Induction of antibody responses to influenza virus in human lymphocyte cultures. I. Role of interleukin 2

The in vitro T cell-dependent antibody response of human lymphocytes to influenza virus X31 was used to study the role of T cell-derived lymphokines in antigen-specific responses. Supernatant from cultures of phytohaemagglutinin-stimulated, pooled human tonsil cells (PHA-MLR) was capable of replacing T cells and inducing T-depleted tonsil cells to secrete influenza-specific antibody. The T cell-replacing activity of PHA-MLR supernatant co-purified with interleukin 2 (IL 2) on Ultrogel AcA54 gel filtration and reversed phase-high performance liquid chromatography. PHA-MLR supernatant and IL 2 also enhanced B cell proliferation induced by anti-mu or Staphylococcal aureus strain Cowan I (SAC). A murine monoclonal antibody directed against the human IL 2 receptor (Mab 2A3) was used to completely block the enhancement of influenza-specific antibody production mediated by PHA-MLR supernatant, purified IL 2, and recombinant human IL 2. Mab 2A3 did not affect the T-independent B cell proliferation induced by anti-mu or SAC, but abrogated the enhancing effect of the PHA-MLR supernatant and IL 2 in this culture system. Immunofluorescence studies failed to demonstrate binding of Mab 2A3 to B cells activated by the X31 influenza virus and IL 2, or by SAC. By using Mab 2A3 to mask out IL 2 effects in the influenza-specific culture system, no other B cell differentiating activities were revealed in supernatants from lymphocytic cultures stimulated with a variety of mitogens. Thus, our results indicate that the production of influenza-specific antibodies by T-depleted human lymphocyte cultures is absolutely dependent on the presence of both antigen and IL 2.     

Biofuel anode based on d-glucose dehydrogenase,nicotinamide adenine dinucleotide and a modified electrode

A biofuel cell anode has been made from a modified graphite electrode and immobilized d-glucose dehydrogenase [β-d-glucose:NAD(P)⁺ 1-oxidoreductase, EC 1.1.1.4 7] so that energy could be drawn from the conversion of d-glucose to d-gluconic acid. An equivalent amount of dihydronicotinamide adenine dinucleotide (NADH) was formed from NAD⁺ and reduced the surface groups of the modified electrode. Reoxidationn of the latter produced the electrons necessary for a power output from the cell. Electrode modification was made by adsorption of N,N-dimethyl-7-amino 1,2-benzophenoxazinium onto the graphite. A current density of 0.2 mA cm^?2 at a cell voltage of ～0.8 V was obtained for more than 8 h with a simulated oxygen cathode. The internal resistance in the cell, in particular in the separator, appeared to be the main current-limiting factor.     

Characterization of the cell surface receptor for a multi-lineage colony-stimulating factor (CSF-2 alpha)

125I-Labeled colony-stimulating factor (CSF) 2 alpha (interleukin 3, multi-CSF, and mast cell growth factor) was used to characterize receptors specific for this lymphokine on the cell surface of the factor-dependent cell line FDC-P2. CSF-2 alpha binding to these cells was specific and saturable. Among a panel of lymphokines and growth factors, only unlabeled CSF-2 alpha was able to compete for the binding of 125I-labeled CSF-2 alpha to cells. Equilibrium binding studies revealed that CSF-2 alpha bound to 434 +/- 281 receptors/cell with a Ka of 8.7 +/- 3.9 X 10(9) M-1. Affinity cross-linking experiments with the homobifunctional cross-linking reagents disuccinimidyl suberate, disuccinimidyl tartrate, and dithiobis(succinimidyl propionate) produced a radiolabeled band of Mr = 97,000 on intact cells and in purified cell membranes, while an additional band of Mr = 138,000 was produced upon cross-linking to intact cells only. The relationship between these two bands is discussed. The results indicate that the receptor for CSF-2 alpha on FDC-P2 cells consists at a minimum of a subunit of Mr = 72,500.     

'Perforated patch recording' allows long-term monitoring of metabolite-induced electrical activity and voltage-dependent Ca2+ currents in pancreatic islet B cells

We describe the application of 'perforated patch recording' using the pore-forming antibiotic nystatin, to monitor the electrical activity and underlying ionic currents of rat and human pancreatic islet B cells. We demonstrate that glucose-induced electrical activity is seen even in single B cells during current-clamp recordings lasting hours 'L-type' Ca2+-channel currents can also be monitored over this period of time. This technique may prove useful in examining hormone and neurotransmitter modulation of electrical activity in B cells, while minimizing the effects of cytoplasmic 'wash-out'.     

Propranolol and serotonin removal in lung injury

Indicator dilution technique was used to study effects of reduced vascular volume or acute injury on removal of low doses of [3H]propranolol and [14C]serotonin (5-hydroxytryptamine, 5-HT) by perfused rabbit lung. Glass-bead (500 micron) embolization doubled pulmonary arterial pressure (Ppa) at flow rates of 20, 50, and 100 ml/min, decreased volume of distribution by approximately 50%, and increased pulmonary vascular resistance by at least 60%. Before embolization, (flow rate 20 ml/min) removal of [3H]propranolol and [14C] 5-HT was 89 +/- 2 and 75 +/- 5%, respectively, and was unaltered by changes in flow rate. However, after embolization, [3H]propranolol and [14C]5-HT removal decreased in a flow-dependent manner, reaching 28 +/- 4 and 1 +/- 3% (P less than 0.05), respectively, at a flow rate of 100 ml/min. When phorbol myristate acetate (PMA, 200 nM) was perfused (50 ml/min) through the lungs for 15 min, Ppa increased from 13 +/- 1 to 25 +/- 2 cmH2O (P less than 0.05), whereas [3H]propranolol removal decreased from 92 +/- 1 to 75 +/- 5% (P less than 0.05) and [14C]5-HT removal decreased from 73 +/- 3 to 46 +/- 8% (P less than 0.05). The PMA also caused vasoconstriction, which could be partially blocked by adding papaverine (500 microM) to the perfusion medium. Under the latter conditions, Ppa increased to 19 +/- 1 cmH2O and [3H]propranolol removal was unaffected. However, the combination of PMA and papaverine reduced [14C]5-HT removal from 64 +/- 4 to 19 +/- 3%.(ABSTRACT TRUNCATED AT 250 WORDS)     

Asbestos and lung cancer in Glasgow and the west of Scotland.

OBJECTIVE--To quantify the relation between lung cancer and exposure to asbestos in men in west Scotland and to estimate the proportion of lung cancer which may be attributed to exposure to asbestos. DESIGN--An ecological correlation study of the incidence of lung cancer in men and past asbestos exposure. The unit of analysis was the postcode sector. Correction was made for past cigarette smoking, air pollution, and deprivation. SETTING--The region covered by the west of Scotland cancer registry, containing 2.72 million people and including Glasgow and the lower reaches of the River Clyde, where shipbuilding was once a major industry. SUBJECTS--All men diagnosed with lung cancer between 1975 and 1984 whose residence at the time of registration was within the west of Scotland. MAIN OUTCOME MEASURE--The population attributable risk for asbestos related lung cancer. RESULTS--An estimated 5.7% (95% confidence interval 2.3 to 9.1%) of all lung cancers in men registered in the west of Scotland during the period 1975-84 were asbestos related, amounting to 1081 cases. CONCLUSIONS--A considerable proportion of cases of lung cancer in men in Glasgow and the west of Scotland from 1975 to 1984 were asbestos related. Most of these may not have been considered for compensation by the Department of Social Security. Given the very small annual number of recorded cases of asbestosis this condition is probably not a prerequisite for the development of asbestos related lung cancer. A heightened awareness of the increasing incidence of asbestos related neoplasms and their more thorough investigation are recommended.     

The interaction of behavioural state, heart rate and resistance index in the human fetus.

The association and possible interactions of fetal behavioural state, fetal heart rate and vascular resistance was studied in 23 healthy pregnant women between 36 and 40 weeks' gestation. Doppler flow velocity waveforms were obtained from fetal cerebral (anterior cerebral, internal carotid and basilar), descending aorta and umbilical arteries during fetal behavioural state 1F and repeated during 2F. The Resistance Index (RI) was used as a measure of vascular resistance. Decreased vascular resistance was observed in all vessels except the umbilical artery during fetal behavioural state 2F compared to 1F (P less than 0.001). A significant interaction was observed between fetal heart rate and fetal behavioural state on the RI of the vessels studied, with a greater negative slope for RI plotted against fetal heart rate in fetal behavioural state 2F (-0.00139) compared to 1F (0.00005) (P less than 0.001). We conclude that the transition from fetal behavioural state 1F to 2F is associated with a significant reduction in cerebral and systemic vascular resistance, with no apparent change in placental resistance. Furthermore, fetal behavioural state and fetal heart rate interact, demonstrating a stronger association of fetal heart rate and RI in fetal behavioural state 2F in keeping with an increase in baroreflex sensitivity at this time.     

Effect of fluid intake on renal function during exercise in the cold

The effect of an imposed drinking discipline versus ad libitum drinking was studied on 21 healthy, well-trained volunteers, during a continuous 4.5-h march at an altitude of 1,700 m and an ambient temperature of 0 degree C, SD 1. Group I (n = 13) was instructed to drink 250 ml of warmed, artificially sweetened fluid every 30 min, whereas group II (n = 8) drank plain water ad libitum. The median fluid intake in group I was significantly higher than in group II (P less than 0.0002). Serum urea and osmolality decreased during the march in group I (P less than 0.05; P less than 0.002, respectively) with no significant change in group II. In both groups, a similar increase in haemoglobin concentration concomitant with a reduction in calculated blood and plasma volume was observed after exercise and did not correlate with the state of hydration. Total urine volume, creatinine clearance, urea clearance and potassium excretion were significantly higher and urinary osmolality was lower in group I than in group II (P less than 0.05). These results reflect a state of extreme "voluntary dehydration" in the control group when no fluid intake was obligatory. Thus, during exercise in the cold, under conditions similar to those in this study, a fluid intake of 150 ml.h-1 should be maintained in order to keep a urinary flow of about 1 ml.kg-1.h-1 and to achieve a good state of hydration.