedded and frozen specimens (p < 0.01). Proliferative parameters were not higher in patients with colon carcinoma, and were not related to the degree of dysplasia, the number of adenomas, or familial occurrence of colorectal cancer. Two specimens taken from normal rectal mucosa of two patients with adenomas showed aneuploidy. No aneuploidy was found in normal rectal specimens of patients with adenocarcinoma. CONCLUSIONS: These results show that the calculation of cells in S-phase with in vitro tritiated thymidine labeling or by flow cytometry produces different results. However, the significant correlation between corresponding parameters obtained with these techniques support the use of either method as "intermediate biomarkers" of colorectal cancer risk and prognosis.
Adenocarcinoma↗
Elevated interleukin-8 serum concentrations in beta-thalassemia and graft-versus-host disease.
Neutrophil chemotactic and functional defects occur in beta-thalassemia and in patients after bone marrow transplantation (BMT). Interleukin-8 (IL-8) is a novel chemotactic and activating peptide for neutrophils and can be detected in the circulation. IL-8 serum concentrations were evaluated in 30 beta-thalassemic patients before and after BMT. Serial samples from 16 patients were also studied. Fourteen sera from healthy children, 43 patients with chronic viral hepatitis, 16 patients on chronic transfusion treatment for various hematologic disorders, and 28 healthy adults were studied as controls. IL-8 was evaluated by an enzyme-linked immunosorbent assay. Patients with beta-thalassemia had higher IL-8 concentrations than did normal controls, patients with liver disease, and patients on chronic transfusion. beta-Thalassemic patients with severe liver siderosis and fibrosis had the highest IL-8 concentrations. After BMT in patients with successful engraftment, IL-8 concentrations decreased significantly. In contrast, in patients with acute graft-versus-host disease (GVHD), IL-8 concentrations were not statistically different from the concentrations found before BMT and were higher than in patients with no complications and patients with graft rejection. IL-8 may play a part in the immune dysregulation that occurs in beta-thalassemia and may be involved in the immune mechanisms leading to GVHD.
Adolescent↗
Antibodies to topoisomerase II in idiopathic pulmonary fibrosis.
Antibodies to DNA topoisomerase II (anti-topoisomerase II) were detected by ELISA in the sera of 18 out of 41 (44%) patients with idiopathic pulmonary fibrosis (IPF). Follow-up sera were also obtained from 19 of the patients. DNA topoisomerase II binding remained constantly high or low in the majority of follow-up sera, but 2 out of the 8 positive cases became negative while 3 out of the 11 negative cases became positive during follow-up. No association was found between occurrence of anti-topoisomerase II antibodies and any indices of disease severity. Furthermore, individual patient follow-up did not show any correlation between changes in topoisomerase II binding and deterioration or improvement of clinical status. In conclusion our study shows that although anti-topoisomerase II are detectable in a large fraction (approximately 50%) of IPF patients and are useful for diagnostic purposes, they do not provide a measure of clinical activity.
Adult↗
Decreased adhesion to endothelial cells and matrix proteins of H-2Kb gene transfected tumour cells.
Transfection of murine metastatic B78H1 cells (derived from B16 melanoma) with a syngeneic H-2Kb gene was used to study the effect of Major Histocompatibility Complex (MHC) gene products on tumour cell adhesion to endothelial cells and matrix proteins and the involvement in the metastatic process. H-2Kb-expressing transfectants showed a reduced adhesion to endothelial surfaces of different origin (four murine endotheliomas and human umbilical vein endothelial cells) when compared to parental B78H1 cells and to controls transfected with pSV2neo alone. On the average a 50-70% reduction in adhesion to endothelial cells was observed among H-2Kb transfectants. H-2Kb transfectants had a reduced expression of the alpha 4 integrin subunit, moreover the adhesion of Neo-transfected clones to endothelial cells was reduced to the levels of H-2Kb transfectants by antibodies directed against the beta 1 subunit and the endothelial VCAM-1 molecule, thus suggesting an impairment of the VLA-4/VCAM-1 interaction in H-2Kb transfectants. Adhesion to extracellular matrix components was also strongly decreased: in general the adhesion of H-2Kb cells showed a 50-75% inhibition with respect to Neo or parental controls. The highest difference was observed in adhesion to vitronectin and laminin, the lowest in adhesion to fibronectin. Reduction in adhesive properties of H-2Kb-expressing transfectants could be involved in the reduced metastatic ability, evaluated by means of intravenous injection of cells: H-2Kb transfectants yielded less than ten lung colonies, while all controls produced more than 100. Our data indicate that expression of a single class I MHC gene can significantly alter the metastatic phenotype of MHC-negative tumour cells and this could be related to a general alteration of tumour cell adhesive interactions.
Animals↗
Elevated serum superoxide dismutase levels correlate with disease severity and neutrophil degranulation in idiopathic pulmonary fibrosis.
1. Tissue damage in idiopathic pulmonary fibrosis is due in part to oxidant-antioxidant imbalance. 2. We evaluated the serum levels of the antioxidant enzyme Cu/Zn superoxide dismutase (EC 1.15.1.1) in 25 patients with idiopathic pulmonary fibrosis, 34 patients with sarcoidosis and 40 healthy control subjects by an enzyme immunometric assay. 3. We found that patients with idiopathic pulmonary fibrosis have higher serum Cu/Zn superoxide dismutase levels than control subjects and patients with sarcoidosis. In addition, serum Cu/Zn superoxide dismutase levels correlate with disease severity indexes in patients with idiopathic pulmonary fibrosis. 4. The increase in serum Cu/Zn superoxide dismutase level in idiopathic pulmonary fibrosis could depend on degranulation of activated neutrophils or release from damaged cells. To elucidate the contribution of neutrophil degranulation we determined the polymorphonuclear cell elastase level in the same specimens. We found a strong correlation between serum Cu/Zn superoxide dismutase and polymorphonuclear cell elastase activities, and, in patients with idiopathic pulmonary fibrosis, we observed higher levels of polymorphonuclear cell elastase than in control subjects and patients with sarcoidosis, which correlated positively with disease severity indexes. 5. Cu/Zn superoxide dismutase can catalyse the dismutation of O2 into H2O2 and generate OH.. These oxygen radicals are probably the major factors responsible for tissue damage (in particular, alveolar and endothelial cells) and fibrosis in experimental lung injury.(ABSTRACT TRUNCATED AT 250 WORDS)
Adult↗
Rectal cell proliferation as an intermediate biomarker of risk of colorectal cancer.
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Biomarkers↗
In vitro immunotoxicity of +/- 2'-deoxy-3'-thiacytidine, a new anti-HIV agent.
The present study compares the in vitro effect of (+/-)-2'-deoxy-3'-thiacytidine (BCH 189) a new synthetic anti-HIV-1 dideoxynucleoside, with 3'-azido-3'-deoxythymidine (AZT) on the immune function of lymphocytes from 10 normal and 12 HIV-1+ patients (CDC II and III). The effect of different doses of BCH 189 and AZT was analysed in vitro on: (i) T cell proliferation after stimulation with concanavalin A (Con A) or anti-CD3 MoAb; (ii) B cell proliferation and immunoglobulin production after stimulation with pokeweed mitogen (PWM); (iii) cytokine production (IL-2, IL-6, GM-CSF, tumour necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma) from lymphocytes stimulated with anti-CD3 MoAb or phytohaemagglutinin (PHA). BCH 189 inhibited the proliferation of B and T lymphocytes from normal and HIV+ subjects less than AZT; even if lymphocytes from HIV+ (CDC III) subjects produced higher levels of IL-6 and TNF-alpha, neither BCH 189 nor AZT molecule interfered with cytokine release. Immunoglobulin production from B lymphocytes was inhibited only by a high concentration (50 microM) of BCH 189 or AZT. These results show that BCH 189 affects lymphocyte proliferation in vitro less than AZT, and support its use in clinical trials in HIV-infected patients.
Antiviral Agents↗
Detection of circulating autoantibodies to poly(ADP-ribose)polymerase in autoimmune diseases.
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Autoantibodies↗
Natural killer function in flow cytometry. III. Surface marker determination of K562-conjugated lymphocytes by dual laser flow cytometry.
The recognition of effector cell populations that are able to actively from conjugates with target cells is of major importance in studies of lymphocyte cytotoxicity. A number of methodologies have been described to identify the conjugates and count them, but there have been few studies of the binding capability of the different subsets of effector cells involved in the conjugation phenomenon. Here we describe a methodology that permits the study of two surface markers on lymphocytes conjugated to K562 target cells. In particular, the expression of low density CD8 (CD8dim) has been studied on both CD3+ and CD16+ lymphocytes bound to K562 target cells. Previously described methodologies, either optical microscopy or flow cytometry, were not able to identify the effector population by mAb double staining, especially in the case of antigens expressed at low density. The flow cytometric methodology described here permits the measurement of the binding activity of small lymphocyte subsets such as the CD3+ 8dim+ population. However, the method could be used to study the binding activity of any effector population defined by mAb double staining.
Antigens, Differentiation↗