Distribution of complement receptors on human normal and malignant mononuclear cells

Mononuclear cells from normal human subjects and patients with chronic lymphocytic leukemia (CLL), chronic lymphosarcoma cell leukemia (LCL), and hairy cell leukemia (HCL) were labeled with fluoresceinated, purified human C3b (FI-C3b) and analyzed using the fluorescence- activated cell sorter (FACS). FI-C3b labeled 17.6% +/- 6.0% of peripheral blood mononuclear cells (PBM) from 20 normal subjects, which, when separated by the FACS, consisted of B lymphocytes and approximately 5% monocytes. Analyses in which either monocytes or B lymphcoytes were excluded from consideration demonstrated that both these cell types were labeled by the FI-C3b with a heterogeneous distribution of fluorescence intensity, indicating either heterogeneity of CR density or variable avidity of individual CR for the FI-C3b. FACS profiles of PBM ( < 5% monocytes) from 14 of 15 patients with CLL showed a homogeneous distribution of very low fluorescence intensity, with > 60% of the cells being slightly more fluorescent than unlabeled controls. This low, homogeneous distribution of fluorescence is strikingly similar to profiles of CLL cells labeled with anti-Ig reagents and suggests homogeneity of low CR density and/or avidity. Similarly, CR+ mononuclear cells from five patients with HCL and three patients with LCL displayed more homogeneous FI-C3b labeling than normal CR+ PBM. Homogeneity of FI-C3b binding to CLL, LCL, and HCL cells further supports the concept for a clonal origin for these disorders.     

Significance of Ph1-negative marrow cells in Ph1-positive chronic granulocytic leukemia

Fifty-six of 195 Ph1-positive patients with chronic granulocytic leukemia were found to have Ph1-negative metaphases in marrow aspirates on one or more occasions. In 22 cases, Ph1-negative cells were found prior to initiation of antileukemic therapy. Five patients were in the blastic stage of the disease when Ph1-negative mitoses were seen. The finding of Ph1-negative cells appeared to be related principally to short duration of CGL and to administration of antileukemic therapy (conventional agents and doses, in most cases). Ph1-negative cells were usually not found more than 2 yr after the diagnosis of leukemia, but in a few cases, they were seen as long as 5-10 yr after diagnosis. Only a minority of metaphases analyzed were Ph1-negative, except in the case of 6 patients who transiently had 50% or more Ph1-negative cells after antileukemic therapy. The presence of Ph1-negative cells in marrow was not associated with any survival advantage in this series.     

P20 Red tattoo reactions

Tattoos are becoming increasingly popular, although reactions to tattoos remain relatively uncommon. We describe 4 patients with a variety of red tattoo reactions, one responding well to intralesional steroid therapy. Case 1: A 50‐year‐old man presented with a florid, inflammatory reaction confined to the red area of his forearm tattoo. Biopsy showed a dense lymphocytic and focal macrophage response to tattoo pigment. Mass spectrometry of biopsy tissue revealed high concentrations of titanium and iron. Patch testing was negative. Intralesional steroid injection has produced a marked improvement. Case 2: A 42‐year‐old man presented with an inflammatory reaction affecting the red area of his leg tattoo. Biopsy revealed a florid lymphoid reaction. Case 3: A 30‐year‐old man presented with an eczematous reaction within the red/brown pigmented areas of his tattoos, which was exacerbated by sun exposure. Patch testing showed a (+) positive reaction to cadmium after 96 hours. Photo patch testing was negative. The reaction settled spontaneously within 12 months. Case 4: A 37‐year‐old woman presented with a florid, indurated inflammatory reaction involving the red area of a shoulder tattoo. Patch testing revealed a (++) and (+) positive reaction to nickel and cobalt respectively with a doubtful (?+) reaction to mercury 0.5% in petrolatum after 96 hours. Tattoo reactions, especially red tattoo reactions can present with a spectrum of histological changes, including lichenoid, granulomatous, hypersensitivity, nodular, pseudolymphomatous or sarcoidal reactions. One of our cases responded well to intralesional steroid injection and one case resolved spontaneously.     

The differential effects of morphine, oxotremorine and antipsychotic drugs on DOPAC concentrations in rat brain

The effects of morphine and oxotremorine on concentrations of 3,4-dihydroxyphenylacetic acid (DOPAC) in the rat striatum and tuberculum olfactorium (TO) have been compared with the effects of the antipsychotic drugs haloperidol, chlorpromazine and clozapine. All the drugs elevated DOPAC concentrations in both brain regions. While the dose-response curves for the antipsychotic drugs were parallel, had steep slopes and similar maxima, the curves for morphine and oxotremorine were irregularly shaped but the curve for morphine in the TO had some similarity to that of the antipsychotic drugs. From these findings, it is concluded that the dose-dependent increase in striatal DOPAC effected by antipsychotic drugs can be used to differentiate them from other drugs known to elevate dopamine metabolites.     

Induction of craniofacial malformations in rhesus monkeys (Macaca mulatta) with cyclophosphamide.

Administration of cyclophosphamide to pregnant rhesus monkeys induces two syndromes of craniofacial malformations characterized by underdevelopment of the face of different pattern depending on the timing of treatment. The first is cleft lip with cleft palate and exophthalmos. The second is a craniofacial dysmorphia with marked underdevelopment of the midfacial bones, highly arched closed palate, and either meningoencephalocele or persistent anterior fontanel. Since cleft lip and palate are among the most common malformations in man, and several human syndromes involving abnormal craniofacial development have features in common with the dysmorphic monkeys, these induced anomalies may prove valuable in investigations on the pathogenesis of such malformations and as surgical models.     

P31Patch test reactions in the elderly

Objective:  To investigate whether application of allergic patch tests with transparent semi‐occlusive adhesive plastic foils yields higher test sensitivity than when tapes are used. To study whether such foils compared to tapes allow a shorter application time of tests.
Methods:  We applied different doses of budesonide printed on polyester squares and vehicle control squares to budesonide allergic subjects for 4 days. Each subject was tested with a set of tests both with a tape and a foil. We assessed all tests when they had been detached and additionally those applied with foils at earlier time points. All assessments were performed both visually and with a laser Doppler perfusion imaging technique.
Results:  Test sensitivity is higher with foil applications than when tapes are used and the perfusion is higher with the foils in many cases. The foils allow detachment of visually positive tests before 48 hours in some subjects, regardless of dose.
Conclusions:  Test applications with transparent semi‐occlusive adhesive plastic foils is sensitive and should be considered for application of patch tests when a short application time is important as when tests are carried out with occupationally hazardous allergens or when test substances containing allergens are expected to be irritating.     

Surgical correction of pectus excavatum deformity and hypomastia

Female patients occasionally present with major pectus excavatum and hypomastia. The aim of this study was to investigate the clinical outcome of female patients who had combined surgical correction of both deformities. Since 1990, 12 young female patients underwent correction using a modelling sternochondroplasty with osteosynthesis using Borrelly’s slide fastener-handle. After 1 year, the material was removed, and the breast implants were placed in the subpectoral plane. Our investigation was based on functional, morphological, aesthetic and psychological criteria. Despite nonsignificant pulmonary function tests (p < 0.05), we noted a subjective improvement of physical capacity during exercise following pectus excavatum repair. Correction of thorax deformity increased the sternovertebral distance by a mean of 3.2 cm (range 1.5–5.5 cm) thus treating mediastinal compression. As for aesthetic results, all patients were satisfied or very satisfied. The psychological benefit was considered as phenomenal after surgical treatment. Repair of pectus excavatum by sternochondroplasty combined with correction of hypomastia for female patients suffering from a double deformity is possible with only two different operations.     

Neutralizing and nonneutralizing monoclonal antibodies to the human granulocyte-macrophage colony-stimulating factor receptor alpha-chain

A panel of monoclonal antibodies was raised against the low-affinity human granulocyte-macrophage colony-stimulating factor (hGM-CSF) receptor alpha-chain expressed as recombinant protein on murine FDC-P1 cells. All the selected antibodies were of the IgG2A isotype and bound to protein A. They each recognized both native and recombinant receptors by indirect surface immunofluorescence and by immunoprecipitation. Several of the antibodies also recognized presumably denatured receptors as detected by immunoblotting of sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Three different epitopes on the extracellular domain of the GM-CSF receptor alpha-chain were defined by these antibodies, and two of the epitopes did not appear to be involved in binding hGM-CSF or in interactions with the beta-chain of the GM-CSF receptor that are required for high-affinity binding of GM-CSF. On the other hand, the epitope recognized by antibody 2B7-17-A appeared to be critically involved in the binding of GM-CSF because this antibody completely abrogated both high- and low- affinity binding of GM-CSF to native and recombinant receptors. Antibody 2B7-17-A had a relatively high affinity for the GM-CSF receptor alpha-chain (kd = 3 nmol/L) and slow dissociation kinetics (kd = 0.002 min-1). These properties made the 2B7-17-A antibody a potent inhibitor of hGM-CSF biologic action in several different bioassays, with a half-maximal inhibitory dose of about 6 nmol/L (1 microgram/mL). This antibody could prove useful in alleviating any pathologic states mediated by excess GM-CSF levels and in defining the domains of the GM- CSF receptor required for ligand binding.     

Defective delivery of iron to the developing red cell of the Belgrade laboratory rat

Erythroid cell iron and transferrin uptake and release was studied in the anemia of the Belgrade laboratory rat (gene symbol, b), an autosomal recessive trait characterized by hypochromia and hyperferrinemia. When reticulocyte-rich red cells were incubated in vitro with doubly (59Fe, 125I) labeled transferrin, b/b cells demonstrated a significantly higher uptake of transferrin (164% of control at 60 min), and a significantly lower uptake of iron (21% of control at 60 min) than control cells. These findings with b/b cells were simulated by sodium-fluoride-treated control cells, but not by trypsin-treated control cells. When reticulocytes exposed to doubly labeled transferrin were incubated in normal rat plasma, there was a substantial loss of 125I from both the b/b cells (mean 71%) and control cells (mean 49%), but only a loss of 59Fe from the b/b cells (mean 21%). These findings suggest a defect in the delivery of iron to the b/b reticulocyte, which is distal to the binding of transferrin to its cell surface receptor.