Phase specific suppression of neutrophil function in hibernating Syrian hamster.

Hibernation consists of alternating periods of reduced metabolism (torpor) with brief periods of metabolism similar to summer euthermia (arousal). The function of the innate immune system is reduced during hibernation, of which the underlying mechanisms are incompletely understood. Here, we studied neutrophil functionality during hibernation in Syrian hamsters. The inflammatory response to LPS-induced endotoxemia is inhibited in hibernation, partly mediated by reduced IL-6 production in early arousal. Furthermore, neutrophil pathogen binding, phagocytosis and oxidative burst is profoundly reduced in early arousal. Functionality of both summer and early arousal neutrophils was repressed in plasma from early arousal and mixed plasma from early arousal and summer euthermic, but restored by summer euthermic plasma, signifying that a plasma factor in early arousal inhibits TLR-recognition. Identification of the inhibiting factor may offer a target to modulate neutrophil function with relevance to (auto-)inflammatory diseases.

Expression of Paracrine Effectors in Human Adipose-Derived Mesenchymal Stem Cells Treated With Plasma From Brown Bears (Ursus arctos).

Adipose-derived mesenchymal stem cells (ADSCs) are promising candidates for novel cell therapeutic applications. Hibernating brown bears sustain tissue integrity and function via unknown mechanisms, which might be plasma borne. We hypothesized that plasma from hibernating bears may increase the expression of favorable factors from human ADSCs. In an experimental study, ADSCs from patients with ischemic heart disease were treated with interventional media containing plasma from hibernating and active bears, respectively, and with control medium. Extracted RNA from the ADSCs was sequenced using next generation sequencing. Statistical analyses of differentially expressed genes were performed using fold change analysis, pathway analysis, and gene ontology. As a result, we found that genes associated with inflammation, such as IGF1, PGF, IL11, and TGFA, were downregulated by > 10-fold in ADSCs treated with winter plasma compared with control. Genes important for cardiovascular development, ADM, ANGPTL4, and APOL3, were upregulated in ADSCs when treated with winter plasma compared with summer plasma. ADSCs treated with bear plasma, regardless if it was from hibernating or active bears, showed downregulation of IGF1, PGF, IL11, INHBA, IER3, and HMOX1 compared with control, suggesting reduced cell growth and differentiation. This can be summarized in the conclusion that plasma from hibernating bears suppresses inflammatory genes and activates genes associated with cardiovascular development in human ADSCs. Identifying the involved regulator(s) holds therapeutic potential.

Immunoreactivities of NF-κB, IL-1ß and IL-1R in the skin of Chinese brown frog (Rana dybowskii).

The cytokine interleukin-1 beta (IL-1ß) is an evolutionarily conserved molecule that was originally identified in the immune system. Nuclear factor κB (NF-κB) plays a critical role in the activation of immune cells by upregulating the expression of many cytokines. In this study, we investigated the localization and expression level of IL-1ß, its functional membrane receptor type I (IL-1R1) and NF-κB in the skin of Rana dybowskii during the breeding period and pre-hibernation. Histologically, the skin of Rana dybowskii consists of epidermis and dermis, and four kinds of cells were identified in the epidermis during the breeding period and pre-hibernation, while the dermis was composed of homogenous gel, mucous glands and granular glands. IL-1ß, IL-1R1 and NF-κB were immunolocalized in the epithelial and glandular cells in both periods. Western blotting showed that there was no significant difference in the expression of IL-1ß between the breeding period and pre-hibernation, whereas IL-1R1 and NF-κB were significantly higher in the pre-hibernation compared to the breeding period. These results suggested that IL-1ß and NF-κB may collectively play important roles in the skin immune system of Rana dybowskii during the breeding period and pre-hibernation.

Hibernation induces immune changes in the lung of 13-lined ground squirrels (Ictidomys tridecemlineatus).

During hibernation, significant changes occur in the systemic and intestinal immune populations. We found that the lungs of hibernating 13-lined ground squirrels (Ictidomys tridecemlineatus) also undergo shifts in immune phenotype. Within the population of mononuclear cells, the percentage of T cells increases and the percentage of CD11b/c(+) cells decreases in hibernators. E-selectin, which promotes endothelial attachment, increases during arousal from torpor. Levels of the anti-inflammatory cytokine interleukin (IL)-10 in the lung are lower during hibernation while levels of the pro-inflammatory cytokine, tumor necrosis factor (TNF)-α remain constant. Expression of suppressor of cytokine signaling (SOCS) proteins is also decreased in torpid hibernators. Our data point to a unique immune phenotype in the lung of hibernating ground squirrels in which certain immunosuppressive proteins are downregulated while some potentially inflammatory proteins are maintained or amplified. This indicates that the lung houses an immune population that can potentially respond to antigenic challenge during hibernation.

Immunoreactivity of c-kit receptor protein during the prehibernation period in the oviduct of the Chinese brown frog, Rana chensinensis.

The objective of this study was to investigate immunoreactivity of the c-kit receptor in the oviduct of Rana chensinensis during the prehibernation period. Histological examination of oviducts was performed during the prehibernation period. The sections of oviduct were immunostained by the avidin-biotin-peroxidase complex method using rabbit polyclonal antisera raised against the rat c-kit receptor and PCNA. Total proteins were extracted from oviducal tissues and used for Western blotting analysis. Immunohistochemistry revealed the presence of the c-kit receptor and PCNA in the oviduct tissues during the prehibernation period. Also, positive signals for the c-kit receptor and PCNA by Western blotting were observed in oviduct tissues during the prehibernation period. These results suggested that the c-kit receptor might play a regulatory role in oviducal hypertrophy in the brown frog, Rana chensinensis.

Ground squirrel splenic macrophages bind lipopolysaccharide over a wide range of temperatures at all phases of their annual hibernation cycle.

This study evaluates binding of bacterial lipopolysaccharide (LPS) by splenic macrophages from golden-mantled ground squirrels (Spermophilus lateralis, GMGS), a hibernating mammal, at a variety of in vitro incubation temperatures to determine whether this aspect of immune function is effective at low body temperatures. LPS-binding by ground squirrel macrophages was compared to that of rat splenic macrophages. Macrophages were collected from squirrels at discreet stages in their annual cycle and incubated with fluorescein-labeled LPS (LPS-FITC). The percentage of GMGS that bound LPS-FITC did not change as a function of hibernation season or as a function of incubation temperature. The total amount of LPS-FITC bound per cell was similarly unaffected by season or temperature, however, macrophages from torpid squirrels bound more LPS-FITC than cells from normothermic squirrels. Macrophages of golden-mantled ground squirrels bind LPS at a wide range of temperatures throughout their annual cycle; an ability shared between hibernators and non-hibernators alike.

Production of tumor necrosis factor in cells of hibernating ground squirrels Citellus undulatus during annual cycle.

TNF production has been studied in peritoneal macrophages and splenic T cells of Arctic Yakutian ground squirrel (Citellus Undulatus Pallas) in hibernating and awake animals in winter and in prehibernating autumn as well as in active euthermic spring-summer animals. A high level of TNF production in macrophages of ground squirrel is observed over the active period and during arousals in winter. There are no significant season variations in TNF production in splenic T lymphocytes of ground squirrels. This suggests the major role of activated macrophages in the arousals of hibernating animals. T lymphocyte proliferation in ground squirrels in the active period is higher than in winter, and the most significant seasonal variations are found in T cell mitogenic response, which increases in spring-summer period. Evidence is presented that functional activity of macrophages of squirrel in autumn has much in common with that in winter rather than in spring-summer period.