Age-related changes in neural volume and microstructure associated with interleukin-6 are ameliorated by a calorie-restricted diet in old rhesus monkeys.

Systemic levels of proinflammatory cytokines such as interleukin-6 (IL-6) increase in old age and may contribute to neural atrophy in humans. We investigated IL-6 associations with age in T1-weighted segments and microstructural diffusion indices using MRI in aged rhesus monkeys (Macaca mulatta). Further, we determined if long-term 30% calorie restriction (CR) reduced IL-6 and attenuated its association with lower tissue volume and density. Voxel-based morphometry (VBM) and diffusion-weighted voxelwise analyses were conducted. IL-6 was associated with less global gray and white matter (GM and WM), as well as smaller parietal and temporal GM volumes. Lower fractional anisotropy (FA) was associated with higher IL-6 levels along the corpus callosum and various cortical and subcortical tracts. Higher IL-6 concentrations across subjects were also associated with increased mean diffusivity (MD) throughout many brain regions, particularly in corpus callosum, cingulum, and parietal, frontal, and prefrontal areas. CR monkeys had significantly lower IL-6 and less associated atrophy. An IL-6xCR interaction across modalities also indicated that CR mitigated IL-6 related changes in several brain regions compared to controls. Peripheral IL-6 levels were correlated with atrophy in regions sensitive to aging, and this relationship was decreased by CR.

Maternal pregnancy weight gain and cord blood iron status are associated with eosinophilia in infancy.

OBJECTIVE: Allergic disease is multifactorial in origin. Because iron nutrition affects immune responses and maternal pregnancy weight gain impairs fetal iron delivery while increasing fetal demands for growth, the study examined maternal pregnancy weight gain, newborn iron status and an index of atopic disease, infant eosinophilia. STUDY DESIGN: Within a larger prospective study of healthy newborns at risk for developing iron deficiency anemia, umbilical cord iron indicators were compared to infant eosinophil counts. RESULT: Infants who developed eosinophilia exhibited higher cord reticulocyte-enriched zinc protoporphyrin/heme ratio, P<0.05 and fewer cord ferritin values in the highest (best) quartile, P<0.05. If cord ferritin was in the upper three quartiles, the negative predictive value for infant eosinophilia was 90%. High maternal pregnancy weight gain predicted infant eosinophil counts, P<0.04, and contributed to cord ferritin predicting eosinophilia, P<0.003. CONCLUSION: Poor fetal iron status may be an additional risk factor for infant eosinophilia.

Rapid cortisol and corticosteroid-binding globulin responses during pregnancy and after estrogen administration in the squirrel monkey.

Plasma cortisol and corticosteroid-binding globulin (CBG) levels were assessed in pregnant squirrel monkeys and in intact and castrated males after estrogen administration. Pregnant females showed a rapid and dramatic rise in cortisol and CBG during the first 8 weeks after conception. Estrogen treatment also caused marked elevations in cortisol and CBG. Cortisol levels increased significantly by 24 h after estrogen injection and remained elevated for 6 weeks of treatment, but a relatively greater rise in CBG resulted in a higher CBG/cortisol ratio. The data support prior research indicating that estrogen can simultaneously stimulate adrenal output and the compensatory binding of circulating cortisol by increased CBG synthesis. In addition, it appears that even in the absence of exogenous treatment, the pituitary-adrenal axis of male squirrel monkeys is stimulated by estrogen derived either from the testes or by the peripheral conversion of testosterone to estrogen.

Fluoxetine treatment alters leukocyte trafficking in the intrathecal compartment of the young primate.

To evaluate possible long-term effects of exposure to monoaminergic drugs, blood and cerebrospinal fluid (CSF) samples were collected from adolescent monkeys that had been treated with desipramine and fluoxetine in infancy. This evaluation focused on the number and type of leukocytes in CSF as a reflection of cell trafficking in the intrathecal compartment. Monkeys administered fluoxetine 2 years prior to the sample collection evinced significantly higher numbers of leukocytes in CSF than did either control or desipramine-treated subjects. The elevated cell count was accounted for primarily by increased numbers of CD4+ and CD8+ lymphocytes. The finding of higher cell numbers in CSF was replicated in a second sample from the fluoxetine-treated monkey obtained 1.5 years later. Because the cell profile in blood was unaffected by the prior drug treatments, these observations indicate a need for further study of serotonergic influences on regulation of the intrathecal compartment in the developing individual.

Cytokine profiles of stimulated blood lymphocytes in asthmatic and healthy adolescents across the school year.

T cell cytokines play an important role in mediating airway inflammation in asthma. The predominance of a Th2 cytokine profile, particularly interleukin (IL)-4 and IL-5, is associated with the pathogenesis and course of asthma. The aim of this study was to test the hypothesis that a stressful life event alters the pattern of cytokine release in asthmatic individuals. Thirteen healthy controls and 21 asthmatic adolescents gave blood samples three times over a semester: midsemester, during the week of final examinations, and 2-3 weeks after examinations. Interferon-gamma (IFN-gamma), IL-2, IL-4, and IL-5 were measured from supernatants of cells stimulated with PHA/PMA for 24 h. Cells from asthmatic subjects released significantly more IL-5 during the examination and postexamination periods, whereas cells from healthy controls released significantly more IL-2 during the midsemester and examination periods, thereby indicating a bias for a Th2-like pattern in asthmatics and a Th1-like pattern in healthy controls. IL-4 and IL-5 production showed a marked decrease during and after examinations in healthy controls, whereas this decline was absent in asthmatics. The ratios of IFN-gamma:IL-4 and IFN-gamma:IL-5 also revealed significant changes in the profile of cytokine release across the semester. These results indicate differential cytokine responses in asthmatics that may become pronounced during periods of cellular activation.

Interleukin-1 beta differentially affects interleukin-6 and soluble interleukin-6 receptor in the blood and central nervous system of the monkey.

Two studies were conducted to investigate whether behavioral and physiological responses induced by administration of interleukin-1 beta (IL-1 beta) were also associated with changes in interleukin-6 (IL-6) and soluble IL-6 receptor levels (sIL-6R). Following intravenous injection of rhIL-1 beta, blood and cerebrospinal fluid (CSF) samples were collected from juvenile rhesus monkeys. Marked increases in IL-6 levels were evident at 1 h in both blood and intrathecal compartments. IL-1 beta also induced significant elevations in the release of ACTH and cortisol into the blood stream, and following high doses, the monkeys evinced signs of sickness behavior. The second study characterized the IL-beta dose-response relationship showing that these physiological changes were most evident at doses between 0.5 microgram and 1.0 microgram IL-1/kg body weight. Soluble IL-6 receptor concentration was also increased, but only in plasma. There was no detectable sIL-6R in CSF. The large release of IL-6 into CSF suggests that some behavioral symptoms may be due to intrinsic changes in central nervous system activity concomitant with the alterations in peripheral physiology.

Psychological disturbance differentially alters CD4+ and CD8+ leukocytes in the blood and intrathecal compartments.

To evaluate cellular changes in the intrathecal compartment in response to psychological stress, cerebrospinal fluid (CSF) and peripheral blood (PB) samples were obtained from rhesus monkeys under baseline and challenge conditions. Juvenile monkeys separated from their social companions overnight had elevated cortisol, increased polymorphonuclear (PMN), and fewer CD4+ and CD8+ leukocytes in PB. In contrast, in CSF there were more CD4+ and fewer CD8+ leukocytes, raising the CD4/CD8 ratio. Dexamethasone given intramuscularly caused similar hematological changes; i.e. neutrophilia and lymphocytopenia with fewer CD4+ and CD8+ leukocytes in PB. However, it did not induce similar changes in CSF, indicating that the stress-related shift of CD4+ leukocytes in the intrathecal compartment involves physiological processes beyond adrenocortical steroids.

Immune consequences of stroke and cerebral palsy in adults.

The possibility that brain damage results in a sustained dysregulation of lymphocyte responsiveness to the lymphokine, interleukin-2 (IL-2), was investigated in individuals who had experienced a unilateral stroke in adulthood or who presented with spastic hemiparesis since childhood. Following verification of unilateral brain damage via neuromotor assessment, and determination of their health status, blood samples were obtained to evaluate a panel of immune measures. Soluble interleukin-2 receptor (sIL-2R) and lymphocyte proliferative and cytolytic responses in the subjects with stroke or cerebral palsy were compared to age- and gender-matched controls. In addition, lymphocyte populations were enumerated via flow cytometry, and lymphocyte cyclic AMP (cAMP) levels were determined. Circulating blood levels of sIL-2R were significantly elevated in all individuals that had experienced unilateral brain damage. Cytolytic activity also failed to be stimulated to the normal level by in vitro treatment of lymphocytes with IL-2. Further, lymphocytes from the stroke subjects proliferated significantly less after mitogen and IL-2 stimulation. These functional differences were not accounted for by an abnormal leukocyte profile, although phenotypic analyses revealed subtle differences in the natural killer cell subsets. Overall, the findings indicate that individuals with brain damage may not respond appropriately when immune activation is required. These immune differences appear to be a stable trait given that they were manifested after both perinatal and adult brain insult in otherwise healthy, independently living individuals.

Early rearing conditions alter immune responses in the developing infant primate.

The influence of early rearing conditions on immunologic development was investigated in infant monkeys. Lymphocyte proliferation, natural killer cell activity, and antibody responses to tetanus vaccination were compared in 30 rhesus monkeys reared under five different conditions. Lymphocyte responses to two mitogens (concanavalin A and pokeweed) were significantly increased in infants from disturbed rearing conditions compared with control infants that had been reared in an undisturbed manner by their mothers. The largest increases occurred in nursery-reared monkeys that had been fed Similac infant formula. The nursery-reared monkeys also tended to show lower natural killer cell activity, but there were no significant differences in response to vaccination. These findings support other research indicating that psychologic and nutritional aspects of the early rearing environment may have long-lasting effects on some, but not all, immune responses in the developing infant.

Quinolinic acid and lymphocyte subsets in the intrathecal compartment as biomarkers of SIV infection and simian AIDS.

Cerebrospinal fluid (CSF) samples were collected from monkeys infected with SIVmac251 (SIV) or HIV-1/SIVmac chimeric viruses (SHIV(HXBc2) and SHIV(89.6P)) to investigate quinolinic acid (QUIN) levels in the intrathecal compartment. CSF levels of QUIN were elevated in the SIV-infected monkeys, especially in animals with end-stage disease, and in those infected with pathogenic SHIV(89.6P), but not after infection with the nonpathogenic construct SHIV(HXBc2). QUIN elevations occurred in association with reduced CD4+ and increased CD8+ lymphocytes, cellular alterations that were more pronounced in CSF than in the blood. These findings support the view that the intrathecal compartment provides a unique window on viral infection, and are in keeping with the a priori prediction that QUIN increases primarily in response to more pathogenic viral strains.

Psychological disturbance alters thymic and adrenal hormone secretion in a parallel but independent manner.

Thymosin-alpha 1 (thymosin alpha 1) and cortisol levels were evaluated in juvenile squirrel monkeys to investigate the influence of psychological disturbance on thymic and adrenal hormone activity. Hormone levels were assessed in peripheral circulation following removal of monkeys from their social group to establish the time course of thymosin and cortisol alterations. Thymosin alpha 1 was significantly decreased after social separation in association with increased adrenocortical activity, especially during the first day after being housed alone. The temporal pattern suggested that both hormone systems are involved in the acute inhibition of functional immunity observed following this type of psychological disturbance. A second study verified that the decrement in thymosin alpha 1 levels was replicable and also sensitive to psychosocial factors that influence the level of induced disturbance. In addition, changes in thymosin secretion could be attenuated partially by pharmacological inhibition of the cortisol response and opiate hormone action. Nevertheless, the decrease in thymosin alpha 1 secretion did not appear to be a secondary consequence of adrenocortical secretion, and instead probably emanates from a general shift in neuroendocrine activity.

Cortisol responses under different housing conditions in female squirrel monkeys.

Adult female squirrel monkeys were housed in a group, in pairs or individually. Plasma levels of cortisol obtained under basal and stress conditions once weekly for four weeks were significantly lower in pair-housed females than in those living in a social group or individually. The increment in cortisol levels after stress (induced by handling and ether anesthesia) also was smaller in females housed in pairs. The cortisol values of the pair-housed females were positively correlated with those of their partners. Basal cortisol levels in the group-living females showed a significant rank-order correlation with dominance status. This indicated that social interactions in group-living animals can influence cortisol levels in a complex manner, either increasing or decreasing them. The relatively lower pituitary--adrenal activation when a single partner was present also indicated that the social environment can affect an individual's general level of arousal and subsequently alter the response to stressful stimuli.

Temporal and social factors influencing behavioral and hormonal responses to separation in mother and infant squirrel monkeys.

The behavioral and hormonal responses of mother and infant squirrel monkeys (Saimiri sciureus) were examined to assess temporal and environmental factors that influence the response to separation. In two experiments evaluating the effects of 1-, 3-, 6-, and 24-hr separations, it was found that signs of infant behavioral agitation decreased over time, whereas adrenocortical activation persisted or even increased. Moreover, two separation environments were shown to differentially affect behavioral and hormonal responses. Separated infants vocalized significantly more when their mothers were proximal than when isolated, but showed lower cortisol levels in the adjacent separation than in the total-isolation condition. These data indicate that the intensity of the infant's calling response cannot be used to predict internal state (as reflected by cortisol levels). Furthermore, vocalization rate is highly dependent upon contingent stimuli, such as the presence of maternal cues. Following separation, the mothers also showed elevated cortisol levels. However, both the magnitude and pattern of the response differed considerably from that of the infant.

Context-dependent behavioral effects of interleukin-1 in the rhesus monkey (Macaca mulatta).

The behavioral effects of recombinant human interleukin-1 alpha (IL-1) in rhesus monkeys (Macaca mulatta) were assessed in 3 experimental paradigms: (1) a testing situation in which an initial quiescent period was followed by a challenge designed to evoke agitation; (2) a novel environment with a social partner; and (3) a working memory-dependent nonspatial cognitive task. In the first two experiments we replicated our previous observations that a high IL-1 dose (25 micrograms) induces somnolence in a quiet setting within 1 h. A lower IL-1 dose (1 ng) did not have these sedative properties, but both IL-1 doses significantly reduced the number of vocalizations made by the monkeys. In contrast, when the monkeys were challenged through direct eye contact with a human experimenter, the 25 micrograms IL-1 dose significantly increased agonistic behavior. Finally, performance on a working memory-dependent task (delayed non-matching-to-sample) was unaffected by doses of IL-1 ranging from 1 to 25 micrograms, possibly because the monkeys were tested after learning the task rather than during the acquisition phase. These results demonstrate that high levels of IL-1 in peripheral circulation can have potent behavioral effects in the nonhuman primate, but that the nature of the influence will depend on the context in which the animal is evaluated. Manifestation of cytokine-induced 'sickness behavior' appears to require a permissive environment.

Behavioral, but not physiological, adaptation to repeated separation in mother and infant primates.

Mother and infant squirrel monkeys were subjected to a series of brief separations in order to evaluate how behavioral and physiological responses change following multiple exposures to stress. Beginning when the infants reached three months of age, their behavioral and hormonal responses were assessed during six 1-hr separations; and additional five dyads served as controls for the effect of repeated disturbance. The separated infants showed a marked and progressive decrease in distress calling across time, but no change was observed in the high levels of agitated activity or the plasma cortisol response to separation. This finding questions the traditional use of distress vocalizations as a measure of stress and indicates that certain types of behavior can change independently of physiological arousal responses, which may continue to occur even after repeated exposures to stress.

Adrenal responses to reinforcement and extinction: role of expectancy versus instrumental responding.

Plasma corticosterone levels were evaluated during operant conditioning in order to determine the effect of reinforced and nonreinforced responding (extinction) on adrenal activation. The influence of instrumental responding was assessed by comparing trained rats with yoked subjects that received a matched reward schedule in the absence of an operant task. Reinforcement sessions resulted in a significant decrease in adrenal secretion at 20 min, but not at 5 min, whereas extinction caused a rapid increase in corticosterone levels at 5 min and an even greater elevation by 20 min. Comparison of the operant and yoked subjects showed that this effect of reinforcement and extinction was not dependent on instrumental responding, but rather on the receipt or withdrawal of the expected reward.

Predictability and coping with separation in infant squirrel monkeys.

Twelve infant squirrel monkeys were separated from their mothers once every 48 hr for a total of 20 separations. Half of the infants experienced separations for a fixed interval (FI) of 30 min. The remaining 6 infants experienced separations for a variable interval (VI) which averaged 30 min. During the separation periods, movement and vocalizations were recorded. Blood was sampled for later cortisol assay after the 1st, 5th, 10th, 15th, and final separation. The FI group showed increased levels of movement and vocalization over repeated separations when compared with the VI group. Cortisol showed significant increases over basal levels and remained elevated throughout the entire period in both groups. These findings suggest that predictability may be more stressful than unpredictability when organisms cannot control the predicted event.

Frontal brain asymmetry and immune function.

The relation between brain activity and the immune system was evaluated by assessing immune responses in 20 healthy women who manifested extreme differences in the asymmetry of frontal cortex activation. One group showed extreme and stable left frontal activation; the other group showed extreme and stable right frontal activation. As predicted, women with extreme right frontal activation had significantly lower levels of natural killer cell activity (at effector:target cell ratios of 33:1 and 11:1) than did left frontally activated individuals. This difference did not extend to two other immune measures, lymphocyte proliferation and T-cell subsets. However, higher immunoglobulin levels of the M class were observed in the right frontal group. In this study, the immune patterns could not be accounted for by plasma cortisol levels, anxiety- and depression-related symptomatology, or recent health histories. These findings support the hypothesis that there is a specific association between frontal brain asymmetry and certain immune responses.

Prenatal influences on neuroimmune set points in infancy.

Many factors during fetal life and early infancy have been found to affect the development of immune responses in animals. This study investigated whether acute exposure of the fetal monkey to high levels of corticosteroids would also have a lingering effect on the expression of immune responses still manifest postpartum in yearling juveniles. One month prior to parturition, pregnant rhesus monkeys were administered dexamethasone for two days. Lymphocyte proliferative responses to mitogen were then examined in their offspring when they were between 1.0-1.5 years of age. In addition, cell sensitivity to corticosteroid feedback was assessed by testing the ability of a gradation of cortisol doses to inhibit proliferation. Monkeys generated from dexamethasone-treated pregnancies tended to have lower responses to concanavalin A. Further, their cells were less sensitive to in vitro incubation with cortisol, suggesting that elevated adrenal activity in vivo had downregulated hormone receptors on their cells. These findings concur with the view that steroidal hormones in utero can influence the fetal immune system, resulting in prolonged effects on immune responses after birth. The similarity of the dexamethasone condition to the clinical treatment used in obstetrical practice raises a potential concern about the widespread antenatal exposure of premature infants to steroidal drugs.

Psychosocial factors and immune senescence in the aged primate.

With increasing age, old animals and humans show decreases in a number of immune responses indicative of the process of immune senescence. Our studies investigated whether social companionship, as a potentially positive psychological intervention, would increase lymphocyte proliferation and natural killer cell activity in the aged nonhuman primate. Contrary to our initial hypothesis, social stimulation resulted in decreased immune responses in old monkeys. With specific modifications of the housing conditions it was possible to prevent these decreases in immune responses from occurring, but social companionship still failed to enhance immune responses in the aged monkey.