B cell early response gene expression coupled to B cell receptor,CD40 and interleukin-4 receptor co-stimulation: evidence for a role of the egr-2/krox20 transcription factor in B cell proliferation

B lymphocytes are activated following antigen stimulation of the B cell receptor but require co-stimulation with accessory molecules provided by interleukin (IL)-4/CD40 ligand for cell cycle progression and proliferation. By analyzing a panel of 11 early response genes induced by cross-linking of surface immunoglobulin, we show that CD40 signaling alone induces only 2 genes, c-myc together with an anonymous gene, 3L3, and that these are distinct from the set of genes induced in response to IL-4. Co-stimulation with the proliferative combination of anti-μ, IL-4 + CD40 signaling led to a fourfold enhancement of egr-2/krox20 expression over that seen with anti-μ alone. Egr-2 expression/activity was selectively inhibited by the immunosuppressive drug cyclosporin A, and antisense oligonucleotide blockade of Egr-2 activity elicited a dose-dependent inhibition of B cell proliferation. Taken together, these observations show that the early gene regulatory programs coupled to different surface receptors on B cells are largely distinct from each other, but that certain genes, exemplified by egr-2, may represent a point of convergence in the integration of different signaling pathways into the B cell proliferative response.     

Non-linearity of skin resistance response to intraneural electrical stimulation of sudomotor nerves

Intraneural electrical stimuli (0.3 mA, 0.2 ms) were delivered via a tungsten micro-electrode inserted into a cutaneous fascicle in the median nerve at the wrist in 16 normal subjects, and the effects on the sweat glands within the innervation zone were recorded as changes of skin resistance. In order to examine the relationship between the skin resistance level and the amplitude of transient resistance responses, trains of high frequency stimulation were used to reduce the skin resistance level and then transient resistance responses were evoked by single stimuli at 0.1 Hz. Regional anaesthesia of the brachial plexus in the axilla eliminated spontaneous sympathetic activity and reflex effects. At high skin resistance levels response amplitudes to single stimuli were low but they increased successively to a maximum at intermediate levels and then decreased again at low resistance levels. Repeated stimulation sequences evoked qualitatively similar response curves but quantitatively both response amplitudes and skin resistance levels were slightly reduced upon repetition. We suggest that the changes of response amplitudes are due to variable resistivity of the corneal layer. The shifts of the response curves with repetition of stimulation may result from increased hydration of the corneum. It is concluded that the variability of response amplitudes to constant stimuli makes the amplitude of a skin resistance response unsuitable as an indicator of the strength of sympathetic sudomotor nerve traffic.     

Ventilatory response to hypercapnia in sprint and long-distance swimmers

Summary Ventilatory response lines to carbon dioxide at rest were determined by the rebreathing method in 10 untrained subjects, 17 sprint swimmers, and 11 long-distance swimmers. It was found that the mean slope of the ventilatory response line of the swimmer was lower than that of the untrained group, and the mean slope of the long distance swimmer was lower as compared with the sprint swimmer, though these differences were statistically not significant. The differences in the hypercapnic drive between untrained subjects and swimmers obtained here is discussed in connection with their maximum oxygen uptake.     

Fast measurement of the CO2 partial pressure in gases and fluids

A CO₂-electrode system consisting of a membrane covered pH electrode, an electronic antilog modul and a special electronic analog circuit is described. Since the electrode output signal is a logarithmic function of the CO₂ partial pressure the output signal of the antilog module is proportional to the CO₂ partial pressure. The time course of the electrode signal has been analyzed after a step change of . This step response may be approximated by a sum of three exponential functions. Knowing the dynamic behaviour, the transfer function is formulated mathematically and a special analog circuit is constructed with a frequency response inverse to the frequency response of the electrode. Using this device the response time (T ₉₅) of the electrode system is diminished from 11,5 s to 750 ms after a step change of in gas (Luttmann, et al., 1974). If the time for the hydration of CO₂ is decreased by the addition of carbonic anhydrase the response time of the electrode is diminished to 6.5 s. Using the analog circuit yields a response time of 200 ms.Further studies were made to analyze the transient response in fluids at various flow velocities and various mountings. In order to analyze the influence of the fluid boundary layer on the surface of the electrode a photometric method has been developed (Luttmann and Mückenhoff, 1975), which allows to estimate the time course of the CO₂ partial pressure independently of and simultaneously with the electrode measurement.The experimental data are compared with a theory based on theoretical considerations of Schuler and Kreuzer (1967) and Crank (1956).List of Symbols A _i gain factor of thei-th compartment - A _i gain factor of the simulation network - C _i capacity - D diffusion coefficient - d electrode diameter - fraction of CO₂ - F _c (j) frequency response of the linearizing network - F _g (j) frequency response of the boundary layer - F _M (j) frequency response of the measuring system - F _M frequency response of the simulation network - I _i impedance transformer - j Gauss number (j ²=–1) - (j) frequency function of the CO₂ partial pressure - (t) time function of the CO₂ partial pressure - R _i electrical resistance - T _i time constant of the electrode - T _i time constant of the simulation network - T ₉₅ time for reaching 95% of the total difference - V voltage gain factor - v velocity of the streaming fluid - x coordinate - x(t) time function - X(j) corresponding frequency function - dilution factor - inverse time constant - thickness of boundary layer - ^* kinematic viscosity - thickness of diffusion layer - radian frequency Supported by the Deutsche Forschungsgemeinschaft, Sonderforschungsbereich 114 (Bionach)     

A role for the VLA-4 integrin in the activation of human memory B cells

It is generally recognized that activation through membrane effector molecules such as CD40 or the B cell receptor (BCR) is mandatory to allow B cells to proliferate and differentiate into antibody (Ab)-secreting cells in response to cytokines. We show here that purified tonsillar B cells can be stimulated directly by a cytokine combination to proliferate and secrete immunoglobulins when cultures are performed at high cell density. The contact-mediated activation of B cells in this experimental system is strongly inhibited both by anti-very late antigen (VLA)-4 monoclonal Ab and by a peptide containing the LDV sequence specifically recognized by the α4 integrin binding site. These reagents also significantly suppressed the B cell responses elicited by engagement of the BCR or CD40. Our data reveal that memory B cells but not virgin or germinal center B cells are sensitive to the direct stimulatory effect of cytokines in high-density cultures. Finally, we found that the dual expression of the α and β chains of VLA-4 is a distinctive feature of the memory B cell population. Collectively, our findings support the notion that VLA-4-dependent homotypic B cell interactions can mediate a co-stimulatory signal to human memory B cells and might participate in the B cell activation triggered through the BCR and CD40.     

Accuracy of eosinophils and eosinophil cationic protein to predict steroid improvement in asthma

BACKGROUND: There is a large variability in clinical response to corticosteroid treatment in patients with asthma. Several markers of inflammation like eosinophils and eosinophil cationic protein (ECP), as well as exhaled nitric oxide (NO), are good candidates to predict clinical response. AIM: We wanted to determine whether we could actually predict a favourable response to inhaled corticosteroids in individual patients. METHODS: One hundred and twenty patients with unstable asthma were treated with either prednisolone 30 mg/day, fluticasone propionate 1000 microg/day b.i.d. or fluticasone propionate 250 microg/day b.i.d., both via Diskhaler. They were treated during 2 weeks, in a double-blind, parallel group, double dummy design. We measured eosinophils and ECP in blood and sputum, and exhaled nitric oxide as inflammatory parameters before and after 2 weeks in order to predict the changes in forced expiratory volume in 1 s (FEV1), provocative concentration of methacholine causing a 20% fall in FEV1 (PC20 Mch), and asthma quality of life (QOL). Secondly, to test whether these results were applicable in clinical practice we determined the individual prediction of corticosteroid response. RESULTS: We found that changes in FEV1, PC20 Mch and QOL with corticosteroids were predominantly predicted by their respective baseline value and to a smaller extent by eosinophils in blood or sputum. ECP, measured in blood or sputum, was certainly not better than eosinophils in predicting clinical response to corticosteroids. Smoking status was an additional predictor for change in FEV1, but not for change in PC20 Mch or QOL. Prediction of a good clinical response was poor. For instance, high sputum eosinophils (> or = 3%) correctly predicted an improvement in PC20 Mch in only 65% of the patients. CONCLUSION: Our findings show that baseline values of the clinical parameters used as outcome parameters are the major predictors of clinical response to corticosteroids. Eosinophil percentage in blood or sputum adds to this, whereas ECP provides no additional information. Correct prediction of clinical response in an individual patient, however, remains poor with our currently used clinical and inflammatory parameters.     

A double-blind, placebo-controlled study of the effects of lithium on cognition in healthy subjects: mild and selective effects on learning

Background: Several studies have shown cognitive impairment in short-term memory, long-term memory and psychomotor speed in bipolar patients taking lithium. The aim of the study was to look at the effect of lithium in normal subjects (N=30) taking lithium for 3 weeks. A comprehensive battery was used to assess attention and memory. Methods: Subjects were randomized to double-blind treatment with either lithium (N=15) or placebo (N=15) for a 3-week period. Thirteen participants in the lithium group and 15 in the placebo group completed the study. The lithium and placebo were administered twice daily in doses varying from 1050 to 1950 mg (mean=1569 mg). The initial daily dose was calculated according to the Pepin formula to achieve a blood serum lithium level of about 0.8 mmol/l. Cognitive performance (attention, memory) was assessed in each subjects during three periods, i.e. at baseline, after 3 weeks of lithium or placebo, and 2 weeks after discontinuation of study medication. Results: In short-term memory tasks, the performance of subjects in the lithium group was worst 3 weeks after lithium treatment compared to 2 weeks after discontinuation. In long-term memory, a significantly higher number of words was recalled by the placebo group but not the lithium group. Conclusions. Lithium may have an effect on learning when long-term explicit memory test are administered repeatedly. It means that the practice effect when a subject performs the same task several times is less in the lithium-treated group than in the placebo group. This practice effect is related to the learning of a task.     

An Attempt to Elk it Cardiac Orienting and Defense Responses in the Newborn to Two Types of Facial Stimulation

Cardiac and behavioral reactivity of the human newborn to facial stimulation eliciting approach and escape responses were compared in order to test the distinction between cardiac orienting and defensive reactions. Each infant received 8 trials each of check stimulation (stroking near the mouth) and ear stimulation (pinch on the ear lobe). HR response to both tactile stimuli were accelerations of different amplitude when motor responses were also present. When no overt behavioral response was observed, stroking on the check elicited cardiac deceleration while ear stimulation again elicited acceleration. Thus, cardiac orienting was demonstrated in newborns when a rooting stimulus was presented that did not elicit overt head turning. The HR response to ear stimulation on trials unaccompanied by observed movements was a larger acceleration than to cheek stimulation when movement was present. This finding suggests that movement itself does not produce the observed HR increase, but rather that central processing of the signal value of the stimulus determines both overt and cardiac responding.     

The effect of the muscarinic antagonist scopolamine on regional cerebral blood flow during the performance of a memory task

Scopolamine, a muscarinic antagonist, impairs memory performance in both humans and animals. In this study, repeated measurements of regional cerebral blood flow (rCBF) were made in normal volunteers whilst performing auditory verbal memory tasks, before and after the administration of scopolamine (0.4 mg s.c.) or placebo. Compared to placebo, scopolamine increased blood flow in the lateral occipital cortex bilaterally and the left orbitofrontal region. Scopolamine decreased rCBF in the region of the right thalamus, the precuneus and the right and left lateral premotor areas. Scopolamine attenuated memory-task-induced increases of rCBF in the left and right prefrontal cortex and the right anterior cingulate region. These data suggest that acute blockade of cholinergic neurotransmission affects diverse brain areas, including components of the visual and motor systems, and, in addition, modulates memory task activations at distinct points in a distributed network for memory function.     

Quantitation of antigen-specific immune responses in human immunodeficiency virus (HIV)-infected individuals by limiting dilution analysis

The lymphocyte proliferative response to recall antigens is lost following HIV infection. We sought to devise a means by which the functional immune status of persons in the early stages of HIV infection could be monitored quantitatively. The response to tetanus toxoid was examined in 45 HIV-infected individuals and 11 controls using conventional lymphocyte proliferative assays concurrently with limiting dilution analysis utilizing the secretion of interleukin-2 as the measure of a response. Our data show that the limiting dilution analysis detects tetanus toxoid-reactive T cells in 80% of those tested, as compared to only 44% by proliferation. However, the frequency of tetanus-reactive T cells in HIV-infected individuals (median frequency = 1/59,156) is decrease five-fold as compared to seronegative controls (median frequency = 1/11,599). Longitudinal studies demonstrated a time-dependent decrease in the frequency of tetanus-specific T cell responses in the HIV-infected individuals. Thus, the limiting dilution analysis is a quantitative approach for detecting antigen-specific T cells in HIV-infected individuals, and may be used to monitor changes in T cell function in HIV infection.