Alba Rossi-George

Neuronal, Endocrine, and Anorexic Responses to the T-Cell Superantigen Staphylococcal Enterotoxin A: Dependence on Tumor Necrosis Factor-α

Staphylococcal enterotoxin A (SEA) is a microbial superantigen that activates T-lymphocytes and induces production of various cytokines, including tumor necrosis factor-α (TNFα). Previously, it was shown that SEA activates the hypothalamic-pituitary-adrenal axis and augments gustatory neophobic behaviors. In the present study, it was hypothesized that these effects involve neuronal activation in forebrain regions mediating fear and/or anxiety and are dependent on the production of TNFα. Male C57BL/6J mice were given intraperitoneal injections of 10 μg of SEA and 5 μg of lipopolysaccharide (LPS) or saline and perfused 2 h later for histochemical determination of brain c-Fos immunoreactivity (IR). The results showed increased c-Fos IR in the paraventricular nucleus, arcuate nucleus, central nucleus of the amygdala, bed nucleus of the stria terminalis, and lateral septum. Challenge of TNF-/- mice with SEA did not produce a significant increase in brain c-Fos IR, although c-Fos was increased after exposure to a psychogenic stressor (i.e., open field). In additional experiments, the elevated corticosterone response to SEA was abrogated in TNF-/- mice and was shown to be corticotropin-releasing hormone dependent. Finally, the augmented reduction in novel food intake after SEA challenge was attenuated in TNF-/- mice as well as in wild-type mice administered antibody to TNFα. In conclusion, challenge with SEA recruits brain regions mediating stress and anxiety responses, an effect that requires endogenous TNFα. Whether this is indicative of all T-cell superantigens remains to be determined, although it stands in contrast to other models of neuroimmunomodulation (e.g., LPS) that involve multiple cytokine influences.

Effects of bacterial superantigens on behavior of mice in the elevated plus maze and light-dark box

Bacterial superantigens, such as staphylococcal enteroxins A and B (SEA/SEB) stimulate T cells to produce high levels of cytokines in blood. Previously it had been shown that these toxins were capable of stimulating increased neuroendocrine activity and enhanced behavioral reactivity to novel gustatory and non-gustatory stimuli. Therefore, it was suggested that these superantigens may promote anxiety-like behavior. In the current set of experiments, BALB/cByJ and C57BL/6J male mice were challenged with either SEB (50 microg) or SEA (5 or 10 microg) and tested for behavior in the elevated plus maze (EPM). Results suggested an absence of increased anxiety-like behavior, with exploration of the open arms being enhanced by SEA or SEB treatment. In another test of anxiety, the light-dark box, SEB challenge of BALB/cByJ mice 90 min prior to testing, did not alter exit latency, activity nor time spent in the dark. However, in a second experiment, it was found that if animals were first tested for consumption, followed by testing in the light-dark box, SEB challenged animals displayed increased exit latency and reduced exploration. These studies suggest that in standard tests of rodent anxiety-like behavior, evidence for the induction of anxiety-like processes subsequent to challenge with SEA or SEB is not patently discernable. However, neurobiological events induced by immunological challenge might synergize with reactivity to psychogenic and/or gustatory stimuli, thereby resulting in increased anxiety-like behavior that could be unmasked by standard behavioral tests such as the light-dark box or EPM.

Retroactive interference in 3-month-old infants

Three experiments assessed the effect of the similarity and timing of interpolated information on retroactive interference in human infants. After 3-month-olds learned to move a mobile (the cue) in a distinctive context by kicking, they were exposed to novel stimuli and were tested 24 hr later for recognition of their training stimuli. In Experiment 1, the interpolated cue, context, or both were novel. Retroactive interference occurred unless the interpolated and training stimuli shared no components. In Experiments 2 and 3, the timing of exposure to the interpolated context or cue, respectively, varied. A novel context impaired recognition after exposure delays up to 2 hr, whereas a novel cue impaired recognition after exposure delays up to 40 min. The finding that the cue is less vulnerable to retroactive interference than the context suggests that it is processed more rapidly. These experiments reveal that retroactive interference in infants depends on both the similarity and timing of the interpolated information.

Stressor‐induced modulation of immune function: a review of acute, chronic effects in animals

The present paper reviews recent studies on the effects of stress on immune function in laboratory animals. The emphasis is on those studies where a simultaneous comparison of acute and chronic stress regimens was determined, although additional relevant studies are also reviewed. The effects of stress on basic measurements of cellular and humoral immune measures are discussed, including the growing number of studies that have reported alterations in macrophage functions. The latter are key elements in the innate immune response, and like measurements of T cell function and antibody production, are inhibited and enhanced by stressor exposure. This review does not focus on the mechanisms by which stress alters immune function, there being little to add conceptually in terms of what was reported previously (see Kusnecov AW, Rabin BS, Int Arch Allergy Immunol 1994;105:107–121.). However, a question is raised in the conclusion as to how stressor effects on immune function should be interpreted, for it is clear that immunological processes in and of themselves elicit central nervous system responses that neurochemically and endocrinologically do not differ from those produced in response to psychological stressors. Therefore, at least in the short term stressor-induced immune changes may not necessarily reflect maladaptive adjustments, although, as demonstrated by some studies reviewed in this paper, they may pose a serious risk to health should stressor exposure be persistent and uncontrolled.

Copper modulates the phenotypic response of activated BV2 microglia through the release of nitric oxide

Microglia are resident immune cells of the central nervous system. Their persistent activation in neurodegenerative diseases, traditionally attributed to neuronal dysfunction, may be due to a microglial failure to modulate the release of cytotoxic mediators such as nitric oxide (NO). The persistent activation of microglia with the subsequent release of NO vis-á-vis the accumulation of redox transition metals such as copper (Cu) in neurodegenerative diseases, prompted the hypothesis that copper would alter NO signaling by changing the redox environment of the cell and that, by altering the fate of NO, microglia would adopt a different phenotype. We have used the microglial cell model, BV2, to examine the effects of Cu(I) on NO production and activation as they have been shown to be phenotypically plastic. Our results show that cell viability is not affected by Cu(I) in BV2 microglia and that it has no effect on iNOS mRNA, protein expression and nitrite release. However, when LPS is added to Cu(I)-treated medium, nitrite release is abrogated while iNOS expression is not significantly altered. This effect is Cu(I)-specific and it is not observed with other non-redox metals, suggesting that Cu(I) modulates NO reactivity. Immunofluorescence analysis shows that the M1 (inflammatory) phenotype of BV2 microglia observed in response to LPS, is shifted to an M2 (adaptive) phenotype when Cu(I) is administered in combination with LPS. This same shift is not observed when iNOS function is inhibited by 1400W. In the present study we show that Cu(I) modulates the release of NO to the media, without altering iNOS expression, and produces phenotypic changes in BV2 microglia.

Dissipation of retroactive interference in human infants

In 3 experiments with 85 human 3-month-olds, the authors asked whether retroactive interference with their memory of the original training stimulus is temporary or permanent. Infants learned to move a mobile by kicking and then were exposed to a different mobile (Experiment 1) or context (Experiment 2) immediately or 3 days afterward (Experiment 3). They were tested after increasing delays with the original stimulus, the exposed stimulus, or a completely novel stimulus. Retroactive interference was temporary and unrelated to the exposure delay. The data are consistent with a retrieval-based account of interference. Memory updating (i.e., responding to the interfering stimulus) was coincident with retroactive interference, suggesting that retroactive interference is an adaptive mechanism that facilitates memory updating within a narrow time window.

Potentiation of Interleukin-1β Adjuvant Effects on the Humoral Immune Response to Antigen in Adrenalectomized Mice

Objectives: Activation of the hypothalamic-pituitary-adrenal (HPA) axis is a primary effect of interleukin-1 (IL-1), with elevated glucocorticoids considered a mechanism for negative feedback immunoregulation. However, there is little direct evidence of such a functional relationship between IL-1-mediated immunoregulation and neuroendocrine influences elicited by IL-1. Therefore, the goal of this study was to examine whether the known adjuvant effects of IL-1 are altered in the absence of neuroendocrine feedback due to adrenalectomy. Methods: Male BALB/c mice subjected to adrenalectomy (ADX) or sham surgery were administered with saline or recombinant human IL-1β (rhIL-1β) and at the same time immunized with 100 µg ovalbumin (OVA). In vivo and in vitro measures of antigen-specific IgG antibody production, IL-6 production and spleen cell proliferation were taken 6 and 12 days later. Results: It was demonstrated that administration of rhIL-1β at a dose that activates the HPA axis resulted in a significant augmentation of serum anti- OVA IgG antibody levels. Interestingly, this augmentation was potentiated in ADX animals. In addition, the in vitro spleen cell memory IgG antibody response to OVA was significantly augmented in rhIL-1β-treated animals, and again, further potentiated in ADX animals. Interestingly, while hrIL-1β treatment augmented antigen-stimulated IL-6 production – suggesting an effect of IL-1 on antigen-specific T helper 2 cell memory formation – potentiation was not evident in ADX animals. Conclusions: These results are consistent with the concept of HPA axis-mediated neuroendocrine feedback on excessive immune responsiveness due to IL-1. Such feedback may prevent disturbances to the self-limiting functions of the immune system, which are important to the prevention of autoimmune diseases, some of which involve elevated IL-1 production.

Immunofluorescent Detection of S-nitrosoproteins in Cell Culture

The role of S-nitrosylation in cellular signaling has been clearly demonstrated. There a number of mechanisms whereby this post-translational modification can occur and the number of protein targets continue to expand. The need to be able to monitor when this important signaling process occurs within cells is increasingly important. Previously we have identified immunohistochemistry approaches effective for monitoring S-nitrosylation within fixed tissue. Within this paper we show how these techniques can be adapted to use in a cell culture system using immunofluorescence. We have used this protocol to detect S-nitrosoprotein formation within LPS stimulated microglial cells using both transformed and primary cultured cells.

Nitric Oxide Biochemistry: Pathophysiology of Nitric Oxide-Mediated Protein Modifications

Nitric oxide (NO) is a pluripotent signaling molecule, which has been proposed to be critically important in both physiological and pathological processes of the brain. The wide-ranging functionality of NO is in opposition to its relatively simple chemical structure. In this chapter we attempt to summarize the functional involvement of NO within the neurological system and then discuss how such complex signaling may be achieved via the differential post-translational modification of protein targets. It is our contention that the redox properties of NO allow this molecule to modify proteins in a variety of ways with physiological or pathological consequences. In this way the effects of NO production are dependent on the quantity produced, the redox environment in which it is synthesized, and the presence of reactive targets. A summary of known post-translational modifications is given as well as the functional consequences of their formation.