PMC:4277126 / 90480-99549 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/4277126","sourcedb":"PMC","sourceid":"4277126","source_url":"http://www.ncbi.nlm.nih.gov/pmc/4277126","text":"T cells, B cells, and splenocytes\nChen et al120 compared the immunomodulating effects of different LBP fractions in mice. Crude LBPs isolated from L. barbarum were separated to obtain five homogeneous fractions, namely LBPF1, LBPF2, LBPF3, LBPF4, and LBPF5. The study showed that LBP, LBPF4, and LBPF5 significantly stimulated mouse splenocyte proliferation. The proliferation proved to be of T cells, but not B cells. Cell cycle analysis indicated that LBP, LBPF4, and LBPF5 markedly reduced sub-G1 cells.120 LBP, LBPF4, and LBPF5 activated the transcription factors nuclear factor of activated T-cells (NFAT) and activator protein-1 (AP-1), prompted CD25 (ie, IL-2 receptor-α) expression, and induced IL-2 and interferon (IFN)-γ expressions. NFAT proteins (NFATs 1–5) have crucial roles in the development and function of the immune system. In T cells, NFAT proteins not only regulate activation but are also involved in the control of thymocyte development, T-cell differentiation, and self-tolerance.121 AP-1 regulates gene expression in response to a variety of stimuli, including cytokines, growth factors, stress, and bacterial and viral infections. IL-2 is important for the growth and activation of T cells, and IFN-γ is an important activator of macrophages and inducer of class II major histocompatibility complex (MHC-II) molecule expression. IL-2 is mainly produced by T cells122 and IFN-γ is produced predominantly by NK and NK T cells as part of the innate immune response, and by CD4+ T helper cells (Th1) and CD8+ CTLs once antigen-specific immune response is triggered.123 Administration of LBPs to mice (intraperitoneal [ip] or oral administration [po]) significantly induced T-cell proliferation.120 These results suggest that activation of T lymphocytes by LBPs may contribute to one of their immuno-enhancement functions.\nThe in vitro and in vivo immunomodulating effects of LBPF4-OL on mouse splenocytes, T cells, B cells, and macrophages were investigated by Zhang et al.124 LBPF4-OL was the glycan part of L. barbarum polysaccharide–protein complex fraction 4 (LBPF4). Splenocytes were stimulated with LBPF4-OL and cytokine concentrations in the supernatants were determined. In the in vivo study, mice were intraperitoneally injected with 100 µg/mL LBPF4-OL daily for 6 days. The results showed that LBPF4-OL markedly induced the splenocyte proliferation, but could not induce proliferation of purified T- and B-lymphocytes.124 B-cell proliferation occurred in the presence of activated macrophages or lipopolysaccharide (LPS). LBPs obviously induced IL-6, IL-8, IL-10, and TNF-α production in splenocytes in a concentration-dependent manner.124 IL-6 is secreted by T cells and macrophages to stimulate immune response during infection and after trauma (especially burns or other tissue damage) leading to inflammation. IL-8 (also called CXCL8 and neutrophil chemotactic factor) is a chemokine produced by macrophages and other cell types such as epithelial cells, airway smooth muscle cells, and endothelial cells. IL-8 induces chemotaxis in target cells (primarily neutrophils but also other granulocytes) toward the site of infection and induces phagocytosis. IL-10 inhibits the production of IFN-γ, IL-2, IL-3, TNF-α, and granulocyte-macrophage colony-stimulating factor (GM-CSF) by activated macrophages and helper T cells. TNF-α is mainly produced by activated macrophages (M1), although it can be produced by many other cell types such as CD4+ T lymphocytes, monocytes, NK cells, neutrophils, mast cells, eosinophils, and neurons.125 The primary role of TNF-α is in the regulation of immune cells. TNF-α induces fever, apoptotic cell death, cachexia, and inflammation; inhibits tumorigenesis and viral replication; and responds to sepsis via IL-1 and IL-6 producing cells.125 Flow cytometer analysis showed that LBPF4-OL prompted CD86 (B7-2) and MHC-II molecule expression on macrophages and greatly promoted release of TNF-α and IL-1β from macrophages.124 IL-1β is produced by activated macrophages as a proprotein, which is proteolytically processed to its active form by caspase 1. This cytokine is an important mediator of the inflammatory response and is involved in a variety of cellular activities, including cell proliferation, differentiation, and apoptosis.\nVidal et al126 revealed that dietary wolfberry supplementation enhanced both in vivo (delayed-type hypersensitivity) and ex vivo (T-cell proliferation) T-cell response to specific antigens, but it did not affect mitogen-induced T-cell or B-cell proliferation in young and aged mice. Over 44 days, young-adult (2 months) and aged (21 months) C57BL/6J mice were fed ad libitum with a controlled diet and received drinking water supplemented or not with 0.5% (wt/vol) Lacto-Wolfberry. All mice were immunized on day 15 and challenged on day 22 with a T-cell-dependent antigen, keyhole limpet hemocyanin. The study showed that Lacto-Wolfberry supplementation significantly increased in vivo systemic immune markers.126 Both antigen-specific humoral response and cell-mediated immune responses in young-adult and aged mice were enhanced. However, no significant effect of Lacto-Wolfberry supplementation was observed on ex vivo splenocyte proliferative response to mitogens and on splenocyte T-cell subsets.126 These data suggest that dietary intake of Lacto-Wolfberry may favorably modulate the poor responsiveness to antigenic challenge observed with aging.\nZhang et al127 further compared the effect of LBPF4 and LBPF4-OL on the proliferation of splenocytes and mitogen-induced B and T lymphocytes in female Balb/C mice. LBPF4 and LBPF4-OL were isolated in the fruit bodies of L. barbarum through a series of diethylaminoethyl anion exchange cellulose and gel-permeation chromatography. The molecular weight of LBPF4 was 214.8 kDa, and consisting of 17 amino acids and four kinds of monosaccharides. The molecular weight of LBPF4-OL was 181 kDa, consisting of three types of monosaccharides.127 The effects on cytokine secretion, the phagocytic potential of macrophages, and the expression level of intracellular signaling molecules including NF-κB and B-cell-specific activator protein (BSAP, also named Pax5) were also determined. BSAP/Pax5 is essential for commitment of lymphoid progenitors to the B lymphocyte lineage.128 Spleen cells (5×105) were stimulated with 10 µg/mL, 50 µg/mL, and 100 µg/mL LBPF4-OL. Concanavalin A (Con A, 0.5 µg/mL) and LPS (5 µg/mL) were included as positive controls for the proliferation of T and B cells, respectively. The results showed that 50 µg/mL LBPF4 significantly enhanced spleen cell proliferation ∼3.2 fold, while LBPF4-OL enhanced proliferation 7.2 fold. Administration of 10 µg/mL, 50 µg/mL, or 100 µg/mL LBPF4 but not LBPF4-OL, significantly enhanced the Con A-induced T lymphocyte proliferation.127 However, LPS-induced B-cell proliferation was enhanced by 10 µg/mL, 50 µg/mL, or 100 µg/mL of both LBPF4 and LBPF4-OL. Administration of 50 µg/mL LBPF4-OL was more effective on inducing the proliferation of splenocytes and LPS-stimulated B cells than 100 µg/mL LBPF4. LBPF4 appeared to induce lymphocyte proliferation predominantly depending on both B and T cells, and LBPF4-OL induced lymphocytes proliferation only depending on B cells. The stimulation of murine peritoneal macrophages with LBPF4 and LBPF4-OL resulted in a comparable dose-dependent increase of the production of TNF-α and IL-1β.127 In addition, both LBPF4 and LBPF4-OL at concentrations of 10 µg/mL, 50 µg/mL, and 100 µg/mL increased the secretion of NO to comparable levels. Administration of 10 µg/mL LBPF4 and LBPF4-OL showed no significant effects on the phagocytic activity of resting macrophages in mice, but the macrophage chicken erythrocyte phagocytic activity was significantly increased by low concentrations of LBPF4 and LBPF4-OL. About 50 µg/mL (but not 10 µg/mL) LBPF4 and LBPF4-OL significantly promoted BSAP and NF-κB activity.127 These data suggest that LBPF4-OL can only enhance B cell and macrophage functions, but polysaccharide–protein complex LBPF4 can enhance the function of both T and B cells and macrophages.\nRecently, Zhang et al129 found that LBPF4-OL acted as an activator of the Toll-like receptor 4 (TLR4)/p38 MAPK signaling pathway using TLR4 knockout mice. LBPF4-OL significantly induced TNF-α and IL-1β production in peritoneal macrophages isolated from wild type (C3H/HeN) but not TLR4-deficient mice (C3H/HeJ). The proliferation of LBPF4-OL-stimulated lymphocytes from C3H/HeJ mice was significantly lower than that of lymphocytes from C3H/HeN mice.129 Furthermore, through a bio-layer interferometry assay, LPS but not LBPF4-OL directly associated with the TLR4/MD2 molecular complex. Flow cytometry analysis indicated that LBPF4-OL markedly upregulated TLR4/MD2 expression in both peritoneal macrophages and Raw264.7 cells.129 LBPF4-OL also increased the phosphorylation of p38-MAPK and inhibited the phosphorylation of JNK and Erk1/2. These data suggest that LBPF4-OL can activate TLR4/p38 MAPK signaling 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