Double-negative T cells remarkably promote neuroinflammation after ischemic stroke.

CD3+CD4-CD8- T cells (double-negative T cells; DNTs) have diverse functions in peripheral immune-related diseases by regulating immunological and inflammatory homeostasis. However, the functions of DNTs in the central nervous system remain unknown. Here, we found that the levels of DNTs were dramatically increased in both the brain and peripheral blood of stroke patients and in a mouse model in a time-dependent manner. The infiltrating DNTs enhanced cerebral immune and inflammatory responses and exacerbated ischemic brain injury by modulating the FasL/PTPN2/TNF-α signaling pathway. Blockade of this pathway limited DNT-mediated neuroinflammation and improved the outcomes of stroke. Our results identified a critical function of DNTs in the ischemic brain, suggesting that this unique population serves as an attractive target for the treatment of ischemic stroke.

Effect of low frequency magnetic fields on melanoma: tumor inhibition and immune modulation.

BACKGROUND: We previously found that the low frequency magnetic fields (LF-MF) inhibited gastric and lung cancer cell growth. We suppose that exposure to LF-MF may modulate immune function so as to inhibit tumor. We here investigated whether LF-MF can inhibit the proliferation and metastasis of melanoma and influence immune function. METHODS: The effect of MF on the proliferation, cell cycle and ultrastracture of B16-F10 in vitro was detected by cell counting Kit-8 assay, flow cytometry, and transmission electron microscopy. Lung metastasis mice were prepared by injection of 2 × 105 B16-F10 melanoma cells into the tail vein in C57BL/6 mice. The mice were then exposed to an LF-MF (0.4 T, 7.5 Hz) for 43 days. Survival rate, tumor markers and the innate and adaptive immune parameters were measured. RESULTS: The growth of B16-F10 cells was inhibited after exposure to the LF-MF. The inhibition was related to induction of cell cycle arrest and decomposition of chromatins. Moreover, the LF-MF prolonged the mouse survival rate and inhibited the proliferation of B16-F10 in melanoma metastasis mice model. Furthermore, the LF-MF modulated the immune response via regulation of immune cells and cytokine production. In addition, the number of Treg cells was decreased in mice with the LF-MF exposure, while the numbers of T cells as well as dendritic cells were significantly increased. CONCLUSION: LF-MF inhibited the growth and metastasis of melanoma cancer cells and improved immune function of tumor-bearing mice. This suggests that the inhibition may be attributed to modulation of LF-MF on immune function and LF-MF may be a potential therapy for treatment of melanoma.

Ampelopsin attenuates lipopolysaccharide-induced inflammatory response through the inhibition of the NF-κB and JAK2/STAT3 signaling pathways in microglia.

Increasing evidence suggests that microglia are a major cellular contributor to neuroinflammation. The present study investigated whether Ampelopsin (Amp), a type of flavanonol derivative from Ampelopsis grossedentata, may exert an anti-inflammatory effect on lipopolysaccharide (LPS)-induced BV2 and primary microglia cells. We found that pre-treatment of microglia cells with Amp before LPS with a non-cytotoxic concentration range decreased the production of nitric oxide (NO) and prostaglandin E2 (PGE2). Amp also suppressed the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) at the mRNA and protein levels. In addition, LPS-induced production of pro-inflammatory cytokines such as interleukin (IL)-1ß, IL-6, and tumor necrosis factor-α (TNF-α) was obviously reduced by Amp. Our mechanistic study indicated that Amp suppressed LPS-induced activation of the IκB/NF-κB inflammation pathway without affecting changes in the phosphorylation levels of mitogen-activated protein kinases (MAPKs) in BV2 cells. Further studies revealed that Amp markedly reduced the phosphorylation levels of JAK2-STAT3 and STAT3 nuclear translocation. Overall, our data suggest that Amp can suppress the LPS-induced inflammatory response of microglial cells, indicating that Amp has potential for the treatment of inflammation-mediated neurodegenerative diseases.

Association of increased Treg and Th17 with pathogenesis of moyamoya disease.

Immuno-inflammation has been shown to play a pivotal role in the pathogenesis of moyamoya disease (MMD). However, how did circulating Treg/Th17 cells involve in MMD patients remains unclear. 26 MMD, 21 atherothrombotic stroke, and 32 healthy controls were enrolled in this study. MMD patients have a significantly higher percentage of circulating Treg and Th17 cells as well as their dominantly secreting cytokines than other groups (P < 0.0001), whereas no difference was found in the ratio of Treg/Th17 between patients in MMD and atherothrombotic stroke group or control subjects (P = 0.244). However, the increased Treg in MMD patients which were enriched with FrIII Treg cells had deficient suppressive functions (P = 0.0017) compared to healthy volunteers. There was a positive correlation between Treg or TGF-ß and MMD Suzuki's stage. And the level of circulating Treg was as an independent factor associated with MMD stage. Besides, TGF-ß was also correlated with the increased expression of VEGF in MMD patients. Our findings indicated an important involvement of circulating Treg in the pathogenic development of MMD and TGF-ß in Treg induced VEGF.

Malibatol A enhances alternative activation of microglia by inhibiting phosphorylation of Mammalian Ste20-like kinase1 in OGD-BV-2 cells.

OBJECTIVE: To investigate the polarization effect of Malibatol A on oxygen-glucose deprivation (OGD)-BV-2 cells, and the possible molecular mechanism involved in c-Abl-MST signaling pathway. METHOD: The OGD BV-2 cell model was established. BV-2 cells were exposed to OGD for 8 h followed by reperfusion for 15 h with Malibatol A at different concentration of 0.5, 1, 2, 4, 8, 16 µM or without it. And then cells, mRNA and protein were harvested respectively. The cell viability and apoptosis were measured by MTT assay and flow cytometry. The mRNA of classical activated microglia (M1) markers (MCP-1, IL-1 and TNF-α) and alternatively activated microglia (M2) markers (Ym-1, CD206, IL-10, TGF-ß) in BV-2 cells were measured by RT-PCR. Meanwhile, the proteins of Ym-1 and CD206 was assayed by flow cytometry. Furthermore, the expression of c-Abl and MST was measured by Western blot. RESULT: Malibatol A significantly decreased apoptosis and increased viability of OGD BV-2 cells in a dose-dependent manner. In the presence of Malibatol A, the mRNA levels of Ym-1, CD206, IL-10 and TGF-ß mRNA was significantly increased in OGD-BV-2 cells, while the mRNA levels of MCP-1, IL-1 and TNF-α was obviously down-regulated. Meanwhile, the proteins of Ym-1 and CD206 was raised in OGD BV-2 cells with Malibatol A. Besides, Malibatol A also inhibited OGD-induced p-MST1(Y433) in BV-2 cells. CONCLUSION: Malibatol A could attenuate OGD-induced BV-2 cell injury and promote M2 microglia polarization. The mechanism may be related to inhibition of MST1 phosphorylation at Y433.

Increased Expressions of Plasma Galectin-3 in Patients with Amyotrophic Lateral Sclerosis.

BACKGROUND: High expressions of galectin-3 were identified recently in the end stage of amyotrophic lateral sclerosis (ALS) patients, which suggested that immune reactivity and inflammatory mechanisms might play an important role in the pathogenesis of ALS. The purpose of this study was to investigate plasma galectin-3 levels in different groups and stages of ALS patients and the association with related clinical characteristics. METHODS: A total of 51 patients with ALS and 60 normal controls (NCs) were recruited in this study. Plasma galectin-3 levels were determined using the enzyme-linked immunosorbent assay. Patients with ALS were divided into several groups according to their clinical characteristics: gender, type of disease onset, duration of disease, and clinical conditions of disease. Statistical analyses of the differences of galectin-3 levels between groups and the association with the clinical characteristics of disease were performed. RESULTS: As compared with the NCs (201.64 [22.35-401.63] ng/ml), plasma galectin-3 levels were significantly elevated in the patients with duration >12 months (341.17 [69.12-859.22] ng/ml, P< 0.05), and the patients with limb onset of disease (254.14 [69.12-859.22] ng/ml, P< 0.05); however, no difference was found in the patients with duration ≤12 months (250.62 [109.77-334.92] ng/ml, P > 0.05), and the patients with bulbar onset of disease (251.79 [109.20-404.76] ng/ml, P > 0.05). In addition, galectin-3 levels were significantly increased in the female patients (263.27 [123.32-859.22] ng/ml, P< 0.05) while no difference was found in the male patients (220.39 [69.12-748.73] ng/ml, P > 0.05). The further statistical analyses showed that plasma galectin-3 levels were positively correlated with the duration of disease (r = 0.293, P = 0.037). CONCLUSIONS: Plasma galectin-3 levels were significantly increased in ALS patients with limb onset of disease, especially in ALS female patients, and positively correlated with the duration of disease, which suggested that plasma galectin-3 might be an interesting and useful factor associated with ALS.

Human umbilical cord mesenchymal stem cells protect against ischemic brain injury in mouse by regulating peripheral immunoinflammation.

Current treatments for ischemic stroke are limited, stem cell transplantation offers great potential as a therapeutic strategy. The present study was undertaken to determine whether human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) could improve brain injury after middle cerebral artery occlusion (MCAO) through modulating peripheral immunoinflammation. The study showed that neurological deficit was ameliorated and brain edema, infarct volume was significantly decreased from 72 h to 1 week post-MCAO with hUC-MSCs treatment via tail vein injection within 30 mins after stroke; hUC-MSCs attenuated the levels of inflammatory factors including IL-1, TNF-α, IL-23, IL-17 and IL-10 in peripheral blood serum and ischemia hemisphere after stroke; hUC-MSCs significantly decreased the level of Th17 cells at 24h and increased the level of Tregs at 72 h post-MCAO in peripheral immune system; the level of TGF-ß in blood serum was enhanced by hUC-MSCs. In conclusion, our findings suggested that hUC-MSCs had neuroprotection in MCAO mice by TGF-ß modulating peripheral immune and hUC-MSCs may be as a potential therapy for ischemic stroke.

Low frequency magnetic fields enhance antitumor immune response against mouse H22 hepatocellular carcinoma.

OBJECTIVE: Many studies have shown that magnetic fields (MF) inhibit tumor growth and influence the function of immune system. However, the effect of MF on mechanism of immunological function in tumor-bearing mice is still unclear. METHODS: In this study, tumor-bearing mice were prepared by subcutaneously inoculating Balb/c mice with hepatocarcinoma cell line H22. The mice were then exposed to a low frequency MF (0.4 T, 7.5 Hz) for 30 days. Survival rate, tumor growth and the innate and adaptive immune parameters were measured. RESULTS: MF treatment could prolong survival time (n = 28, p<0.05) and inhibit tumor growth (n = 9, p<0.01) in tumor-bearing mice. Moreover, this MF suppressed tumor-induced production of cytokines including interleukin-6 (IL-6), granulocyte colony- stimulating factor (G-CSF) and keratinocyte-derived chemokine (KC) (n = 9-10, p<0.05 or 0.01). Furthermore, MF exposure was associated with activation of macrophages and dendritic cells, enhanced profiles of CD4(+) T and CD8(+) T lymphocytes, the balance of Th17/Treg and reduced inhibitory function of Treg cells (n = 9-10, p<0.05 or 0.01) in the mice model. CONCLUSION: The inhibitory effect of MF on tumor growth was related to the improvement of immune function in the tumor-bearing mice.