Interleukin-6, interferon-gamma, and phospholipid levels in the alveolar lining fluid of human lungs. Profiles in coal worker's pneumoconiosis and idiopathic pulmonary fibrosis.

Cytokines are widely involved in physiologic as well as immunoinflammatory and fibrosing processes of the lung. The aim of this work was to study, by bronchoalveolar lavage, two groups of human interstitial lung diseases (ILD) with fibrosing propensity (ie, idiopathic pulmonary fibrosis [IPF], n = 10; and coal worker's pneumoconiosis [CWP], n = 15). Patients were compared with nonsmoker control subjects (n = 20). Cellularity, proteins, and phospholipids were determined in the alveolar fluids. In addition, two cytokines (interleukin-6 [IL-6] and interferon-gamma [IFN-gamma]), which are presumed to possess respective antifibrotic and profibrotic activities, were measured in the respiratory tract. Compared with control subjects, IPF and simple CWP showed alveolar hypercellularity (p < 0.05) and relative lymphocytosis (p < 0.05). Both exhibited increased alveolar permeability (ie, increased albumin/urea ratio, p < 0.05), with enhanced IL-6 and decreased IFN-gamma in the alveolar spaces (p < 0.05). On the other hand, IPF displayed an associated polymorphonuclear alveolitis, enhanced alveolar epithelial lining fluid (AELF) volume and low surfactant phospholipid levels (p < 0.05 vs control), whereas simple CWP shared an exclusive lymphocytosis, normal AELF volume, and a surfactant lipid overflow (p < 0.05 vs control). Relationships among all of these parameters were found only between alveolar cellularity, neutrophils and IL-6 levels in the AELF of IPF (respectively, r = 0.85, p = 0.0009, and r = 0.89, p = 0.0006). In summary, common alterations of cellular and cytokine turnover were observed in IPF and simple CWP and may reflect activity of the antifibrotic fight in these diseased lungs. Surfactant phospholipid levels are likely to represent a specific disturbance among IPF and CWP, but no clear relationship with respect to the other parameters could be established for explaining the difference in time course outcome.

Bone marrow transplantation as treatment for X-linked immunodeficiency with hyper-IgM.

We report a 10-year-old boy with a severe form of immunodeficiency with hyper-IgM who underwent successful bone marrow transplantation with his HLA-matched sister as donor. Busulfan (20 mg/kg) and cyclophosphamide (200 mg/kg) were used as conditioning. The post-transplant course was uneventful. He is alive 25 months later with full hematological and immunological reconstitution.

Effects of indomethacin and diclofenac on some functions of polymorphonuclear neutrophils.

The effects in-vitro of two non-steroidal anti-inflammatory drugs, diclofenac and indomethacin, have been studied on human polymorphonuclear functions. Adhesivity and chemotaxis were decreased by the two drugs, but in a manner that varied with the attractant used. As shown by the nitro blue tetrazolium slide test, all cells remained active but the production of oxygen metabolites by opsonized zymosan was decreased in the presence of diclofenac.

Influence of heparin on the chemotactic activity of human thrombin.

Chemoattractant properties of human thrombin have been studied, by polymorphonuclear leucocyte migration under agarose gel, in the presence of various sulphated macromolecules such as standard heparins, low molecular weight heparins, CY216, K2165, PK10169 and pentosane polysulphate. These compounds did not attract polymorphonuclear leucocytes within the range of concentrations used, whilst thrombin alone is a cytotaxin for these cells. Addition of heparins to thrombin led to an increase in the chemoattractant activity of this enzyme for at least one of the doses studied. Augmentation of the chemoattractant activity of thrombin by heparins was shown at concentrations equivalent to those found in-vivo after administration of therapeutic doses of heparin. Pentosane polysulphate, at the studied concentrations, did not lead to a significant rise in the chemoattractant activity of thrombin.

[Effect of cholesterol on the cellular deformability and osmotic fragility of erythrocytes]. / Influence du cholestérol sur la déformabilité cellularire et la fragilité osmotique des hématies.

The effect of cholesterol hemisuccinate incorporation on the rheological properties of the erythrocyte membrane was assessed by measuring deformability (elongation index) and osmotic fragility ("baseline hemolysis"). These two experimental methods have different response sensitivities according to the incubation protocols (12 h 25 degrees C/12 h 25 degrees C then 12 h 37 degrees C) ant to the amount of cholesterol incorporated in the membrane (C/Pt from 0.18 to 0.83). The deformability and osmotic fragility variations observed are inversely related with the cholesterol concentrations incorporated. The experimental differences observed in the deformability and osmotic fragility are discussed in view of the type of membrane deformation and of the signals collected by the two systems.

[Acute leukemia secondary to the treatment of multiple sclerosis with chlorambucil]. / Leucémie aiguë secondaire au traitement de la sclérose en plaque par le chlorambucil.

A new case is reported here of acute myeloid leukemia after chlorambucil therapy for multiple sclerosis. This is the sixth case seen at our institution during the period 1978 to 1984. The leukemogenic potential of the immunosuppressive cytotoxic drugs in patients with multiple sclerosis is emphasized.

[Evaluation of the Coulter Counter S+IV LMG: estimation of the leukocyte formula. Comparison with microscopic methods]. / Evaluation du Coulter Counter S+IV LMG: estimation de la formule leucocytaire. Comparaison avec des méthodes microscopiques.

The evaluation of a differential count instrument of the Coulter S+line, the S+IV LMG Coulter-Counter, in particular for the lymphocyte, monocyte and granulocyte count was carried out. This differential analysis was compared with more classical methods of the manual (microscope) or automatic type (image analyser, Hematrack 360 type, Geometric Data). The repeatability is very good and the correlations excellent. The instrument can even detect the presence of low percentages of abnormal cells. Placement of reagents and samples at room temperature is important and the samples can be kept at room temperature for at least 9 hours without disturbing the count. In addition, energetic shaking of the samples does not change the count for the three types of cells. Due to the alarms given by the instrument as regards the cell distributions and all of the parameters of the blood cell count, no pathology should be ignored. In their laboratory, the authors note that about 25% of the smears are further examined by microscope.

Analysis of blood CD4+ and CD8+ T-lymphocyte subsets with monoclonal antibodies. Comparison of the Genetic Systems CD4/CD antigen typing kit with conventional indirect immunofluorescence microscopy and the automated Technicon-H1 Enzyme Immunoassay (EIA).

The Genetic Systems Technique (GS: direct immunofluorescence microscopy with ethidium bromide counterstaining of nuclei) was tested for quantitative analysis of T-lymphocyte subsets in human peripheral blood. The monoclonal antibodies anti-CD4 and anti-CD8 were used for detection of T-helper and T-suppressor cells respectively and the results compared to those obtained by conventional indirect immunofluorescence microscopy and the Technicon Enzyme Immunoassay (EIA) with automated reading. The GS technique provided results correlating well with both indirect immunofluorescence and EIA techniques. Moreover, this method has two advantages: it is less time-consuming than the indirect immunofluorescence microscopy and necessitates less expensive and more commonly available equipment than the automated EIA technique.

[Changes in hemogram parameters in infections]. / Variations des paramètres de l'hémogramme au cours des infections.

PURPOSE: Leukocytosis is considered as an argument for infection. We have compared leukocytosis with the other data available with the white blood cell count. METHODS: White blood cell counts obtained from 187 patients (age: 18 to 81 years), admitted to an emergency room for abdominal pain, were analysed using an automate (Bayer Technicon H2, Dublin, Ireland), and compared with matched healthy subjects. The patients were classified into two groups: diseases of the biliary or of the urinary tract, and subsequently in two subgroups: infectious diseases (angiocholitis, cholecystis, pyelonephritis) or non-infectious diseases (hepatic and nephritic colic). RESULTS: Leukocytes and neutrophils were significantly increased, and eosinophils significantly decreased in all subjects by comparison with controls. These abnormalities were more important in infected patients. Lymphocytes were significantly decreased in infectious disease. For predicting infection, sensitivity and specificity of leukocytosis (> 1,000/mm3) were respectively 66% and 56%, while that of eosinopenia (< 100/mm3) were respectively 91% and 38%, and that of lymphopenia (< 1,200/mm3) respectively 58% and 73%. The probability of infection was less than 3% when neutrophils were less than 7,000/mm3, and eosinophils and lymphocytes respectively more than 100/mm3 and 1,200/mm3. CONCLUSION: This study shows that leukocytosis, eosinopenia or lymphopenia are poor indicators of infection, when considered alone or in combination. However, eosinopenia and lymphopenia appear as better criteria of infection than leukocytosis. A detailed analysis of the white blood cell count allows the exclusion of infection with an acceptable risk of error.

[Pancytopenia resolved by the treatment of hyperthyroidism]. / Pancytopénie résolutive par le traitement d'une hyperthyroïdie.

Hyperthyroidism can be associated with various haematological disorders related to several mechanisms. These disorders might be related to the reduced life-span of whole blood components and/or to an autoimmune mechanism. Only one case of pancytopenia has yet been reported. The observation of 3 new personal cases (1 toxic adenoma and 2 Graves' disease) led us to review the pathogeny of haematological disorders found in hyperthyroidism. Only one patient had antineutrophil autoantibodies. Direct and indirect Coomb's test, and Dixon's test were negative. In all patients, bone marrow aspiration was unable to demonstrate pernicious anaemia or myelodysplastic syndrome. Two patients presented cytological signs of macrophage activation with eosinophilia. These cytological features were compatible with an immuno-allergy mechanism. All haematological disorders disappeared when patients became euthyroid. In all cases, the haematological abnormalities were quite mild and might have gone unnoticed. Thus, it can be suggested that the frequency of pancytopenia in hyperthyroidism is underestimated.

[Determination by flow cytometry of reference values of erythrocyte parameters in aged subjects]. / Détermination par cytométrie de flux des valeurs de référence de la population érythrocytaire chez le sujet âgé.

OBJECTIVES: Normal values for hemoglobin and hematocrit have been established for healthy young adults, but it is unknown whether age-related decline in hematological values occurs in the elderly. We compared normal values of hematological parameters in healthy adults and healthy persons aged 65 years and over using flow cytometry. SUBJECTS AND METHODS: Erythrocyte and reticulocyte parameters were determined in 585 subjects (294 men and 206 women aged 26 to 64 years, and 37 men and 48 women aged 65 years and over), living independently and without medication, symptom and disease, using Technicon H1 (Bayer) and R2000 (Roche) analyzers. Serum erythropoietin levels, red cell folate and creatinine clearance were determined in subjects aged 65 years and over. RESULTS: Irrespective of subject age, hemoglobin (Hb), hematocrit (Hct), red blood cell count (RBC), mean cell volume (MCV), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) were higher in males than in females, but sex-related differences tended to decrease with aging. In males, there was a trend toward lower Hb and Hct with aging, due to an increase in RBC, MCV and MCH. In females, Hb and Hct (p < 0.05) increased with aging, due to a trend toward higher values of RBC, MCV and MCH. Red blood cell distribution width and hemoglobin distribution width were not modified by aging. Despite these age-related changes in erythrocyte parameters, values for anemia defined in adults appear appropriate for geriatric subjects (Hb < 130 g/l in males and Hb < 120 g/l in females). Reticulocyte count and creatinine clearance decreased in men and women aged 65 years and over (p < 0.05), whereas erythropoietin levels and red cell folate were not influenced by aging. CONCLUSION: Normal erythrocyte values defined in adults are appropriate for evaluating hematological test results in older individuals. The decrease in reticulocyte count observed in men and women aged 65 years and over does not appear to be linked to a decrease in erythropoietin production with aging.

Electrophoretic mobility of Sézary syndrome lymphoid cells.

The variations in electrophoretic mobilities found in different lymphocyte populations present in circulating blood of three patients with Sézary syndrome have been studied. Evidence is given for variations in electrophoretic mobilities being a function of the cytological type of the pathological cells found. The existence of a significant increase of "null" cells is shown, which seem to correspond to Sézary cells.

Reticulocytes: reference limits.

Healthy reference ranges for the total peripheral reticulocyte count and its three subpopulations, LFR (low fluorescence ratio), MFR (middle fluorescence ratio) and HFR (high fluorescence ratio) were established on 1219 healthy subjects. Samples were taken from subjects of both sexes with ages ranging from 4 to over 60 years. The observed ranges were found to be different between males and females for the group over 20 years old, while there was no difference between sexes found at the age between 4 and 19 years.

Reticulocytes: biological variations and reference limits.

The reticulocyte count is a good index of erythropoietic activity. Automated methods, mainly flow cytometry, have made the counting easier, more accurate and reproducible. The data presented describe the counting of reticulocytes in 1219 apparently healthy subjects of both sexes who came for a periodic health examination. Their ages varied from 4 to over 60 years. The reticulocytes were counted with a flow cytometer (Sysmex R-3000) with Auramine-O as fluorochrome. There was no statistical difference between boys and girls aged 4-19 years. However, the reticulocyte count was significantly higher in men than in women over 20 years of age. The reticulocyte count was affected neither by the menstrual cycle in girls at puberty, nor by oral contraceptives and the menopause in women. No effect of moderate smoking was observed in either sex. Reference limits of the reticulocyte counts (percentage and absolute value) according to age and sex and the three types of reticulocytes are provided.

Leukocyte, lymphocyte and platelet response to dynamic exercise. Duration or intensity effect?

The influence of work intensity and duration on the white blood cell (WBC), lymphocyte (L) and platelet (P) count response to exercise was studied in 16 trained subjects (22 +/- 5.4 years, means +/- SD). They performed three cyclo-ergospirometric protocols: A) 10 min at 150 W followed by a progressive test (30 W/3 min) till exhaustion; B) constant maximal work (VO2max); C) a 45 min Square-Wave Endurance Exercise Test (SWEET), (n = 5). Arterial blood samples were taken: at rest, submaximal and maximal exercise in A; maximal exercise in B; 15th, 30th and 45th min in the SWEET. Lactate, [H+], PaCO2, PaO2, [Hct], Hb, cortisol, ACTH, total platelet volume (TPV), total blood red cell (RBC), WBC, L and P were measured. At 150 W, WBC, L, P, and TPV increased. VO2max did not differ between A and B, but a difference was found in total exercise time (A = 25 +/- 3 min; B = 7 +/- 2 min, p less than 0.001). In A, at VO2max, the increase was very small for Hct, [Hb], and RBC (10%), in contrast with large changes for WBC (+93%), L (+137%), P (+32%), TPV (+35%), [H+] (+39%), lactate (+715%), and ACTH (+95%). At VO2max there were no differences in these variables between A and B. During the SWEET: WBC, L, P, TPV and ACTH increased at the 15th min as much as in VO2max, but no difference was observed between the 15th, 30th and 45th min, except for ACTH which continued to rise; the lactate increase during the SWEET was about half (+341%) the value observed at VO2max, and [H+] did not vary with respect to values at rest.(ABSTRACT TRUNCATED AT 250 WORDS)

Refractory anemia with excess of blasts: a multivariate analysis of prognostic factors in 91 patients and a simplified scoring system for predicting survival.

Prognosis in myelodysplastic syndromes is extremely variable. The prognostic value of the FAB classification has been demonstrated in many studies. However, within the same FAB subtype, some patients may experience prolonged survival, whereas others die in a few weeks. This prognostic heterogeneity makes the therapy decision difficult. In an attempt to identify significant prognostic factors for survival in refractory anemia with excess of blasts (RAEB), clinical and hematological characteristics were analyzed in 91 patients. Multivariate regression analysis showed that bone marrow total blast cells percentages, sex and hemoglobin level were the characteristics significantly associated with survival. A scoring index based upon these three characteristics may be proposed and had a great prognostic value (p < 0.00001). It allows us to separate patients into three groups with low, intermediate and high score with a median survival of 239, 133 and 45 days for each group respectively. This scoring index may be useful in the design of therapy and analysis of future clinical trials. However, its predictive value needs to be confirmed in other series.

De novo methylation of tumour suppressor genes CDKN2A and CDKN2B is a rare finding in B-cell chronic lymphocytic leukaemia.

De novo methylation of the 5'CpG island has been recently reported as an alternative mechanism of inactivation for the tumour suppressor genes CDKN2A and CDKN2B. We examined CDKN2A methylation status at diagnosis in 42 B-cell chronic lymphocytic leukaemia (CLL) patients, in 19 cases the CDKN2B methylation status was also analysed. No homozygous CDKN2A/2B deletion was detected, but four patients (9%) displayed an aberrant CDKN2A methylation status and only one had hypermethylated CDKN2B. De novo methylation was associated with silencing of gene expression. These results confirm that CDKN2A/2B inactivation by deletion is a rare event in CLL and suggest that aberrant methylation could be an alternative way of inactivation very rarely involved in the development of some CLL.