IL-5 but not interferon-gamma (IFN-γ) inhibits eosinophil apoptosis by up-regulation of bcl-2 expression

In order to determine regulatory mechanisms of eosinophil apoptosis, we examined the effect of recombinant IL-5 and interferon-gamma (IFN-γ) on eosinophil apoptosis and bcl-2 expression. rhIL-5 (2.5 ng/ml) significantly inhibited eosinophil apoptosis in 96 h in vitro culture compared with medium only-cultured eosinophils (89.4 ± 3.6% versus 31.3 ± 12.2% (mean ± s.d.); n = 7, P < 0.05). Further, rhIL-5 significantly increased bcl-2 protein and mRNA expression on cultured eosinophils. A phosphorothioate antisense oligonucleotide targeted at the ATG translation initiation codon of bcl-2 (10⁻⁵ m) could significantly block the supportive effect of rhIL-5 (0.25 ng/ml) for eosinophil survival compared with sense cDNA of bcl-2 on 96 h culture (inhibition rate 28.01 ± 4.56% versus 0.07 ± 1.73%; n = 4, P < 0.05). In contrast, rhIFN-γ (100 U/ml) significantly inhibited eosinophil apoptosis on 96 h in vitro culture (72.7 ± 10.5%; n = 7, P < 0.05), but did not significantly up-regulate bcl-2 protein and mRNA. These results indicate that IL-5 has inhibitory effects on eosinophil apoptosis by regulation of bcl-2 expression.     

Analysis Tissue Expression of IFN-γin IL-12 and／or IL-18 Gene Ablated Naieve Mice

Interleukin 12 (IL-12) and/or interleukin 18 (IL-18) gene ablated mice were applied for the investigation of the tissue expression of interferon gamma (IFN-gamma). For IL-12(-/-), IL-18(-/-), IL-12(-/-)/18(-/-) and wt mice, reproductive performance were recorded and IFN-gamma concentrations in heart, lung, liver, spleen, kidney and serum were quantified by ELISA. There were no significant differences of IFN-gamma in heart, lung and kidney between 4 strains although control group was higher. It was observed that for IL-12(-/-) mice, compared with other 3 groups, IFN-gamma in liver and spleen were decreased (p < 0.05) and reproductive performance appeared to be impaired. Serum IFN-gamma level of IL-12(-/-)/18(-/-) mice was significantly higher (p < 0.05). It was showed that IFN-gamma productions under the normal condition were independent upon IL-12 and IL-18, its expressions in various tissues were different, and optimal IFN-gamma is necessary for the normal growth and development of mammals. This study is helpful for clinical cytokines therapy.     

Effects of combination of Cryptococcus gattii and IFN-γ, IL-4 or IL-27 on human bronchial epithelial cells

BackgroundAirway epithelial cells are crucial for the establishment of cryptococcosis. In experimental cryptococcosis, the Th2 immune response is associated with host susceptibility, while Th1 cells are associated with protection. The absence of IL-27 receptor alpha in mice favor the increase Cryptococcus neoformans burden in the lung. Here, we evaluated the effects of the combination of IL-4, IFN-γ or IL-27 with C. gattii on human bronchial epithelial cells (BEAS-2B).MethodsBEAS-2B were stimulated with IL-4, IFN-γ or IL-27 (100 ng/mL) and/or live yeast forms of C. gattii (multiplicities of infection (MOI) of 1–100) and vice-versa, as well as with heat-killed cells of C. gattii for 24 h.ResultsNone of the C. gattii MOIs had cytotoxic effects on BEAS-2B when compared to control. The cells stimulated by cytokines (IL-4, IFN-γ or IL-27) followed by live yeast forms of C. gattii (MOI of 100) infection and vice-versa demonstrated a reduction in IL-6, IL-8 and/or CCL2 production and activation of STAT6 (induced by IL-4) and STAT1 (induced by IL-27 or IFN-γ) when compared to cells stimulated with C. gattii, IL-4, IFN-γ or IL-27. In the combination of cytokines and heat-killed cells of C. gattii, no inhibition of these inflammatory parameters was observed. The growth of C. gattii was increased while the phagocytosis of live yeast forms of C. gattii in the BEAS-2B were reduced in the presence of IL-4, IFN-γ or IL-27.ConclusionThe association of live yeast forms, but not heat-killed yeast forms, of C. gattii with IL-4, IFN-γ or IL-27 induced an anti-inflammatory effect.     

MAPKs ERK and p38, but not JNK phosphorylation, modulate IL-6 and TNF-α secretion following OK-432 in vitro stimulation of purified human monocytes

Interaction between the immune system and cancer allows for the use of biological response modifiers, e.g. OK-432, in cancer therapy. OK-432, penicillin-killed Streptococcus pyogenes, is used in treating carcinomas, but also lymphangiomas. We have studied the role of monocytes (MOs) in the immune response to OK-432 by examining IL-6 and tumour necrosis factor (TNF)-α secretion after in vitro MO stimulation with OK-432, to some extent in comparison with lipoteichoic acid (LTA) and lipopolysaccharide (LPS). LTA stimulation of whole blood gave IL-6 but not TNF-α secretion, as previously shown with OK-432 stimulation, whereas both cytokines were secreted following LPS stimulation. Addition of the MAPK kinase (MAPKK) MEK inhibitor U0126 inhibited IL-6/TNF-α secretion in a dose-dependent manner. Flow cytometry and to some extent Western blot (Wb) analyses showed that MAPK ERK, located downstream of MEK1/2, is predominantly phosphorylated at isolation from peripheral blood. Addition of the p38 MAP kinase inhibitor SB202190 decreased MO IL-6/TNF-α production upon OK-432 stimulation in a dose-dependent manner. Addition of the MAPK JNK inhibitor SP600125 did not systematically change the MO IL-6/TNF-α OK-432 response. Flow cytometry showed that when stimulating the MOs before isolation from blood, LPS yielded ERK phosphorylation and LPS/LTA p38 phosphorylation, whereas OK-432 had no effects on phosphorylation levels. In conclusion, we have shown that OK-432 resembles TLR2 more than TLR4 stimulation of MOs and depends on MAPKK MEK and MAPK p38, but not on JNK phosphorylation. The MEK and p38 MO OK-432 stimulation dependence is possibly related to the differentiation of cells of the MO lineage.     

Induction of metastatic cancer stem cells from the NK/LAK-resistant floating, but not adherent, subset of the UP-LN1 carcinoma cell line by IFN-γ

As an advanced status of cancer stem cells (CSCs), metastatic CSCs (mCSCs) have been proposed to be the essential seeds that initiate tumor metastasis. However, the biology of mCSCs is poorly understood. In this study, we used a lymph node (LN) metastatic CEA-producing carcinoma cell line, UP-LN1, characterized by the persistent appearance of adherent (A) and floating (F) cells in culture, to determine the distribution of CSCs and mechanisms for the induction of mCSCs. F and A cells displayed distinct phenotypes, CD44(high)/CD24(low) and CD44(low)/CD24(high), respectively. The CSC-rich nature of F cells was typified by stronger expression of multiple drug resistance genes and a 7.8-fold higher frequency of tumor-initiating cells in NOD/SCID mice when compared with A cells. F cells showed a greater depression in HLA class I expression and an extreme resistance to NK/LAK-mediated cytolysis. Moreover, the NK/LAK-resistant F cells were highly susceptible to IFN-γ-mediated induction of surface CXCR4, with concomitant downregulation of cytoplasmic CXCL12 expression, whereas these two parameters remained essentially unchanged in NK/LAK-sensitive A cells. Following the induction of surface CXCR4, enhanced migratory/invasive potential of F cells was demonstrated by in vitro assays. Confocal immunofluorescence microscopy showed the two distinct phenotypes of F and A cells could be correspondingly identified in monodispersed and compact tumor cell areas within the patient's LN tumor lesion. In response to IFN-γ or activated NK/LAK cells, the CXCR4(+) mCSCs could be only induced from the CSCs, which were harbored in the highly tumorigenic CD44(high)/CD24(low) F subset. Our results revealed the complexity and heterogeneity of the CSC of this cell line/tumor and the differential immunomodulatory roles of F and A cells. A better understanding of the interactions among different classes of CSCs and their niches may assist us in eradicating the CSCs/mCSCs through targeted immunotherapy, chemotherapy, or both.     

Dendritic cells treated with HPV16mE7 in a three-dimensional model promote the secretion of IL-12p70 and IFN-γ

Although the human papillomavirus (HPV) DNA therapeutic vaccine represents a promising approach to the prevention and treatment of cervical cancer, the mechanism of the HPV DNA vaccine is poorly understood. Moreover, current strategies have met with only limited success in preclinical and dendritic cell-based (DC-based) clinical research. In addition, two-dimensional (2-D) DC monolayers poorly mimic the physiology function in vivo. We used a three-dimensional (3-D) DC culture model in vitro to explore the immune mechanism of the HPV DNA vaccine. DCs were generated from peripheral blood monocytes with interleukin-4 (IL-4) and granulocyte-macrophage colony-stimulating factor (GM-CSF). The cells, growing in 3-D collagen gel, were treated with pcDNA3.1-HPV16mE7 in vitro for 48 h. Compared to DCs treated with E7 in a 2-D culture model, the expression of co-stimulatory molecules CD80 and CD40 were significantly increased in the 3-D model (p < 0.05), and a remarkable increase of IL-12 p70 was observed. However, we did not detect any obvious change in IL-10 in 3-D culture. In addition, we found that IFN-γ expression increased when HPV16mE7-DC cells were co-cultured with T-cells for 96 h in the 3-D model, and HPV16mE7-DCs stimulated the proliferation of T lymphocytes more efficiently in the 3-D model than in the 2-D model (p < 0.05). These results suggest that DCs in 3-D culture model have a notable effect on the enhancement of the HPV16 DNA vaccine's immune reaction and indicate that the DC-based 3-D model is a novel approach to study the HPV vaccine.     

Analysis Tissue Expression of IFN-γin IL-12 and/or IL-18 Gene Ablated Na(?)ve Mice

Interleukin 12(IL-12)and/or interleukin 18(IL-18)gene ablated mice were applied for the investigation of thetissue expression of interferon γ(IFN-γ).For IL-12~(-/-),IL-18~(-/-),IL-12~(-/-)/18~(-/-) and wt mice,reproductive performancewere recorded and IFN-γ concentrations in heart,lung,liver,spleen,kidney and serum were quantified by ELISA.There were no significant differences of IFN-γ in heart,lung and kidney between 4 strains although control groupwas higher.It was observed that for IL-12~(-/-) mice,compared with other 3 groups,IFN-γ in liver and spleen weredecreased(p<0.05)and reproductive performance appeared to be impaired.Serum IFN-γ level of IL-12~(-/-)/18~(-/-)mice was significantly higher(p<0.05).It was showed that IFN-γ productions under the normal condition wereindependent upon IL-12 and IL-18,its expressions in various tissues were different,and optimal IFN-γ is necessaryfor the normal growth and development of mammals.This study is helpful for clinical cytokines therapy.Cellular& Molecular Immunology.2005;2(1):68-72.     

Modulation of phagocytic and oxidative burst activities of bovine neutrophils by human recombinant TNF-α, IL-1-α, IFN-γ, G-CSF, GM-CSF

In vitro experiments were conducted to determine the effects of recombinant cytokines on phagocytic and oxidative burst activities of bovine neutrophils. Neutrophils were isolated (purity >91%, viability >97%) from EDTA-anticoagulated blood from healthy Holstein-Friesian heifers. Aliquots of neutrophils (10 × 10⁶ cells/ml) were incubated for 1 h at 37 °C with equal volumes of recombinant human cytokines, namely, tumour necrosis factor-alpha (rhTNF-, 0.5–1000 ng/ml), interleukin-1-alpha (rhIL-1-, 0.001–10 ng/ml), interferon-gamma (rhIFN-, 0.01–100 ng/ml), granulocyte colony-stimulating factor (rhG-CSF, 25 ng/ml), and granulocyte-macrophage colony-stimulating factor (rhGM-CSF, 10 ng/ml). Then, the percentage phagocytosis and average number of intracellular bacteria per cell were evaluated by flow cytometry and/or fluorescent microscopy using FITC-labelled opsonised bacteria (Escherichia coli 0111:B4). Unlabelled opsonised bacteria and dichlorofluorescin diacetate were used to evaluate H₂O₂ production, a measure of oxidative burst, by flow cytometry. The results showed that all five cytokines significantly (p <0.05) increased percentage phagocytosis (52.4–86.1%), number of intracellular bacteria per cell (24.9–47.9%), and H₂O₂ production (31.3–58.2%) when compared to untreated neutrophils. A gradual increase in mean channel fluorescence but not in percentage phagocytosis was consistently seen with increasing concentrations of rhTNF-, rhIL-1-, and rhIFN-, thereby indicating a concentration-dependent stimulation of phagocytic capacity by these three cytokines.     

IFN-β, IFN-γ, and TNF-α decrease erythrophagocytosis by human monocytes independent of SIRP-α or SHP-1 expression

Background: Many cases of autoimmune hemolytic anemia have been reported after viral infection. Phagocyte activation and accompanying erythrophagocytosis are thought to result from proinflammatory cytokines released during viral infection. SIRP-α (signal regulatory protein-α), a receptor expressed on phagocytes, inhibits phagocytosis when bound to CD47 on the erythrocyte membrane. Ligation with CD47 results in SHP-1 recruitment to SIRP-α and dephosphorylation of specific downstream substrates involved in phagocytosis. SIRP-α ligation by CD47 may be inhibited by proinflammatory cytokines.

Objectives: The aim of this work was to evaluate the effect of IFN-β, IFN-γ, and TNF-α on erythrophagocytosis and assess the effect on expression of SIRP-α and SHP-1 in human monocytes.

Materials and methods: Monocytes were cultured ex vivo with IFN-β or IFN-γ/TNF-α. Erythrophagocytosis was determined by flow cytometry. SIRP-α and SHP-1 gene expression was determined by real time-PCR, while SIRP-α and SHP-1 protein expression was determined by western blot.

Results: Erythrophagocytosis by monocytes significantly decreased after treatment with either IFN-β or IFN-γ/TNF-α. Monocytes cultured with IFN-γ/TNF-α showed increased SIRP-α gene and protein expression and SHP-1 gene expression. Monocytes cultured with IFN-β did not show any alteration in SIRP-α or SHP-1 expression.

Conclusion: We conclude that IFN-β and IFN-γ/TNF-α decrease erythrophagocytosis by human monocytes in vitro, and this effect does not apparently require an increase in SIRP-α or SHP-1 expression.     

TSLP induced by estrogen stimulates secretion of MCP-1 and IL-8 and growth of human endometrial stromal cells through JNK and NF-κB signal pathways

It has reported that human endometrial stromal cells (ESCs) express thymic stromal lymphopoietin (TSLP), and TSLP concentrations in the serum and peritoneal fluid were higher in women with endometriosis. Endometriosis is an estrogen-dependent disease. The present study aimed to elucidate whether and how estrogen regulates the growth of ESCs through TSLP. The ESCs behaviors in vitro were verified by SRB assay and Ki67 level detection, respectively. In addition, the effects of estrogen on TSLP and TSLP on the correspondent functional molecules were investigated by ELISA and flow cytometry. Here we found that estrogen stimulated the secretion of TSLP in a dosage-dependent manner. Recombinant human TSLP stimulates the secretion of MCP-1 and IL-8, and markedly promotes the viability and proliferation relative gene Ki-67 expression of ESCs. These effects could be abolished by the inhibitor for JNK or NF-κB signal, respectively. Moreover, not only anti-TSLP neutralizing antibody, but also blocking JNK or NF-κB signal by inhibitor abrogated the stimulatory role in the production of MCP-1 and IL-8, and the growth of ESCs induced by estrogen. Our current study has demonstrated that TSLP is involved in the regulation of estrogen on the secretion of MCP-1 and IL-8, and the growth of ESCs through JNK and NF-κB signal pathways, which suggests that the abnormal high expression of TSLP induced by estrogen may play an important role in ESCs growth and finally contribute to the origin and development of endometriosis.     

Inhibition of tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) secretion but not IL-6 from activated human peripheral blood monocytes by a new synthetic demethylpodophyllotoxin derivative

A newly synthesized demethylpodophyllotoxin derivative, 4-O-butanoyl-4-demethylpodophyllotoxin (BDPT) or BN58705, has recently been shown to exert a potent cytotoxic activityin vitro against a variety of drug-resistant human tumor cell lines. The effect of this agent on effector cells of the immune system, however, has not been examined. The present study investigated the effect of BDPT on the response of activated human peripheral blood derived monocytes (PBM) to secrete cytokines. Activation of PBM overnight with LPS, IFN-, or PMA resulted in secretion into the supernatant of TNF-, IL-1, IL-6, and IL-8 as assessed by ELISA. The addition of BDPT to the stimulated cultures resulted in significant inhibition of TNF- and IL-1 secretion, whereas the secretion of IL-6 and IL-8 was not affected. The selective inhibition of TNF- and IL-1 secretion by BDPT-treated PBM was observed with all three stimuli tested. The inhibitory effect mediated by BDPT was concentration dependent and was optimal at 6–20 M. Time kinetic analysis indicated that the inhibition of secretion was rapid and detected as soon as 2 hr following stimulation of the PBM and lasted for as long as 24 hr. A comparison was made between BDPT and pentoxyfilline, a xanthine-derived phosphodisterase inhibitor that was reported to inhibit TNF- and IL-1 secretion by PBM. Both BDPT and PTX showed similar time kinetics and patterns of inhibition. However, the concentration used by BDPT to achieve optimal inhibition of secretion was 10- to 20-fold less than that needed by PTX. The selective inhibition of TNF- secretion by BDPT and PTX was corroborated by inhibition of TNF- mRNA but not IL-6 mRNA as assessed by RT-PCR analysis. These studies demonstrate that the antitumor cytotoxic compound BDPT is also an immunomodulatory agent that can inhibit selectively TNF- and IL-1 secretion by PBM. Further, the low toxicity and low concentrations of BDPT needed for optimal inhibition suggest that BDPT may have potential in its therapeutic application in diseases that are mediated by TNF- and IL-1 like septic shock, inflammatory responses, and infections.     

Dopamine, norepinephrine, and the management of sensorimotor bindings: individual differences in updating of stimulus–response episodes are predicted by DAT1, but not DBH5′-ins/del

Evidence suggests that the flexibility of managing (creating and updating) stimulus–response bindings is driven by the dopaminergic system. Given that striatal dopamine (DA) plays a crucial role in the updating of working memory, the present study tested whether individual differences in the efficiency of updating stimulus–response episodes (event files) are predicted by differences in genetic predisposition related to the efficiency of the striatal dopaminergic pathway. In view of contrasting claims that stimulus–response binding is related to norepinephrine, we also considered genetic predispositions regarding noradrenergic pathways. In a sample of 100 healthy adults, we studied whether the degree to which stimulus–response bindings affect ongoing performance is predicted by polymorphisms of the dopamine transporter gene (DAT1, associated with striatal DA levels) and DBH5′-ins/del (strongly correlated with dopamine beta-hydroxylase, the enzyme catalyzing the dopamine-norepinephrine conversion). The performance of 9-repeat carriers of the DAT1 gene was more affected by stimulus–response bindings than the performance of 10/10 homozygotes was, while DBH5′-ins/del polymorphism was not related to performance. This outcome pattern suggests a crucial role of the nigrostriatal dopaminergic pathway in the flexible management of stimulus–response episodes, whereas norepinephrine does not seem to play a role.     

IL-18/IL-15/IL-12 synergy induces elevated and prolonged IFN-γ production by ex vivo expanded NK cells which is not due to enhanced STAT4 activation

The synergistic effect of IL-18/IL-15/IL-12 stimulation potently activates NK cells, inducing high levels of IFN-γ production. As a result of this potent stimulatory effect, NK cell pre-activation with IL-18/IL-15/IL-12 is being developed as a cancer immunotherapy. Ex vivo expansion of NK cells enables the efficient generation of large numbers of NK cells for wide-scale and repeated therapeutic use, and is thus an important source of NK cells for clinical application. However, the effects of IL-18/IL-15/IL-12 stimulation on ex vivo expanded NK cells have not yet been assessed. Thus, the present study assessed the effects of IL-18/IL-15/IL-12 stimulation on NK cells expanded ex vivo using K562-based artificial antigen presenting cells expressing membrane-bound IL-21. We report that ex vivo expanded NK cells stimulated with IL-18/IL-15/IL-12 produce high levels of IFN-γ and TNFα, have potent cytotoxicity, and maintain prolonged IFN-γ production following removal of stimulation. IL-18/IL-15/IL-12 stimulation induces a phenotypically unique IFN-γ-producing population with reduced CD16 expression and greater CD25 expression as compared to stimulated IFN-γ- NK cells and unstimulated NK cells. We elucidate that the mechanism of synergy for induction and maintenance of IFN-γ production is not due to a further enhancement of STAT4 activation compared to stimulation with IL-12 alone. Furthermore, we demonstrate that the synergistic increase in IFN-γ is not solely under translational regulation, as elevated levels of IFN-γ mRNA contribute to the synergistic increase in IFN-γ. Overall, this study characterizes the response of ex vivo expanded NK cells to IL-18/IL-15/IL-12 stimulation and supports the use of ex vivo expanded NK cells as a feasible and efficient source of IL-18/IL-15/IL-12 pre-activated NK cells for adoptive transfer in cancer immunotherapies.     

Modulation of the Functions of Dengue Virus-Specific Human CD8+ Cytotoxic T Cell Clone by IL-2, IL-7 and IFNγ

Lymphokines play an important role in immune responses to viruses by modulating functions of T lymphocytes. In the present study, we examined the effects of interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-7 (IL-7), and interferon gamma (IFNγ) on proliferation, cytotoxic activity and lymphokine production of a dengue virus-specific CD8⁺ human cytotoxic T lymphocyte (CTL) clone. IL-2 and IL-7 induced proliferation of the CD8+ CTL clone in the presence or absence of specific antigen, while IFNγ suppressed proliferation of the clone. IL-7 and IFNγ augmented dengue virus-specific cytotoxic activity without inducing non-specific cytotoxic activity, and IL-2 induced non-specific cytotoxic activity. IL-2 induced IFNγ production by the CD8+ CTL clone. IL-4 and IL-6 did not modulate the functions of the CD8+ CTL clone in these experimental conditions.

These results suggest that functions of dengue virus-specific CD8+ CTL are modulated by IL-2, IL-7 and IFNγ, and that IL-7 is a lymphokine useful to induce growth and to maintain specific cytotoxic activity of CD8+ CTL clones in vitro.     

Involvement of glutamate NMDA and AMPA receptors, glial cells and IL-1β in the spinal hyperalgesia evoked by the chemokine CCL2 in mice

We study here the involvement of excitatory amino acid receptors, glial cell activation and IL-1β release in the spinal hyperalgesia evoked by the chemokine CCL2 (MCP-1). Three hours after the intrathecal administration of CCL2 (1-100 ng), mice exhibit dose-dependent thermal hyperalgesia, that was inhibited by the coadministration of the antagonist of chemokine receptor type 2 (CCR2) RS504393 (0.3-3 μg). To assess the involvement of excitatory amino acid receptor sensitisation, CCL2 was coadministered with CPP (0.3-3 ng) and NBQX (25-250 ng), antagonists of NMDA and AMPA receptors, respectively. Both drugs blocked CCL2-evoked hyperalgesia, strongly suggesting that CCL2 evokes in vivo NMDA and AMPA receptor sensitisation, as previously described in electrophysiological studies. Furthermore, this rapid induction of CCL2-mediated hyperalgesia was blocked by the previous acute administration of the microglial inhibitor minocyclin (4.9 μg) or the astroglial inhibitor l-aminoadipate (1.6 μg). Since IL-1β can be released by activated glial cells we tested whether this cytokine could be underlying the spinal sensitisation induced by CCL2. The administration of the type I IL-1 receptor antagonist, IL-1ra (3-30 μg), partially prevented CCL2-evoked hyperalgesia. Finally, to elucidate if IL-1β could produce NMDA and AMPA receptor sensitisation by itself, we performed experiments in which this cytokine was i.t. administered. Thermal hyperalgesia induced by IL-1β (30 pg) was completely prevented by the coadministration of CPP (3 ng) but unaffected by NBQX (250 ng). Globally, our results suggest that spinal CCL2 induces thermal hyperalgesia by sensitising NMDA and AMPA receptors in a process that involves glial activation and IL-1β release.     

To eat or not to eat? Kinematics and muscle activity of reach-to-grasp movements are influenced by the action goal, but observers do not detect these differences

Recent evidence suggests that the mirror neuron system responds to the goals of actions, even when the end of the movement is hidden from view. To investigate whether this predictive ability might be based on the detection of early differences between actions with different outcomes, we used electromyography (EMG) and motion tracking to assess whether two actions with different goals (grasp to eat and grasp to place) differed from each other in their initial reaching phases. In a second experiment, we then tested whether observers could detect early differences and predict the outcome of these movements, based on seeing only part of the actions. Experiment 1 revealed early kinematic differences between the two movements, with grasp-to-eat movements characterised by an earlier peak acceleration, and different grasp position, compared to grasp-to-place movements. There were also significant differences in forearm muscle activity in the reaching phase of the two actions. The behavioural data arising from Experiments 2a and 2b indicated that observers are not able to predict whether an object is going to be brought to the mouth or placed until after the grasp has been completed. This suggests that the early kinematic differences are either not visible to observers, or that they are not used to predict the end-goals of actions. These data are discussed in the context of the mirror neuron system.     

Prostaglandin D2 and J2-series (PGJ2, Δ12-PGJ2) prostaglandins stimulate IL-6 and MCP-1, but inhibit leptin, expression and secretion by 3T3-L1 adipocytes

Prostaglandin D₂ and its derivatives PGJ₂ and Δ¹²-PGJ₂ strongly stimulate the synthesis and secretion by white adipocytes of the neurotrophin NGF. Here we have explored whether PGD₂ and the J₂-series prostaglandins have pervasive effects on adipokine production. The influence of these prostaglandins on the production of the adipocyte hormones leptin and adiponectin, and the inflammatory factors IL-6 and monocyte chemoattractant protein 1 (MCP-1), were examined in 3T3-L1 adipocytes. PGD₂ induced a reduction in adiponectin and leptin mRNA, and the secretion of these adipokines was also inhibited, the effect being greater with leptin (up to 10-fold) than with adiponectin (twofold). In contrast, PGD₂ induced a marked stimulation of IL-6 and MCP-1 expression; with IL-6, this was rapid, the mRNA level increasing by >50-fold by 1 h. The rise in mRNA was accompanied by an increase in IL-6 and MCP-1 release (up to 100- and 6.5-fold, respectively). The effects of PGD₂ were generally mirrored by PGJ₂ and Δ¹²-PGJ₂; Δ¹²-PGJ₂ was a particularly strong stimulator of IL-6 production. These results indicate that PGD₂ and the J₂-series prostaglandins PGJ₂ and Δ¹²-PGJ₂ can have major effects on the synthesis and release of key adipokines. Such effects could be important in the inflammatory response in adipose tissue.     

Modulation of cytotoxic T lymphocyte, natural killer cell, antibody-dependent cellular cytotoxicity, and antibody-dependent complement-mediated cytotoxicity by Vernonia cinerea L. and vernolide-A in BALB/c mice via enhanced production of cytokines IL-2 and IFN-γ

Effect of Vernonia cinerea L. and vernolide-A on cell-mediated immune (CMI) response was studied in normal as well as tumor-bearing BALB/c mice. Administration of V. cinerea and vernolide-A significantly enhanced natural killer (NK) cell activity in both normal as well as tumor-bearing animals, and the activity was observed earlier than in tumor-bearing control animals. Antibody-dependent cellular cytotoxicity (ADCC) and antibody-dependent complement-mediated cytotoxicity (ACC) were also enhanced significantly in both normal as well as tumor-bearing animals after V. cinerea and vernolide-A administration compared with untreated control tumor-bearing animals. Extract and vernolide-A showed a significant increase in cytotoxic T lymphocyte (CTL) production in both the in vivo and in vitro models. The level of cytokines such as interleukin (IL)-2 and interferon (IFN)-γ were also enhanced by the treatment of V. cinerea and vernolide-A in both normal as well as tumor-bearing animals. This study demonstrated that V. cinerea extract and vernolide-A stimulate the CTL, NK cell, ADCC, and ADCC through enhanced secretion of IL-2 and IFN-γ.