Interferon-α activates cytotoxic function but inhibits interleukin-2-mediated proliferation and tumor necrosis factor-α secretion by immature human natural killer cells

Natural killer (NK) cells play an important role in host defense mechanisms against infection and neoplasia. Interferon- (IFN-) has been shown to activate NK cells and to augment their cytotoxic activity, albeit its role in the maturation pathway of NK cells has not been elucidated. The present study examined whether IFN- activates the immature NK subset (Free cells) to become cytotoxic and also ascertained whether IFN- uses the same pathway of activation as that mediated by interleukin-2 (IL-2). Incubation of sorted Free cells overnight with IFN- resulted in augmentation of their cytotoxic function against NK sensitive target cells. The enhanced cytotoxic activity was not accompanied by a new recruitment of NK-target binder cells but by an increase in the frequency of killer cells in the conjugate fraction. Activation of the Free subset by IFN- resulted in upregulation of CD69, CD11b, and CD2 surface expression and stimulated secretion of IFN-. Unlike IL-2, IFN- did not stimulate the Free cells to proliferate or secrete TNF- and activation of cytotoxicity and modulation of surface antigens by IFN- were independent of TNF-. The failure of IFN- to stimulate secretion and proliferation by Free cells appeared to be mediated by negative signals. This was corroborated in experiments demonstrating that when Free cells were cultured with both IFN- and IL-2, a significant inhibition was observed for both the IL-2 dependent secretion of TNF- and proliferation. These results demonstrate that IFN- serves as both an activator and a regulator of NK function. Further, activation of the immature Free NK cells by IL-2 and IFN- proceeds by TNF--dependent and independent pathways, respectively. The findings also support our contention that the mechanism of activation of the cytotoxic machinery of NK cells is not linked to the mechanism of activation of cytokine secretion and/or proliferation.Abbreviations used IFN interferon - IL interleukin - PBL peripheral blood leukocytes - PE phycoerythrin - PE-GAM PE-conjugated Fab2 goat anti-mouse IgG - NK natural killer - NRS normal rabbit serum - TNF tumor necrosis factor - FCS fetal calf serum - FITC fluorescein isothiocyanate - PBS phosphate-buffered saline - MACS magnetic cell sorting - ELISA enzyme-linked immunosorbent assay - BSA bovine serum albumin - PKC protein kinase C - mAb monoclonal antibody - PBMC peripheral blood mononuclear cells - BCLL B-chronic lymphocytic leukemia - E effector - T target     

Thermoregulatory responses to cold during a 7-week acclimatisation process in rabbits

A group of six rabbits reared at +20°C ambient temperature was adapted to moderate cold by housing for seven weeks at +10°C. Rectal and skin temperatures, metabolic heat production and respiratory evaporative heat loss were recorded continuously over 1 h for each animal on 3 days per week in the climatic chamber.There was no significant change either of rectal or of ear skin temperature during the acclimatisation process. On the other hand, metabolic heat production was progressively reduced (20% in the 7th week). Slight changes of mean skin temperature and respiratory evaporative heat loss could not account for compensation. Therefore it must be concluded that both adaptive improvement of peripheral insulation and reduction of heat production were achieved during the acclimatisation process. Both processes together ensure that deviations of core temperature are minimal. The possible origin of the functional adaptive effects is discussed. The results are in full agreement both with former neurophysiological results and with system-theoretical considerations of adaptive processes.Dedicated to the 60th birthday of Professor Dr. Kurt Brück     

Membrane attack complex of complement and 20 kDa homologous restriction factor (CD59) in myocardial infarction

In order to investigate the mechanism of deposition of the complement membrane attack complex (MAC) in cardiomyocytes in areas of human myocardial infarction, the 20 kDA homologous restriction factor of complement (HRF20; CD59) and complement components (C1q, C3d and MAC) were analysed immunohistochemically using specific antibodies. Myocardial tissues obtained at autopsy from nine patients who died of acute myocardial infarction were fixed in acetone and embedded in paraffin. The ages of the infarcts ranged from about 3.5 h to 12 days. In cases of myocardial infarction of 20 h or less, MAC deposition was shown in the infarcted cardiomyocytes without loss of HRF20. Where the duration was 4 days or more, the cardiomyocytes with MAC deposition in the infarcted areas also showed complete loss of HRF20. Outside the infarcts, HRF20 in the cardiomyocytes was well preserved without MAC deposition. The present study suggests that the initial MAC deposition in dead cardiomyocytes can occur as a result of degradation of plasma-membrane by a mechanism independent of complement-mediated injury to the membrane. Loss of HRF20 from dead cardiomyocytes may not be the initial cause of MAC deposition, but may accelerate the deposition process of MAC in later stages of infarction.     

Respiratory response to CO2 by controlled alveolar hypercapnia

A method of assessing the respiratory response to a hypercapnic stimulus after an increase in alveolar pCO₂ in accordance with an assigned program is suggested. The results are independent of the metabolic level, resistance to respiration, and other factors. Unlike the widely used rebreathing method, this new method enables the ventilatory sensitivity to CO₂ to be compared at rest, during muscular work, when the resistance to respiration is changed, and so on. It can also be used for both clinical and experimental investigations.Respiratory Physiology Group, I. P. Pavlov Institute of Physiology, Academy of Sciences of the USSR, Leningrad. (Presented by Academician of the Academy of Medical Sciences of the USSR V. N. Chernigovskii.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 89, No. 1, pp. 6–7, January, 1980.     

Inhibition of C5 or absence of C6 protects from sepsis mortality

Inhibiting complement anaphlytoxin C5a during sepsis may prevent sepsis mortality. Although human anti-C5 antibodies exist, their therapeutic use in microbial sepsis has been avoided because of the hypothesis that inhibiting C5b will prevent formation of the bactericidal membrane attack complex (MAC) and worsen clinical outcome. We wished to test the hypothesis that inhibition of C5 would improve outcomes in sepsis. Sepsis was induced in rats by laparotomy and cecal ligation and puncture (CLP) by an IACUC-approved protocol. Sham animals underwent laparotomy without CLP. Following CLP rats were randomized to receive a single IV dose of purified IgG ant-C5 antibody (Ab) or control IgG Ab. Anti-C5 Ab treated rats (n = 20) had significantly lower mortality vs. controls (n = 21), 20% vs. 52% (P = 0.019, log-rank). Analysis of bacterial load by culture of spleen and liver homogenates showed a reduction in colony forming units in anti-C5 Ab treated rats vs. control IgG (P = 0.003 and 0.009, respectively). Anti-C5 treatment reduced lung injury as measured by total MPO content of lung tissue (P = 0.024). Finally, rats genetically deficient in C6 production, unable to form MAC but capable of producing C5a and C5b, were protected from CLP-induced sepsis mortality. Our results show that in anti-C5 antibody therapy prevents CLP sepsis-induced mortality and improves lung injury. Inhibition of the complement MAC does not increase bacterial load or mortality, therefore, the use of anti-C5 therapy may be beneficial rather than detrimental in sepsis.     

The role of testosterone in the feedback control of male FSH and LH secretion

Summary Intramuscular administration of 250 mg testosterone oenanthate per week over a period of 21 weeks treatment rapidly and sustainedly suppressed serum LH as well as FSH levels in seven normal males, while serum testosterone rose by a factor of approximately two. These together with other data provide increasing evidence for a feedback control of FSH secretion by gonadal steroids in the male in addition to the already described but as yet undefined tubular testicular factor.     

Regulation and possible physiological role of the Ca2-dependent K+ channel of cortical collecting ducts of the rat

In the luminal membrane of rat cortical collecting duct (CCD) a big Ca²⁺-dependent and a small Ca²⁺-independent K⁺ channel have been described. Whereas the latter most likely is responsible for the K⁺ secretion in this nephron segment, the function of the large-conductance K⁺ channel is unknown. The regulation of this channel and its possible physiological role were examined with the conventional cell-free and the cell-attached nystatin patch-clamp techniques. Patch-clamp recordings were obtained from the luminal membrane of isolated perfused CCD segments and from freshly isolated CCD cells. Intracellular calcium was measured using the calcium-sensitive dye fura-2. The large-conductance K⁺ channel was strongly voltage- and calcium-dependent. At 3 mol/l cytosolic Ca²⁺ activity it was half-maximally activated. At 1 mmol/l it was neither regulated by cytosolic pH nor by ATP. At 1 mol/l Ca²⁺ activity the open probability (P _o) of this channel was pH-dependent. At pH 7.0 P _o was decreased to 4±2% (n=9) and at pH 8.5 it was increased to 425±52% (n=9) of the control. At this low Ca²⁺ activity the P _o of the channel was reduced by 1 mmol/l ATP to 8±4% (n=6). Cell swelling activated the large-conductance K⁺ channel (n=14) and hyperpolarized the membrane potential of the cells by 9±1 mV (n=23). Intracellular Ca²⁺ activity increased after hypotonic stress. This increase depended on the extracellular Ca²⁺ activity. A possible physiological function of the large-conductance K⁺ channel in rat CCD cells may be the reduction of the intracellular K⁺ concentration after cell swelling. Once this channel is activated by increases in the cytosolic Ca²⁺ activity it can be regulated by changes in cellular pH and ATP.Supported by DFG Schl 277/2-3     

Hyposmolarity-activated fluxes of taurine in astrocytes are mediated by diffusion

In order to obtain information about the mechanism responsible for swelling associated taurine release in astrocytes, the kinetics of taurine uptake in cultured astrocytes from mouse cerebral cortex was studied under isosmotic and hyposmotic (50% osmolarity) conditions. It was found that the V_max for the high affinity component of taurine uptake was unaffected by exposure of the astrocytes to hyposmotic conditions and that the K_m value was somewhat increased. Contrary to V_max, the non-saturable component of the uptake was greatly increased (2.5-fold) after exposure of the cells to hyposmotic media leading to cell swelling. In addition to the kinetic characterization of taurine uptake the actual intracellular taurine content after incubation (15 min) in isosmotic or hyposmotic media with different taurine concentrations (0–100 mM) under Na⁺-free conditions was determined. At taurine concentrations < 30 mM corresponding to the intracellular content in cells not exposed to taurine, exposure to hyposmotic media led to a decrease in the intracellular taurine content. At higher external taurine concentrations (> 30 mM) the intracellular taurine contents were dramatically increased after exposure to hyposmotic conditions. The increase in intracellular taurine seen under hyposmotic conditions at 100 mM external taurine could be significantly reduced by 100 μM DIDS (4,4′-diisothiocyanatostilbene-2,2′-disulfonate). Altogether these results suggest that a diffusional process rather than the high affinity taurine carrier is involved in the swelling induced increase in astrocytic taurine influx and efflux.     

Implantation, interception and contraception

The factors involved in post-fertilization events leading toimplantation in mammals are discussed with special referenceto potential forms of interception. The stages of embryonicgrowth until implantation are considered initially. The growthand differentiation of the uterine endometrium is then described,followed by the events occurring during the apposition and invasionof the implanting embryo. Several potential approaches to newforms of interception are considered, and the advantages anddisadvantages of each of them are evaluated. Among them, newvaccines against the zona pellucida, inactivation of the secretionsof the blastocyst, hatching, the activity of the pinopodes,and the endometrial proteins produced in the secretory phaseseem to offer various and varied targets. Some existing methodsof fertility regulation may act by affecting these stages ofdevelopment, e.g. RU486 may interfere with pinopod function.Various physiological and embryonic consequences of interferingwith these stages of pregnancy are discussed.     

LH syndrome and brain catecholamine levels after lesions of the nigrostriatal bundle

Lesions of the nigrostriatal bundle (NSB), whose fibers pass through the medial portions of the internal capsule and the immediately adjacent lateral hypothalamus (LH), produced a more severe aphagia, adipsia, and disturbance of water regulation than did lesions of the medial forebrain bundle (MFB). When deprived of food, animals with NSB lesions drank significantly less than controls and animals with MFB lesions. NSB lesions also produced greater decreases in telencephalic content of the catecholamines than MFB lesions, while the reverse was true for serotonin. Water intake during food deprivation was highly correlated with telencephalic catecholamine levels in animals with NSB lesions. Thus, the inability to regulate water intake in the absence of food, one of the characteristic and long lasting effects of the LH syndrome, appears to be due to destruction of the NSB and the consequent decline in telencephalic content of catecholamines.