IFN-γ和IFN-α在IL-12诱导的TH1发展中的作用

[英]／CythiaAW／JImmunol．-1996，156．－1442～1447T辅助细胞表型的发育可能是对特异性病原因子抵抗的应急反应。在鼠实验性leishmaniasis中TH1应答的发展赋以对病原因子的抵抗力，而TH2应答则给以致死性病原因子的传播。IL－12，IL－4分别对天然（naive）T细胞向TH1和TH2亚     

HLA-G inhibits xenogenetic cytotoxicity mediated by human NK cells and T lymphocytes against PECs

The worldwide shortage in the supply of human do-nor organs is becoming more and more pronounced. Xenotransplantation may probably give the hope to over-come the problem ultimately. Because it has a great pros-pect of clinical application, xenotransplantation has drawn great attention[1]. The pig appears to be an ideal source for human transplantation. But the xenograft has to face the challenge of three severe rejections (the hyperacute rejec-tion, the delayed xenograft rejection and acute ce…     

HLA－G inhibits xenogenetic cytotoxicity mediated by human NK cells and T lymphocytes against PECs

In order to investigate whether the non-classical HLA-G class I molecule protects the prcine endothelial cells(PECs)from the lysis mediated by human immune cells in pig to human discordant xenotransplantation,we have cloned HLA-G cDNA from a human placents by RT-PCR.Mammalian expression vector,pEFG-neo,was constructed by insertion of HLA-G cDNA in pEF-neo.We obtained efficiently expressed PECs by stable transfection.Cytotoxicity assay showed that overexpression of HLA-G on PECs was sufficient to inhibit human NK-92 cell lysis.The level of lysis was equal to or less than that of the lysis of human umbilical vein endothelial cells mediated by human NK-92 cells.It also indicated that HLA-G inhibited the lysis of PECs mediated by xeno-antigen specific T lymphocytes.The reduction of lysis ranged between 59.1% and 88.9A%.These findings suggest that the transgenic approach to overexpress HLA-G is believed to be a new immunotherapy in overconing the immune rejections in xenotransplantion,including delayed xenograft rejection and cell-mediated rejection.     

Expression of human clotting factor Ⅸ mediated by recom- binant lentiviral vector in cultured cells and hemophilia B mice

Hemophilia B, a serious bleeding disorder, is an inherited X chromosome-linked disease for the deficiency or inactivity of human clotting factor Ⅸ (hFⅨ). Though factor substitution therapy has greatly improved the lives of hemophiliac patients, there are still limitations to the current treatment, which have triggered interest in alternative treatments by gene therapy[1]. Based on preclinical studies in rabbits[2], our lab had first initiated an ex vivo gene therapy clinical trial whereby a…     

Expression of human clotting factor Ⅸ mediated by recom- binant lentiviral vector in cultured cells and hemophilia B mice

To explore the expression of human clotting factor Ⅸ (hFⅨ) cDNA in vitro and the feasibility of gene therapy for hemophilia B mice mediated by recombinant lentiviral vector, a recombinant hFⅨ lentiviral vector driven by ubiquitin-C promoter, FUXW, and by ABP liver specific promoter, FAXW, was constructed respectively. Recombinant lentivirus was harvested from 293T cells by calcium phosphate-mediated transient cotransfection of three plasmids (transgene vector, CMV腞8.2, VSV-G). hFⅨ expression was detected in supernatant of 293T, BHK and L-02 cells infected with FUXW virus, whereas higher expression of hFⅨ levels (630 ng/106 cells/48 h) was detected only in L-02 cells infected with FAXW virus. Serum hFⅨ antigen was detected in all hemophilia B mice treated with FAXW virus by tail vein injection, an efficiency level of hFⅨ was observed (45 ng/mL, approximately 1% of normal human levels), the expression lasted for more than 60 d. The results indicated that HIV-based lentiviral vectors offer a promising approach to the gene therapy of hemophilia B.     

Induction of human β-defensin-2 in airway epithelial cells by Streptococcus pneumoniae:involvement of IP3-dependent intracellular calcium release and NF-jB

Human β-definsin-2(hBD-2)is mainly induced by bacterial factors and pro-inflammation mediators in epithelial cells.As the major cause of community-acquired pneumonia,whether Streptococcus pneumoniae(S.pneumoniae)stimulation induces hBD-2 expression in airway epithelial cells is elusive.In this study,we found that S.pneumoniae stimulation induced hBD-2 expression in a time-and concentration-dependent manner in primary human airway epithelial cells.To further reveal the mechanism of S.pneumoniae inducing hBD-2,we found that S.pneumoniae stimulation activated NF-jB signaling pathway.Specific NF-jB inhibitor,PDTC,could reverse the induction of hBD-2 by S.pneumoniae.We also found that cellular inner Ca2+ signaling is involved in the S.pneumoniae-induced hBD-2.Taken together,our findings indicated that S.pneumoniae can stimulate the expression of hBD-2 in airway epithelial cells and NF-jB and inositol triphosphate-dependent intracellular calcium release is involved in this induction.     

Differentiation of bone mar-row derived Thy-1~+β_2M~- cells into hepatocytes induced by coculture with transgenic CFSCs

~~Differentiation of bone mar-row derived Thy-1~+β_2M~- cells into hepatocytes induced by coculture with transgenic CFSCs~~     

Differentiation of bone mar-row derived Thy-1~+β_2M~- cells into hepatocytes induced by coculture with transgenic CFSCs

Studies of transplantation in vivo indicted thatbone marrow derived stem cells had a potential to differenti-ate into mature hepatocytes. However, there are lots ofdoubts and uncertainties in the influencing factors and con-trol agents of effectively inducing stem cell differentiation invitro, the efficiency of stem cells‘ differentiation into hepato-cytes and differentiated cells‘ life-span and functional state,etc. In this study, rat bone marrow derived Thy-l^ β2M^- cells(BDTCs) were induced to differentiate into hepatocytes byco-culturing with CFSC/HGF feeder layers which expressedhHGF efficiently and stably. RT-PCR and immunofluores-cent texts proved induced BDTCs expressed infant and adulthepatocyte specific genes. Further more, these cells displayedfunctions of indocyanine green (ICG) uptake, ammoniummetabolism and albumin production. It was shown thatgrowth factors together with hepatic nonparenchyma cellsprovided a feasible microenvironment for differentiation ofbone marrow stem cells into hepatocytes. The studies notonly provided a significant biological model for going deepinto the mechanism of stem cell plasticity, but also offered atheoretical and technical foundation of gene and stem cellengineering-based regenerative medicine for end-stage liverdiseases.     

Molecular cloning and expression of interleukin Ibeta (IL-1β) from red seabream (Pagrus major)

The interleukin 1β (IL-1β) cDNA was cloned from the red seabream (Pagrus major) by homology cloning strategy. A cDNA fragment was amplified by PCR using two degenerated primers, which were designed according to the conserved regions of other known IL-1β sequences, and elongated by 3' ends and 5' ends RACE PCR to get the full length coding sequence of red seabream IL-1β (RS IL-1β). The sequence contained 1252 nucleotides that included a 5' untranslated region (UTR) of 84bp, a 3' UTR of 410 bp and an open reading frame (ORF) of 759 nucleotides which could be translated into a putative peptide of 253 amino acids with molecular weight of 28.6 kD and putative isoelectric point pI of 5.29. The deduced peptide contained two potential N-glycosylation sites and an identifiable IL1 family signature, but lacked the signal peptide and the clear ICE cut site, which were common in other nonmammalian IL-1β genes. The RS IL-1β had the highest homology with piscine IL-1β according to phylogenetic tree analysis.     

Induction of human β-defensin-2 in airway epithelial cells by Streptococcus pneumoniae： involvement of IP3-dependent intracellular calcium release and NF-κB

Human β-definsin-2 （hBD-2） is mainly induced by bacterial factors and pro-inflammation mediators in epithelial cells. As the major cause of community-acquired pneumonia, whether Streptococcus pneumoniae （S. pneu-moniae） stimulation induces hBD-2 expression in airway epithelial cells is elusive. In this study, we found that S. pneumoniae stimulation induced hBD-2 expression in a time-and concentration-dependent manner in primary human airway epithelial cells. To further reveal the mechanism of S. pneumoniae inducing hBD-2, we found that S. pneumoniae stimulation activated NF-κB signaling pathway. Specific NF-κB inhibitor, PDTC, could reverse the induction of hBD-2 by S. pneumoniae. We also found that cellular inner Ca^2＋ signaling is involved in the S. pneumoniae-induced hBD-2. Taken together, our find-ings indicated that S. pneumoniae can stimulate the expression of hBD-2 in airway epithelial cells and NF-κB and inositol triphosphate-dependent intracellular calcium release is involved in this induction.     

Effect of P15INK4b expression on the cell cycle and G1 phase related regulatory genes in human melanoma cells

Using the transfeetion teehnique. P15INK4b was introduced into P15INk4b gene deleted human melanoma A375 cells,and a cell model MLED6 overexpressing P15INK4b WAS CONSTRUCTED.Comparing with the control cells MLC2,MLEK6cells in G1phase increased by 11%,but those in Sphase decreased by 15%by FCM.By the method of thymidine(TdR)and N2O arresting,the proportions of synchronized Mphase cells of MLEK6 ana MLC23 were measured and found to be 89.1% and 76.8%respectively ,and the cells in G1phase were 74.3% for MLID6 AND 76. 4% forMLC2.The result of3 H-TdR incorporation indicated that the transition of G1/Sof MLEK6 cell was delayed 2h as compared with that of MLC2 cells,and incorporation rate also decreased.The observation on exprissions of some G1/ S-resates relatory rigusating genes showed that in MLIK6 cells the protein leves of P27KIPI increased with the decreasing expressions of cyclinD1,cyclinE and c-myc,especially cyclinD1 in late G1phade.The expression of cyclinE obviously decreased at G1/S transition ,and c-myc wad inhibited throughout all the process of G1 S phase.All the risults suggest that P15INK4b can delayG1/S transition of MLEK6 cells by inhibiting the cell cycle engine ,and by increasing the expression of Cdk ingibitor P27KIPI in different stages of G1 phase.     

幼兔和成年兔感染兔出血热病毒后血清中IL-8、IL-10、TNF-α、TGF-β和MCP-1检测分析

兔出血症病毒(RHDV)是兔出血症的病原体,可导致成年兔大量死亡.研究表明,细胞因子在RHD致病中发挥了重要作用.为探索细胞因子在RHDV对幼兔和成年兔致病差异性中的作用机制,通过对幼兔(2周龄)和成年兔(3月龄)肌肉注射检测为RHDV核酸阳性的病兔肝脏悬液,酶联免疫吸附试验(ELISA)检测感染RHDV后不同时间(0、12、24、36、48、60 h)血清中IL-8、IL-10、TNF-α、TGF-β和MCP-1的水平.ELISA结果显示,成年兔感染RHDV后血清中IL-8、IL-10、TNF-α、TGF-β和MCP-1水平在0～36 h显著升高(P<0.05),并在感染后36～48 h死亡.幼兔感染后血清中IL-8水平在24～60 h明显上升(P<0.05),TNF-α和TGF-β水平变化无明显规律,IL-10和MCP-1水平无明显变化,且未出现死亡现象.比较幼兔和成年兔上述细胞因子水平发现,成年兔感染后血清中IL-8、IL-10、TGF-β和MCP-1水平显著高于幼兔(P<0.05),TNF-α水平在感染后24 h左右高于幼兔(P<0.001),其余时间均...     

Effect of Aβ1-40 on membrane permeability and intracellular free Ca2+ of nerve cells

The effects of soluble and fibrillar Aβ1-40 on membrane permeability and intracellular free Ca^2 of nerve cells were investigated by the laser confocal microscopy. Results indicate that: i) Effects of soluble and fibrillar Aβ1-40 on cell membrane permeability are both concentration-dependent. Soluble Aβ1-40 increases membrane permeability only at concentration of 3μmol/L, while the toxic effect of fibrillar Aβ1-40 is much stronger, its evident effect begins from 1μmol/L. When its concentration rose to 3μmol/L, not only the membrane permeability increased, but also the nuclear membrane broke seriously, ii) Both soluble and fibrillar Aβ1-40 at high concentrations increased the intracellular free Ca^2 , and the increased amplitudes are concentration-dependent. However, the fibrillar one induces the increase of intracellular Ca^2 much quicker and synchronously.These results indicate that some correlation exists between the neurotoxicity of high concentration soluble and fibrillar Aβ1-40 and the change of physico-chemical properties and intracellular Ca ion imbalance.     

1-C-(2''''-Oxoalkyl)-Glycosides as Latent α,β-Unsaturated Conjugates. Synthesis of Aza-C-glycosides and Thio-C-Glycosides by an Intramolecular Hetero-michael Addition

1 Introduction Aza-sugars, a class of polyhydroxylated pyrrolidine and piperidine, are powerful glyco-processing enzyme inhibitors and potential therapeutics for the treatment of diabetic, cancer, viral infection and other diseases~([1]). For example, N-hydroxyethyl-1-deoxynojirimycin (Miglitol) and N-butyl-1-deoxynojirimycin (Zavestca) have been approved for the treatment of type 2 diabetes and type 1 Gaucher disease, respectively. Numerous synthetic methods towards azasugars have been deve…