Stress conditioning in mice: alterations in immunity and tumor growth

The neuroendocrine and autonomic nervous systems are known regulators of brain-immune interaction. However, the functional significance of this interaction under stress is not fully understood. We investigated the effect of a stress paradigm by applying electric foot shock followed by three reminders, on behavior, immune parameters, and lymphoma tumor growth. Male C3H mice were divided into two groups: Group 1-exposed to electric foot shock followed by three reminders, and Group 2-untreated (controls). Sets of mice underwent the elevated plus maze, staircase, and hot plate tests. After foot shock, natural killer (NK) cell activity, and lymphocyte proliferation were measured. In addition, sets of mice were either vaccinated twice with B-cell lymphoma 38C-13 immunoglobulin for determination of anti-idiotype (Id) antibodies in sera, or inoculated with tumor cells and monitored for tumor development and survival time. Mice exposed to electric foot shock followed by the three reminders had higher NK cell activity, levels of anti-Id antibodies, and a higher proliferation rate of splenocytes in response to mitogens, than the control mice. The exposed mice also showed attenuated tumor growth. Thus, the stress paradigm inhibited tumor development and lead to some immune changes that were not accompanied by behavioral changes.     

Differential effects of physical and psychological stressors on immune functions of rats

To study the effects of different types or durations of stressors on immune functions, male Fischer rats were exposed to chronic physical (electric foot shock) or psychological (non-foot shock) stress induced in the communication box. Superoxide production by alveolar macrophages (AMs), mitogen-induced splenic lymphocyte proliferation, and splenic natural killer (NK) cell cytolysis were examined in vitro. Repeated exposure to physical stress suppressed superoxide production by AMs (-58%, p<0.05 for opsonized zymosan (OZ) and -51%, p<0.05 for phorbol 12-myristate 13-acetate (PMA)), although psychological stress suppressed superoxide production after 24 h of repeated exposures (-40%, p<0.05 for OZ and -47%, p<0.05 for PMA). Acute suppression of the blastic response of splenic lymphocytes was only found in the physical stress group (p<0.05), although the chronic effects were only found in the psychological stress group (p<0.05). NK cell activity was suppressed immediately after the acute physical stress (-30%, p<0.05), but no effects were found in the psychological stress group. These results underline the importance of distinguishing between physical versus psychological stressors when examining the effects of stress on immune functions.     

Fighting experiences and natural killer cell activity in male laboratory mice

There is currently much interest in the potential impact of psychological factors on immune responses. An attempt was made to assess the effects of dominant/subordinate polarity in male mice on the cytotoxic activities of their natural-killer (NK) cells. On the basis of repeated agonistic encounters, categories of subordinate and dominat animals were selected. These animals were compared with manipulated controls (introduced to a novel test cage without an opponent on each occasion) and undisturbed controls (who remained in isolation without manipulation). In terms of NK activity, immunosuppression was observed in both dominant and subordinate categories when compared to undisturbed controls. There were, however, no differences between the fighting exposed subjects and the manipulated controls, suggesting that stress accounts for any changes. © 1994 Wiley-Liss, Inc.     

Decline of natural killer cell target binding and lytic activity in mice exposed to rotation stress

We examined the effect of rotation-induced stress on the percentage distribution of NK-YAC-1 target-binding cells and natural killer (NK) cell activity from splenic lymphocytes of C3H/HeJ mice. Following a 6-day stress regimen, we observed a marked decline in both the percentage of target-binding cells and in NK cell activity. This decline was first evident 13 days after initiation of stress and persisted for 2 weeks. Our data indicate that intermittent rotation stress over a 6-day period results in a delayed but persistent deleterious effect on NK-YAC-1 target binding and NK cell activity.     

Time-of-day effects on response of natural killer cells to acute stress in men and women.

Diurnal influences on natural killer (NK) cell changes to acute stress were assessed in 21 men and 21 women assigned to either an acute stress (mental arithmetic) or control task condition. Sessions began at either 8 a.m. or 2 p.m. Number of NK (CD3-CD56+) cells and NK activity were measured at baseline, during the 5-min task, and 60 and 90 min after the task. Both morning and afternoon stress participants had elevated NK cell numbers during the task. After the task, number of NK cells decreased in morning stress participants but remained significantly above baseline levels 60 and 90 min posttask. NK cell numbers in afternoon stress participants decreased to below baseline levels 60 and 90 min after the task. Changes in NK activity were driven primarily by diurnal influences. NK activity increased in all morning participants and stayed increased 60 and 90 min posttask. NK activity of all afternoon participants also increased during the task but dropped below baseline 60 and 90 min later. Greater increases in NK levels and activity during the task were associated with greater heart rate changes.     

Sense of coherence moderates the relationship between life stress and natural killer cell activity in healthy older adults

A sense of coherence (SOC) has been found to be a strong predictor of health outcomes and life satisfaction in older adults. This study investigated mood and immune effects of anticipated voluntary housing relocation in 30 healthy older adults and 28 age-matched controls and examined whether SOC would buffer effects of relocation on natural killer (NK) cell activity. Movers completed assessments and had blood drawn 1 month before relocation to congregate living facilities; controls were assessed concurrently. Compared with the control group, movers showed decreased positive mood and NK activity and elevated thought intrusion. Positive mood mediated the relationship of moving with NK activity, whereas SOC moderated this relationship. Low SOC movers had the poorest NK activity; that of high SOC movers was less compromised. These findings are consistent with possible salutogenic contributions of SOC and positive mood to immune function in older adults facing stressful life transitions.     

Tolerance to drug-induced (poly I:C) natural killer cell activation: congruence with a Pavlovian conditioning model

Two experiments were conducted to assess the role of conditioning factors on the tolerance of drug-induced natural killer (NK) cell activity. A protocol in which mice were given four weekly injections of the immunostimulatory synthetic polynucleotide (poly I:C) paired with a complex environmental stimulus produced a reliable tolerance effect. The sensitivity of the observed tolerance to known decremental conditioning procedures--extinction (Experiment 1) and preexposure to the conditioning stimulus (latent inhibition, Experiment 2)--was investigated. The results indicated that posttreatment exposure to drug-signaling cues (i.e., extinction) significantly reversed tolerance such that NK cell activity was at a level comparable to that for controls receiving the drug for the first time (Experiment 1). In a similar fashion, pretreatment exposure to the drug-signaling cues (CS preexposure) inhibited the development of tolerance (Experiment 2). These results provide initial evidence that nonpharmacological manipulations of drug-signaling environmental cues can affect tolerance to the immunostimulatory effect of poly I:C, as would be expected with a conditioning analysis of such tolerance.     

Psychosocial predictors of natural killer cell mobilization during marital conflict.

This study examined how specific emotions relate to autonomic nervous and immune system parameters and whether cynical hostility moderates this relationship. Forty-one married couples participated in a 15-min discussion about a marital problem. Observers recorded spouses' emotional expressions during the discussion, and cardiovascular, neuroendocrine, and immunologic parameters were assessed throughout the laboratory session. Among men high in cynical hostility, anger displayed during the conflict was associated with greater elevations in systolic and diastolic blood pressure, cortisol, and increases in natural killer cell numbers and cytotoxicity. Among men low in cynical hostility, anger was associated with smaller increases in heart rate and natural killer cell cytotoxicity. These findings suggest that models describing the impact of stress on physiology should be refined to reflect the joint contribution of situational and dispositional variables.     

Neurotoxic lesion of the rostral perirhinal cortex blocks stress-induced exploratory behavioral changes in male rats

Exposure to stress leads to adaptive responses including both behavioral and physiological changes. This process is induced by the activation of multiple brain regions. The present study examined the role of the rostral perirhinal cortex (rPRh) in behavioral changes following electrical foot shock-induced stress. The rPRh of rats was lesioned bilaterally by local microinjections of 10 microg N-methyl-D-aspartic acid (NMDA) before foot shocks (0.7 mA, 1 s). The effects of these lesions on foot shock-induced changes in exploratory behaviors were tested in the open field (4 h, 48 h, 72 h, and 14 days after foot shocks) and the light-dark box (7 days after foot shocks). Foot-shocked and sham-lesioned rats showed several well known behavioral changes in the open field (e.g., immobility, reduction of exploratory activity) most marked at 48 h after foot shocks, and the light-dark box (e.g., reduction of time spent and activity in the lit compartment). All these stress-induced behavioral changes were blocked by neurotoxic lesions of the rPRh. Furthermore, rPRh lesions did not affect behavior in the open field and the light-dark box in unstressed rats. Taken together, these data indicate that the rPRh is involved in neurophysiological mechanisms that mediate changes induced by foot-shock stress in exploratory behaviors which indicate unconditioned fear or anxiety.     

Representation of the serial killer on the Italian Internet.

The representation of serial killers was examined from the analysis of 317 Web pages in the Italian language to study how the psychological profiles of serial killers are described on the Italian Internet. The correspondence analysis of the content of these Web pages shows that in Italy the serial killer is associated with words such as "monster" and "horror," which suggest and imply psychological perversion and aberrant acts. These traits are peculiar for the Italian scenario.     

NK(3) receptor agonism promotes episodic-like memory in mice

The mammalian tachykinins are a family of closely related peptides including substance P, neurokinin A, neurokinin B and, recently, also hemokinin-1. They are present in the peripheral and central nervous systems, and bind to three known neurokinin (NK) receptors, the NK(1)-, NK(2)- and NK(3) receptors. In both rodents and humans, NK(3) receptors are expressed in brain structures which have been associated with learning and memory. Evidence for a role of NK(3) receptors in learning and memory has been found in NK(3) receptor knockout mice. Here, we investigated the influence of the NK(3) receptor agonist, senktide (0.1, 0.2 and 0.4 mg/kg), on the performance of C57BL/6 mice in a recently developed episodic-like memory task. Since a promnestic effect of senktide was expected, we employed an experimental protocol that provided sub-optimal learning conditions for episodic-like memory. The results indicate that senktide promotes episodic-like memory in mice in a dose-dependent manner, providing, for the first time, evidence for an involvement of NK(3) receptors in episodic-like memory.     

Tolerance of pain as a measure of fear

Fear thresholds were measured in four experiments by exposing rats to electric shock in order to determine the maximal intensity rats would tolerate rather than enter a fear-arousing box and/or stop freezing. Increasing fear raised these thresholds. They were greater for rats having to escape shock to a fear-arousing box than for rats having to escape shock and fear to a neutral box. The forgetting functions for the latter two groups differed: the first group yielded a monotonic decay function, whereas the second group yielded an inverted U-shaped function. These thresholds decreased as a function of an avoidance learning procedure. Rats that had to escape shock to a fear arousing box did not do so immediately, although they had stopped freezing. An avoidance-avoidance conflict explanation for immobility was not found to be valid. A theoretical formulation based on the following two hypotheses was suggested to explain these results: the fear-aroused freezing (immobility) is an unlearned response; finding a way to escape the source of fear starts another unlearned response, withdrawal.     

Prognostic risk assessment in primary breast cancer by behavioral and immunological parameters

Although findings from recent animal studies suggest that behavioral factors such as "helplessness" play a role in cancer progression, very few such studies with humans have been carried out. The study investigated the predictive power of an immunologic effector cell, the natural killer (NK) cell, as well as selected psychological and demographic factors, to breast cancer prognostic risk status. It was found that NK activity predicted the status of cancer spread to the axillary lymph nodes. Patients who had low levels of NK activity were rated as well-adjusted to their illness; patients who had higher NK activity appeared to be distressed or maladjusted. These findings are discussed in the light of recent animal findings linking environmental stress and behavioral responsiveness to biological vulnerability via endocrine and immune pathways.     

Genetic differences in avoidance learning by Rattus norvegicus: escape/avoidance responding, sensitivity to electric shock, discrimination learning, and open-field behavior

The behaviors of rats selectively bred for either good or poor shuttle box avoidance learning were studied. The results of Experiment 1 indicated that the phenotypic difference in avoidance learning is not associated with differences in speed of escape or avoidance responding. Differences between the lines in frequency of intertrial responses (ITRs), which appear during training but not during pretest, suggest that ITRs in animals of the low-avoidance (SLA) line are more suppressed by electric shock than in animals of the high-avoidance (SHA) line. This result suggests that SLA animals may be more emotionally responsive than SHA animals. Experiment 2 demonstrated that the animals of the two lines do not differ in absolute sensitivity to electric shock, and Experiment 3 showed that the poor performance of the SLA line is not due to an inability to learn. Experiment 3 also provided evidence which suggests that the poor avoidance learning by SLA animals is due to their emotional reactivity. Observations of open-field behavior in Experiment 4 are consistent with this hypothesis. The major consistent correlate of the phenotypic difference in avoidance learning is greater emotionality or emotional reactivity in SLA than in SHA animals.     

Maternal separation in early life impairs tumor immunity in adulthood in the F344 rat

Neonatal stress alters the hypothalamic-pituitary-adrenal (HPA) axis in rodents, such that, when these animals are exposed to stress as adults they hypersecrete corticosterone. Given that glucocorticoids are immunosuppressive, we examined the impact of maternal separation on HPA axis reactivity, natural killer (NK) cytotoxicity, and tumor growth in Fischer 344 rats following chronic restraint stress in adulthood. Pups underwent a chronic stress protocol whereby they were separated from their dams for 3 h on postnatal days 1-21. In adulthood, corticosterone responses were assessed following exposure to chronic (6 days for 10 h) restraint stress. Rats allocated to the chronic stress condition were inoculated with MADB106 tumor cells on day 4 of the restraint protocol. Blood was assessed for NK cytotoxicity on the final day of the chronic restraint protocol, and tumor colonization was assessed 3 weeks thereafter. Maternal separation impaired developmental weight gain (P < 0.05), depressed NK cytotoxicity (P < 0.05), and increased tumor colonization in the presence of chronic restraint stress in adulthood (P < 0.00 l). These findings occurred independently of circulating plasma corticosterone as only adult stress exposure potentiated corticosterone responses (P < 0.05). Our findings indicate that maternal separation and chronic stress can impair NK cytotoxicity and hence tumor immunity, but these effects are not directly mediated by perturbations in HPA axis function.     

Avoidance,induction, and the illusion of reinforcement

Environmental events that impact reproductive success may be called phylogenetically important events (PIEs). Some promote reproductive success, like mates and food; others threaten reproductive success, like predators and injury. Beneficial PIEs induce activities that enhance them, and detrimental PIEs induce activities that mitigate or avoid them. Free-operant avoidance relies on electric shock as a proxy for injury, a PIE. One theory takes avoidance behavior to be reinforced by its reducing shock rate. A more complete explanation is that avoidance both reduces shock rate and is induced by the PIEs it usually prevents. Shocks received act in concert with shock-rate reduction, in a feedback system. Four parametric data sets were analyzed to show that avoidance is induced by received shock rate according to power functions. Avoidance is not reinforced at all; avoidance is induced by its failures. Induction explains not only avoidance itself, but also phenomena unique to avoidance, like warmup and effects of unavoidable shock. Induction explains behavior more generally than reinforcement, because induction explains not only food-maintained operant and nonoperant behavior, but also shock-maintained behavior, including avoidance. Reinforcement fails to explain behavior when reinforcement is defined as strengthening by consequences. Induction erases the distinction between consequences and antecedents.     

Acceleration and suppression of rats' responding to avoid foot shock and tail shock

Signalled response-independent shocks were superimposed on rats' wheel-turn responding to avoid shock administered to their feet through a grid floor or to their tails through fixed electrodes. In Experiment I, a tone paired with response-independent foot shock increased responding in three of four rats; a tone paired with tail shock increased responding in only one of four rats and suppressed responding in two rats. In Experiment II, a tone presented randomly with respect to response-independent shock had no reliable effect on responding to avoid foot shock or tail shock. In Experiment III, tail shock and foot shock were compared in a within-subject design while the temporal pattern of responding during conditioned stimuli was recorded. Responding during the conditioned stimulus preceding foot shock was characterized by initial suppression of responding at tone onset, followed by increased responding just before response-independent shock. Responding was suppressed throughout the conditioned stimulus preceding tail shock. Foot shock elicited bursts of responding, but tail shock did not.     

Repressive Coping and Blood Measures of Disease Risk: Lipids and Endocrine and Immunological Responses to a Laboratory Stressor1

Relations between repressive coping and a variety of health‐related variahles including insulin, lipids, catecholamines, and cellular immune components, were investigated in a laboratory study of acute stress among a sample of healthy male college students (N ‐ 83). Compared to nonrepressors, at baseline, repressors had fewer numbers of circulating CD4 (T‐helper) cells, greater numbers of natural killer (NK) cells. lower high‐density lipoprotein (HDL), a higher total/HDL cholesterol ratio, and higher fasting insulin levels. In response to an acute laboratory stressor (Stroop Color Word Conflict Test). repressors demonstrated an attenuated increase in the number of circulating NK cells compared to nonrepressors. Confounds such as physical activity, age, and smoking were unrelated to the dependent measures.     

Aversive experiences are associated with a rapid and transient activation of ERKs in the rat hippocampus.

The extracellular signal-regulated kinases (ERKs) family of mitogen-activated protein kinases (MAPKs) has been shown to participate in memory formation. We recently found that a hippocampal ERK/MAPK cascade is required for memory formation of an inhibitory avoidance training in rats. Here we reported that this learning task is accompanied by a rapid increase in the phosphorylation of hippocampal p44 MAPK. A single mild foot shock produced a similar effect and three consecutive foot shocks provoked the activation of both p44 and p42 MAPKs. In contrast, a brief exposure to the training box or the habituation to an open field did not alter hippocampal ERK/MAPK levels. Together, these findings indicate that aversive behavioral experiences induced a rapid and transient activation of ERK/MAPKs in the hippocampus.     

Effects of acute and chronic stress and of genotype on oxotremorine-induced locomotor depression of mice

The locomotor behavior of unstressed and stressed mice of two inbred strains, DBA/2 and C57/BL6, was investigated. Animals were tested in a toggle-floor box apparatus, 30 min after saline or oxotremorine treatment (ip). A dose of oxotremorine that did not depress the activity of na?ve mice (0.01 mg/kg) was chosen. Stressed mice were injected 24 h after either a single 2-h stress session (acute stress) or the last of 14 daily stress sessions of tube restraining (chronic stress). Acute stress did not modify the depressant effect of oxotremorine on locomotor behavior in either strain. On the contrary, chronic stress induced a clear sensitization of DBA but not C57 mice to the depressant effect of oxotremorine. These findings show that chronic stress may result in modifications of the cholinergic function, and its behavioral correlates, and that these changes are modulated by the genetic makeup.