Hematological recovery and peripheral blood progenitor cell mobilization after induction chemotherapy and GM-CSF plus G-CSF in breast cancer.

In order to determine the effect of GM-CSF plus G-CSF in combination in breast cancer patients receiving an effective induction regimen, we compared hematological recovery and peripheral blood progenitor cell (PBPC) mobilization according to colony-stimulating factor (CSF) support. Forty-three breast cancer patients were treated by TNCF (THP-doxorubicin, vinorelbine, cyclophosphamide, fluorouracil, D1 to D4) with CSF support: 11 patients received GM-CSF (D5 to D14); 16 patients G-CSF (D5 to D14) and 16 patients GM-CSF (D5-D14) plus G-CSF (D10-D14). Between two subsequent cycles, progenitor cells were assessed daily, from D13 to D17. The WBC count was similar for patients receiving G-CSF alone or GM-CSF plus G-CSF, but significantly greater than that of patients receiving GM-CSF alone (P<0.001). The GM-CSF plus G-CSF combination led to better PBPC mobilization, with significantly different kinetics (P<0.001) and optimal mean values of CFU-GM, CD34+ cells and cells in cycle, at D15 compared to those obtained with G-CSF or GM-CSF alone. The significantly greater PBPC mobilization obtained with a CSF combination by D15 could be of value for PBPC collection and therapeutic reinjection after high-dose chemotherapies.

Acquired angioedema with C1 inhibitor deficiency: is the distinction between type I and type II still relevant?

BACKGROUND: Acquired angioedemas are divided into type I associated with lymphoproliferation and type II caused by anti-C1-inhibitor antibodies. Recent reports have suggested that this distinction is not so clear-cut, mainly because of the presence of antibodies against the C1 inhibitor in some cases belonging to the type I group. We report herein 2 additional cases of acquired angioedema with anti-C1-inhibitor antibody. MATERIAL AND METHODS: One man and 1 woman had had acquired angioedema for several years. In the man, a monoclonal component had been detected several years before the present study. In the second patient, a monoclonal component was detected during the study. The following data were studied on successive blood samples collected during angioedema manifestations: complement component levels, functional activity of the classical pathway, functional and antigenic C1 inhibitor doses, ELISA test to detect autoantibodies to C1 inhibitor and Western blot analysis of the C1 inhibitor. RESULTS: In both patients, CH50 and C4 activities were decreased, and an autoantibody to C1 inhibitor was detected. In 1 case, the antibody appeared after the monoclonal component; in the second case, it appeared before and belonged to a different immunoglobulin class. CONCLUSION: Our data suggest that the distinction between type I and type II acquired angioedema is no longer valid because of overlapping in some cases.

Atopy parameters in asthmatic infants.

We evaluated various atopy parameters in 44 asthmatic infants aged 1-3 years: nonspecific parameters (total IgE and eosinophilia), and specific parameters relative to 21 allergens (specific IgE and skin tests), together with tests of leukotriene release by blood leukocytes (cellular allergen stimulation test; CAST) in the presence of a mixture of 21 allergens. Thus far 17 infants have displayed no sign of atopy, but 27 met at least one criterion. Eight met nonspecific criteria, and the others single or multiple criteria. Of the 21 allergens, 20 gave rise to at least one sensitization in this population. The specific IgE was more frequently positive than the skin tests. Dissociations between the two types of specific tests were practically systematic. Different phenotypes based on the chosen parameters were individualized, demonstrating heterogeneity in the expression of atopy in these young infants. The polyvalent CAST was positive only when other criteria of atopy were also positive (specific IgE and/or skin tests), and the range of intensity of the responses obtained supports reactivity rather than sensitization.

Mobilization of peripheral blood progenitor cells after induction chemotherapy (THP-doxorubicin-vinorelbine-cyclophosphamide-fluorouracil) and granulocyte colony-stimulating factor in breast cancer.

In order to evaluate the mobilization of peripheral blood progenitor cells (PBPC) after an effective induction regimen in breast cancer, we performed a study on 15 breast cancer patients. Between January 1995 and June 1996, these patients received TNCF (THP-doxorubicin. vinorelbine, cyclophosphamide, fluorouracil for four days, every 21 days) with G-CSF support (5 microg/kg for 10 days after chemotherapy) to reduce aplasia. This regimen is known to result in a complete pathological response in 30% of patients. Between two cycles of TNCF treatment, hematological recovery was observed. Progenitor cells (CFU-GM and CD34+ cells) and mononuclear cells in DNA synthesis (MCDS) counts were performed daily, between the 12th and 17th post-chemotherapy days (81 samples). The results showed a similarity for hematological recovery and PBPC mobilization kinetics depending on the number of treatment cycles. The three methods used for PBPC evaluation were well correlated (P < 0.01) with an optimal mean PBPC recruitment by the last day of G-CSF administration: respectively, 11 520 (1729-26539) CFU-GM/ml of blood, 249 (14-1160) CD34+ cells/microl of blood and 211 (21-554) MCDS/microl of blood. These results suggested that a daily injection of G-CSF after one or two TNCF cycles will produce an effective PBPC mobilization in comparison with currently used regimens.

[Mixed cryoglobulins and autoimmunity in hepatitis C]. / Cryoglobulines mixtes et auto-immunité au cours de l'hépatite C.

We explored and studied cryoglobulins and autoantibodies in 149 patients with hepatitis C. 49% of untreated patients have cryoglobuliemia (36% polyclonal cryoglobulins) and 76% of ineffectively treated patients since more than 6 months (50% polyclonal cryoglobulins). Other observed cryoglobulins are half of type II and half oligoclonal. In 22 cryoglobulinemic patients before treatment, a good clinical response to IFN is induced only among patients for which cryoglobulins are negative at the end of treatment. Cryoglobulins could be indicators of viremia. The prevalence of autoantibodies is low and without clinical significance (they are more frequently detected in cryoglobulinemic patients, except anti-thyroid autoantibodies). Nevertheless, autoantibodies induced by IFN in our study (anti-thyroid autoantibodies, ANCA) are associated with clinical symptoms.

Variations of cytokine levels and production in CAPD patients.

Macrophages, predominant cells in dialysates of patients on CAPD without peritonitis, produce a wide variety of substances including cytokines. The aim of this study was to examine the cytokine production in five uninfected patients. This work investigated the presence in dialysates of interleukin-1beta, interleukin-6, interleukin-8, tumor necrosis factor alpha and the ability of peritoneal macrophages to produce these cytokines. These results were compared with values obtained from control group in non-uremic conditions (peritoneal lavage with isotonic saline or dialysis fluid). All cytokines were detectable in dialysates. Interindividual variations in cytokine concentration in dialysates were wider than variations of production of cytokines ex vivo by stimulated and unstimulated cells. In control group, dialysis fluid inhibited the cytokine production and with isotonic saline, cells produced less cytokines than dialysis patients' cells. The highest levels of interleukin-1 and tumor necrosis factor in dialysates and the highest capacity to respond to LPS were observed in patients having the shortest duration of dialysis. The variability observed did not seem to be due to cells themselves but to their environment.

[Cold agglutinins and cryoglobulinemia in a patient with hepatitis C]. / Agglutinines froides et cryoglobulinémie chez un patient avec une hépatite C.

OBJECTIVES: Cold agglutinins and cryoglobulins are uncommon in the same patient as observed in our case. CASE REPORT: A 74-year-old patient suffered repeated episodes of hemolytic anemia for one year and had hepatitis C anti-virus antibodies. Mixed cryoglobulinemia was found at levels which increased during episodes of acute hemolysis in addition to anti-I cold agglutinins. Two-dimensional electrophoresis revealed identical oligoclonal cold agglutinins and cryoglobulins. DISCUSSION: Unlike mixed cryoglobulinemia, cold agglutinins are not known to occur subsequent to hepatitis C infection. The identical immunoglobulins observed in our patient suggest a common origin. Chronic anti-I cold oligoclonal agglutinins are rarely observed and could be an intermediary step towards monoclonal lymphopathy as has been described in prolonged hepatitis C infection.

[Detection and analysis of cryoglobulins: comparative study of a population of patients and one of healthy controls]. / Détection et analyse des cryoglobulines: étude comparative d'une population de malades et d'une population de témoins sains.

During an 11-month period, 270 cryoglobulins were identified in 735 sera from patients versus 34 in 82 sera from apparently healthy blood donors. Western-blot analysis of the cryoglobulins was performed and cryoglobulin protein content was determined using a colorimetric assay. Comparison of findings in the two groups suggested that a substantial proportion of cryoglobulins identified in the sera of patients are quantitatively and qualitatively indistinguishable from cryoglobulins identified in healthy controls.

Complement activation during low-density lipoprotein apheresis.

Complement system activation was investigated in two girls with familial homozygous hypercholesterolemia undergoing two monthly sessions on LA15 or LA40 (Kaneka liposorber). We determined blood levels of C3c and C3a, leukocyte counts, and plasma levels of C3c and C3a in the extracorporeal circulation device at the start of the sessions and 15 and either 60 or 120 min into them. Sequential eluates were collected from LA40 at the end of the sessions (0.5M NaCl, 1M hydroxylamine). Anaphylatoxin C3a increased throughout, especially with LA40. As previously reported, C3a was trapped in the dextran column but was noticeably present in efferent plasma. Besides many proteins, nonnative complement fragments bearing C3a and C3d antigens were detected in almost all the eluates, suggesting possible in situ complement activation. Practically, complement activation induced by the first filter is a risk; long-term side effects may arise from this extracorporeal circulation device.

[Mixed cryoglobulins and heterogeneity restriction]. / Cryoglobulines mixtes et restriction d'hétérogénéité.

Cryoglobulins from 143 patients were analyzed using an immunoblot technique with a nitrocellulose membrane. Fifty-four p. cent of cases were classified as type III, with polyclonal IgG, IgA, and IgM being the most common components. Type II was found in 24% of cases, with a monoclonal IgM in most instances. Lastly, classification was impossible in 22% of cases, usually because of the presence of oligoclonal IgM bands. Use of this highly sensitive technique to analyse cryoglobulins raises the problem of identification of patterns that are difficult to include in types II and III.

Leukotriene B4 in hemodialysis.

Leukotrienes are eicosanoids arising from arachidonic acid via 5 lipooxygenase, an enzyme essentially present in leukocyte cells. Leukotriene B4 might be an indicator of neutropolymorphonuclear leukocyte activation when there is contact with artificial membranes. The level of plasmatic leukotriene B4 was measured at three different times during the hemodialysis treatment in several patients undergoing dialysis on three different membranes (one cellulosic and two synthetics). A moderate increase of leukotriene B4 was observed early (at 15 min), comparable among the three membranes, but levels returned to baseline at 180 min. Leukotriene B4 production proved leukocyte activation and was probably related to a direct interaction with dialysis membrane. Nevertheless, complement intervention could not be excluded. Leukotriene B4 is one molecule more among the group of inflammatory mediators produced during hemodialysis treatment.

Immunohistochemical evaluation of reverse transcriptase in breast carcinoma with polyclonal antibodies raised in rabbit.

In this study, the presence of reverse transcriptase in breast tumours was examined with immunoperoxidase staining using antibodies raised in rabbit against reverse transcriptase of Moloney murine leukemia virus and against reverse transcriptase of avian myeloblastosis virus. The specificity of such antibodies was investigated with ELISA and Western blotting techniques. Five cases of infiltrating ductal carcinomas were found positive with the immune serum anti-reverse transcriptase of Moloney murine leukemia virus on 28 studied infiltrating ductal carcinomas, 2 infiltrating lobular carcinomas and 2 fibroadenomas.

[The kidney in Refsum's disease. Clinical, histologic and ultrastructural study of a case]. / Le rein dans la maladie de Refsum. Etude clinique, histologique et ultrastructurale d'une observation.

A renal biopsy was performed in a 41 year old man with Refsum's disease, following the onset of renal failure. 30% of the glomeruli were sclerosed, the others appeared normal. Ultrastructural examination revealed several types of inclusions within the tubular epithelial cells: dense bodies, simple or complex lipidic vacuoles and particularly structures composed of quadrangular microtubules with sides measuring 400 angstrom, rarely encountered in renal pathology. These dense elements, devoid of membrane and often in close contact with lipidic vacuoles, are found within the epithelial cells of the distal convoluted tubules and Henle's loops. They were localized extramitochondrially and no specimen was suggestive of a degraded form of the mitochondria. Some analogy can be made with the renal inclusions observed in Gaucher's disease.

[Evaluation of the biocompatibility of a new cellulose membrane by studying the complement system and histamine liberation]. / Evaluation de la biocompatibilité d'une nouvelle membrane cellulosique par l'étude du système du complément et de l'histaminolibération.

This biocompatibility of the new cellulosic membrane hemophane (HE) is compared to that of cuprophane (CU) in ten maintenance hemodialysis (HD) patients dialyzed on the two types of membranes in randomized order, under otherwise similar technical conditions. Total white blood cell (WBC) and differential counts, blood concentrations of C3a, and C3d and histamine are determined at start of dialysis (TO) and 10, 20 and 180 minutes thereafter. HE is distinct from CU in exerting a minor effect on the generation of C3a, a minor drop of leucocytes during the course of dialysis (P less than 0.01) and also by a lesser increase in blood histamine concentration (P less than 0.05). Histamine liberation is observed on CU together with the generation of anaphylotoxin C3 and with a diminution of circulating basopolymorpho-nuclear cells. According to the variations observed for these three parameters (C3a, leucocytosis and blood histamine concentration), HE appears as being more biocompatible than CU.

Determination of serum neuron-specific enolase by differential immunocapture.

A new method for the determination of serum neuron-specific enolase is presented. It consists of two steps: first, an immunocapture of gamma-subunit containing isoenzymes by absorption on immobilized anti-gamma antibodies; second, bioluminescence assay of enolase activities in untreated control samples and in the supernates of antibody treated samples. Total and alpha alpha activities are obtained, from which the neuron-specific enolase activity (alpha gamma + gamma gamma) can then be calculated by difference. As compared to the procedures currently in use, the immunocapture method is very rapid (30 min) and is more suitable for small series of determinations as needed in clinical chemistry applications. Reference interval values for serum found by this method agree with published data. When tested with samples from patients suffering from neuroblastoma or small cell lung cancer, it confirms the specific elevations in neuron-specific enolase activity previously described for these cancers, using other analytical approaches.

IgG purification to measure the level of an iodinated thyroglobulin peptide, the 3,5,3',5' tetraiodo-l-tyrosyl-l-tyrosine in human serum.

Antibodies reacting with 3,5,3',5' tetraiodo-l-tyrosyl-l-tyrosine (I2Tyr-I2Tyr) were elicited in rabbits by immunization with an oxidized yeast conjugate coupled with I2Tyr-I2Tyr. Ion-exchange chromatography was used to purify immunoglobulins, in order to improve the specificity in measurement of I2Tyr-I2Tyr level in patient serum. IgG binding capacity versus I2Tyr-I2Tyr was considerably increased after immunoglobulin purification.

Histochemical study of epididymal secretions in the lizard, Lacerta vivipara. Localization of lectin-binding sites.

The epididymis of lizards elaborates voluminous secretory granules made of a central core and a peripheral vacuole which in the species Lacerta vivipara contain respectively an insoluble protein (protein H) and a soluble protein (protein L). After their discharge these secretions mix with spermatozoa. In order to detect the presence of carbohydrates in these secretions, lectins isolated from Canavalia ensiformis (con A) and from eleven other plants (lentil, soja, pea, gorse and several mushrooms), conjugated to fluorescein isothiocyanate, have been utilized in light-microscopic histochemical investigations of frozen sections from Lacerta vivipara epididymis. Whereas lectins having affinity for L-fucose, lactose, D-galactose and N-acetyl-D-galactosamine bound to central cores, lectins having affinity to D-glucose, N-acetylglucosamine and chitobiose bound to the peripheral vacuole. D-mannose or D-glucose seem to be present both in central cores and in peripheral vacuoles.

[Respiratory manifestations in hereditary angioneurotic edema]. / Manifestations respiratoires des oedèmes angioneurotiques héréditaires.

Hereditary angioneurotic oedema is an autosomal dominant state associated with a quantitative, and sometimes purely functional, deficiency of C1 esterase inhibitor (C1 INH). The clinical manifestations may begin during adulthood or childhood; they are periodical and of varying severity. Beside oedema of the skin and digestive disorders, respiratory disorders are bound to attract attention. They consist of laryngeal oedema, which may end in lethal asphyxia if tracheotomy is not performed, or, exceptionally, of pulmonary oedema requiring assisted ventilation, as in the case reported here. The diagnosis, suspected in the presence of a decrease in CH50 and C4, is confirmed by a quantitative assay of C1 INH, which is low, and/or by the Fong and Good's functional tests. The physiopathological mechanisms are complex. They involve complement activation through the classical route, and activation of the coagulation system contact phase. Patients with severe attacks now benefit from treatment with preparations of C1 INH in high concentrations. The best treatment, however, is prophylactic, using testosterone derivatives, danazol and stanozolol, which can be prescribed for long periods taking into account their usually moderate side-effects.