Astrocyte-Derived CCL2 is Associated with M1 Activation and Recruitment of Cultured Microglial Cells

Background/Aims: Microglia are an essential player in central nervous system inflammation. Recent studies have demonstrated that the astrocytic chemokine, CCL2, is associated with microglial activation in vivo. However, CCL2-induced microglial activation has not yet been studied in vitro. The purpose of the current study was to understand the role of astrocyte-derived CCL2 in microglial activation and to elucidate the underlying mechanism(s). Methods: Primary astrocytes were pre-treated with CCL2 siRNA and stimulated with TNF-α. The culture medium (CM) was collected and added to cultures of microglia, which were incubated with and without CCR2 inhibitor. Microglial cells were analyzed by quantitative RT-PCR to determine whether they polarized to the M1 or M2 state. Microglial migratory ability was assessed by transwell migration assay. Results: TNF-α stimulated the release of CCL2 from astrocytes, even if the culture media containing TNF-α was replaced with fresh media after 3 h. CM from TNF-α-stimulated astrocytes successfully induced microglial activation, which was ascertained by increased activation of M1 and enhanced migration ability. In contrast, CM from astrocytes pretreated with CCL2 siRNA showed no effect on microglial activation, compared to controls. Additionally, microglia pre-treated with RS102895, a CCR2 inhibitor, were resistant to activation by CM from TNF-α-stimulated astrocytes. Conclusion: This study demonstrates that the CCL2/CCR2 pathway of astrocyte-induced microglial activation is associated with M1 polarization and enhanced migration ability, indicating that this pathway could be a useful target to ameliorate inflammation in the central nervous system

Effects of water temperature on growth of invasive Myriophyllum aquaticum species

This study sought to investigate the invasive mechanism of Myriophyllum aquaticum by subjecting it to simulation experiments in varying water temperatures ranging from 0 °C to 30 °C. The results showed that water temperature considerably affected both the growth and reproduction of M. aquaticum. The optimal temperature range for the growth of M. aquaticum was 25‒30 °C. Although the growth of M. aquaticum was inhibited at temperatures between 0‒5 °C, this did not result in mortality. The stem nodes, branches, and diameter reached maximum values over a temperature range of 20‒25 °C. High-temperature stress at 30 °C led to a gradual decrease or disappearance of branches. Compared to the 0 °C, 5 °C, and 30 °C treatment groups, a temperature of 20 °C led to biomass accumulation and significantly higher values. M. aquaticum’s physiological activities were affected by temperature. Except for 10 °C and 15 °C, the catalase activity varied among different water temperatures. M. aquaticum catalase activity was maximal at 5 °C and minimal at 25 °C. Conversely, the synthesis of photosynthetic pigments was highest at 10 °C and 15 °C. The plant’s optimal temperature for growth was between 20 °C and 25 °C. When the temperature was <10 °C, M. aquaticum adapted to the water temperature’s potential damage. This plant has a notable ability to tolerate various temperatures

Pre-Activation of Toll-Like Receptor 2 Enhances CD8+ T-Cell Responses and Accelerates Hepatitis B Virus Clearance in the Mouse Models

Toll-like receptors (TLRs) play a crucial role in activation of innate immunity, which is essential for inducing effective adaptive immune responses. Our previous studies have shown that toll-like receptor 2 (TLR2) is required to induce effective virus-specific T-cell responses against hepatitis B virus (HBV) in vivo. However, the contribution of TLR2 activation to adaptive immunity and HBV clearance remains to be clarified. In this study, we explored the hydrodynamic injection (HI) mouse models for HBV infection and examined how the TLR2 agonist Pam3CSK (P3C) influences HBV control and modulates HBV-specific T-cell response if applied in vivo. We found that TLR2 activation by P3C injection leads to the rapid but transient production of serum proinflammatory factors interleukin-6 and tumor necrosis factor-α and activation of CD8+ T cells in vivo. Then, the anti-HBV effect and HBV-specific T-cell immunity were investigated by TLR2 activation in the mouse models for persistent or acute HBV infections using HBV plasmids pAAV-HBV1.2 and pSM2, respectively. Both P3C application at early stage and pre-activation promoted HBV clearance, while only TLR2 pre-activation enhanced HBV-specific T-cell response in the liver. In the mouse model for acute HBV infection, P3C application had no significant effect on HBV clearance though P3C significantly enhanced the HBV-specific T-cell response. Collectively, TLR2 pre-activation enhances HBV-specific T-cell responses and accelerates HBV clearance in HI mouse models. Thus, the modulation of host immune status by TLR2 agonists may be explored for immunotherapeutic strategies to control HBV infection

Additional file 1: of Chinese woodchucks with different susceptibility to WHV infection differ in their genetic background exemplified by cytochrome B and MHC-DRB molecules

Table S1. Basic information. Table S2. Primers used for PCR and RT-PCR. Table S3. The parameters of PCR reaction mix and the amplification cycles. Table S4. Distribution of cytochrome B alleles in Chinese woodchucks. Table S5. Distribution of MHC-DRB alleles in Chinese woodchucks. (ZIP 41 kb