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Objective: To investigate the cytokine response pattern (IL-4/IFN-γ) in infants with RSV infections and bronchiolitis during the acute phase. Methods: Four-color flow cytometry was used to measure intracellular IL-4 and IFN-γ expressions in peripheral blood CD3+ and CD8+ lymphocytes from RSV-infected and bronchiolitis infants. Serum IL-4 and IFN-γ levels were also determined. Results: RSV-infected and bronchiolitis infants showed no statistical differences from not-RSV-infected or pneumonia infants and control in the frequency of IL-4 and IFN-γ expressions in CD3+CD8- lymphocytes, showed no obvious Th1/Th2 imbalance, while IFN-γ was expressed much more frequently in CD3+CD8+ lymphocytes. Systematically, RSV-infected and bronchiolitis infants showed much lower levels of serum IL-4 and IL-4/IFN-γ ratios and much higher serum IFN-γ levels than control. However, there were no statistical differences in the above three indices between RSV-infected and not-RSV infected infants or between bronchiolitis and pneumonia infants, except that bronchiolitis infants had a higher level of serum IFN-γ than pneumonia infants statistically. Conclusions: There is no type-2 cytokine response predominance in the acute phase of RSV infection and bronchiolitis. IL-4 production is suppressed and IFN-γ production upregulated, the latter being most prominent in bronchiolitis infants. 相似文献
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在人体内,CCR5与许多免疫疾病有关,CCR5有望成为众多药物的作用靶点。将ccr5基因与真核表达载体pBBS242构建成重组质粒pBBS242-ccr5,转染CHO细胞,并经潮霉素B筛选。流式细胞仪检测结果表明CCR5在CHO细胞得到了稳定表达。 相似文献
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Effect of perioperative autologous versus allogeneic blood transfusion on the immune system in gastric cancer patients 总被引：2，自引：0，他引：2
Background: Allogeneic blood transfusion-induced immunomodulation (TRIM) and its adverse effect on the prognosis of patients treated surgically for cancer remain complex and controversial. However, the potential risk associated with allogeneic blood transfusion has heightened interest in the use of autologous blood transfusion. In the present study, the serum concentrations of neopterin, interferon-gamma (IFN-γ), T lymphocyte subsets (CD3 , CD4 , CD8 , CD4 /CD8 ) and a possible association between these variables were investigated. The purpose was to further evaluate the effect of autologous versus allogeneic blood transfusion on immunological status in patients undergoing surgery for gastric cancer. Methods: Sixty ASA I～II (American Society of Anesthesiologists) patients undergoing elective radical resection for stomach cancer were randomly allocated to receive either allogeneic blood transfusion (n=30) or autologous blood transfusion (n=30). Serum concentrations of the neopterin, IFN-γ and T lymphocyte subsets in the recipients were measured before induction of anesthesia, after operation, and on the 5th postoperative day. Results: Both two groups, serum neopterin, IFN-γ, percentages of T-cell subsets (CD3 , CD4 ), and CD4 /CD8 ratio had significantly decreased after operation, but decreased more significantly in group H (receiving allogeneic blood transfusion) than those in group A (receiving autologous whole blood transfusion) (P<0.05). On the 5th postoperative day, serum neopterin, IFN-γ, CD3 , CD4 T-cells, and CD4 /CD8 ratio returned to the baseline values in group A. In contrast, the above remain decreasing in group H, where there were no significant relations between serum neopterin and IFN-γ. Conclusion: Perioperative surgical trauma and stress have an immunosuppressive impact on gastric cancer patients. Allogeneic blood transfusion exacerbates the impaired immune response. Autologous blood transfusion might be significantly beneficial for immune-compromised patients in the perioperative period, clearly showing its superiority over allogeneic blood transfusion. 相似文献
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Potential immunomodulation effect of the extract of Nigella sativa on ovalbumin sensitized guinea pigs 总被引：1，自引：0，他引：1
Boskabady MH Keyhanmanesh R Khameneh S Doostdar Y Khakzad MR 《Journal of Zhejiang University. Science. B》2011,12(3):201-209
Several different pharmacological effects have been described for Nigella sativa (Siah-Daneh), including an anti-inflammatory effect. In the present study, the effect of the extract of N. sativa on lung pathology and blood interleukin-4 (IL-4) and interferon-γ (IFN-γ) of sensitized guinea pigs was examined. Three groups
(n=8 for each group) of guinea pigs sensitized to ovalbumin (OA) were given drinking water alone, and drinking water containing
low and high concentrations of the plant extract, respectively. The animals of the control group (n=8) were treated with saline instead of OA and were given drinking water. The pathological changes of the lung, including
infiltration of eosinophils and lymphocytes, local epithelial necrosis, the presence of oedema, thickening of the basement
membrane, smooth muscle layer hypertrophy, mucosal secretion, and the presence of mucosal plug, and blood IL-4 and IFN-γ of
sensitized guinea pigs were evaluated. The lungs of the sensitized group showed significant pathological changes (P<0.001). Blood IL-4 and IFN-γ were increased in sensitized animals compared to the controls (P<0.01 and P<0.001, respectively). Treatment of sensitized animals with the extract led to a significant decrease in pathological changes
of the lung (P<0.01 to P<0.001), except for the oedema in the sensitized group treated with low concentration of the extract, but an increased IFN-γ.
These results confirm a preventive effect of N. sativa extract on lung inflammation of sensitized guinea pigs. 相似文献
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Transfection and expression of exogenous gene in laying hens oviduct in vitro and in vivo 总被引：2，自引：0，他引：2
Gao B Sun HC Song CY Wang ZY Chen Q Song HQ 《Journal of Zhejiang University. Science. B》2005,6(2):137-141
To examine whether or not the regulatory sequence of chicken ovalbumin gene can drive transgene expression specifically in hen oviduct, the authors constructed an oviduct-specific expression vector (pOV), containing 3.0 kilobases (kb) of the 5'-flanking sequence and 3.0 kb of the 3'-flanking sequence of the chicken 相似文献
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增殖细胞核抗原(proliferating cell nuclear antigen,PCNA),也称周期蛋白或DNA聚合酶的辅助蛋白,是真核细胞合成所必需的核蛋白,在DNA复制中起重要作用。前期实验发现日本七鳃鳗肝脏cDNA文库的表达序列标签(Expressed Sequence Tag,EST)中存在与高等脊椎动物pcna基因同源的序列。提取日本七鳃鳗(Lampetra japonica)肝脏组织RNA,通过RT-PCR方法扩增七鳃鳗pcna基因,对其进行生物信息学分析,并将Lj-pcna基因成功构建到pGFP-N2真核表达载体上,重组质粒PGFP-N2-Lj-pcna转染人Hela细胞,荧光显微镜下观察有荧光蛋白的表达。日本七鳃鳗pcna基因的真核表达载体成功构建和转染,为探讨七鳃鳗pcna基因功能研究及其它七鳃鳗相关研究提供条件。 相似文献
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基于基因工程技术 ,用PCR法扩增出编码ICA6 9的cDNA片段 ,直接克隆到pSPORT 1质粒上 ,经DNA序列测定 ,插入到GST融合蛋白表达载体 pGEX 2T ,构成重组质粒 p2T ICA6 9,得到的表达产物GST ICA6 9融合蛋白用间接ELISA法检测其免疫原性 .测序结果表明 ,所获PCR产物已正确重组到PGEX 2T表达型质粒中 .重组质粒在原核细胞中表达的融合蛋白具有免疫原性 ,并能应用于Ⅰ型糖尿病病人血清中抗ICA6 9抗体的检测 .所获得的表达产物为重组ICA6 9融合抗原 ,有助于提高Ⅰ型糖尿病的预报率和确诊率 . 相似文献
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通过PCR技术扩增出人铁蛋白基因,经酶切后与表达载体质粒pGEX-4T-2连接,重组质粒转化感受态大肠杆菌,利用菌落PCR、质粒双酶切、测序,证实成功地构建了人铁蛋白基因表达载体,利用IPTG对重组菌进行诱导表达,通过尿素洗涤纯化目的蛋白用于制备抗体． 相似文献
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从假单胞菌（Pseudomonassp．）XZG36中克隆弹性蛋白酶基因,构建原核表达载体,实现其在大肠杆菌（Escherichiacoli）中的高效表达,并对表达产物进行酶学性质分析,为微生物发酵生产弹性蛋白酶奠定基础．以假单胞菌基因组DNA为模板,PCR扩增弹性蛋白酶基因,并将其开放阅读框（0RF）克隆至融合表达载体pET30a（＋）进一步IPTG诱导表达;表达产物经His·Bind亲和层析纯化后对弹性蛋白酶进行酶学性质分析．实验成功克隆了弹性蛋白酶基因,DNA基因片段为1672bp、编码497个氨基酸残基的多肽,与预计长度相符合;实现了其在E．coli中的高效表达,表达量约占菌体总蛋白的20％;经SDS-PAGE分析,相对分子质量为48000,与预期的一致;提纯后的表达蛋白SDS-PAGE分析可见单一条带,纯度可达92％以上．表达蛋白具有良好活性． 相似文献
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将萤火虫萤光素酶基因重组到质粒pBR325中,通过“三亲交配”和同源重组,将萤光素酶基因整合到Ti质粒pGV3850:1103。通过改良的叶盘转化法,用萤光素酶基因转化单子叶植物花叶芋,获得转基因植株。对再生植株的胭脂碱测定、新霉素磷酸转移酶分析、DNA点杂交、萤光素酶基因活性测定,证明萤光素酶基因已转化到花叶芋中并进行了表达。 相似文献
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根据GenBank数据库中猪圆环病毒Ⅱ型的基因组序列设计引物,采用PCR技术从病料基因组DNA中扩增出PCVⅡ河南地方株ORF4基因,全长180 bp,编码59个氨基酸.将该基因克隆至载体pGEX-4T-3中形成pGEX-4T-3-ORF4表达载体.经PCR、酶切和测序鉴定后,转化表达菌株BL21（DE3）诱导表达.SDS-PAGE结果显示：ORF4能够在大肠杆菌中表达,产物的分子量约为32 kD,且以包涵体形式存在.Western Blot检测结果显示,纯化后的ORF4蛋白能够与鼠抗6＆#215;His标签单克隆抗体发生特异性反应,为进一步研究PCVⅡORF4蛋白的特性与功能奠定了基础. 相似文献
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抑癌基因PTEN载体构建及在LOVO细胞中的表达 总被引：1，自引：0，他引：1
目的:构建抑癌基因PTEN的表达载体并在LOVO细胞中表达。方法:从人外周血中提取总RNA,通过RT-PCR扩增出PTEN基因片段,将PTEN基因连入pLenti6/V5载体。测序鉴定后,经脂质体转染入LOVO细胞,对细胞株进行RT-PCR和Western blot检测。结果:DNA测序证实pLenti6/V5-PTEN表达载体为阳性克隆;该表达载体转染LOVO细胞后上调了PTEN mRNA和蛋白的表达。结论:成功构建了pLen-ti6/V5-PTEN表达载体,为进一步研究PTEN在LOVO细胞中的功能奠定了基础。 相似文献
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目的:探测细胞氧化低密度脂蛋白(LDL)过程中人α-防御素-1(HNP-1)基因的表达水平并构建人HNP-1的克隆载体。方法:提取人单核细胞系(THP-1)的总RNA,逆转录聚合酶链反应(RT-PCR)法获得cDNA,采用pBS-T克隆HNP-1。结果:用RT-PCR在THP-1细胞总RNA中扩增出一条285 bp的DNA片段,与HNP-1 cD-NA片段大小一致。重组质粒经PCR和测序鉴定获HNP-1基因克隆。另外,LDL可诱导THP-1细胞HNP-1的mR-NA表达增加。结论:HNP-1可能参与LDL的细胞氧化过程,成功构建HNP-1克隆载体将为进一步亚克隆到原核及真核的表达载体提供了基础。 相似文献
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Infusion of nonmyeloablative bone marrow alleviates acute rejection reaction in liver allotransplantation 总被引：1，自引：0，他引：1
Objective: To study the effect and implication of nonmyeloablative donor specific bone marrow (DSBM) infusion on the immunoreaction of liver allotransplantation. Methods: Orthotopic liver transplantation model was used in this study. Groups were set as follows: Group Ⅰ, syngeneic control (Wistar-to-Wistar); Group Ⅱ, acute rejection (SD-to-Wistar); Group Ⅲ, acute rejection treated with cyclosporine A (CsA) by intramuscular injection (SD-to-Wistar CsA); Group Ⅳ, bone marrow infusion at 7 d pretransplantation followed by short-term CsA treatment (SD-to-Wistar DSBM); Another group of short-term CsA treatment preoperatively without bone marrow infusion was also set as control. General characteristics and survival time were observed.Histological grades of rejection were determined by pathological examination. IL-2 and IFN-γ level in peripheral blood and donor liver were detected respectively by Enzyme-Linked Immuno-Sorbent Assay (ELISA) and Western blot. Chimerism of donor cells was measured by PCR for a male-specific marker (Y-chromosome-specific sequence, Sry). Results: No signs of rejection were found in Group Ⅰ. Acute rejection occurred in both Group Ⅱ and the short-term CsA treated group. All the recipients died at (9～15)d posttransplantation with a median survival time of (10.7±0.5) d and (11.2±2.4) d, respectively. Only mild rejection could be seen in Group Ⅲ. In Group Ⅳ, 4 out of 6 recipients had long-term survival (＞100 d), the histological grade of rejection was significantly lower than that of Group Ⅱ, so did the expression level of IL-2 and IFN-γ in both peripheral blood and grafted liver.Y-chromosome-specific sequence (Sry) of male SD rats could be detected in the bone marrow, spleen and thymus of female recipients at 15 d after bone marrow infusion. Conclusion: Mild preconditioning nonmyeloablative donor specific bone marrow infusion can enhance chimerism formation in recipients, alleviate the rejection of liver allotransplantation and prolong survival of liver allotransplantation. 相似文献
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It has been proposed that activation of Toll-like receptors (TLRs) plays crucial roles in the polarization of adaptive immune responses. A synthetic Toll-like receptor 2 (TLR2) ligand, Pam3CSK4, has been reported to modulate the balance of Thl/Tn2 responses. We evaluated the modulation effect of Pam3CSK4 on allergic immune response in a mouse rhinitis model sensitized to house dust mite allergen (HDM). Mice were sensitized and challenged with Dermatophagoides farinae allergen (Der f), and then the allergic mice were treated by Pam3CSK4. Nasal allergic symptoms and eosinophils were scored. Der f-specific cytokine responses were examined in the splenocytes and bronchoalveolar lavage fluid (BALF). Serum level of total IgE was also detected. After establishing a mouse allergic rhinitis model with HDM, we have showed that Pam3CSK4 treatment not only ameliorated the nasal allergic symptoms remarkably but also decreased the eosinophils and total inflammation cells in BALF significantly. Analysis of cytokine profile found that' IFN-γ released from either BALF or stimulated splenocytes increased markedly in Pam3CSK4-treated mice, while IL-13 decreased significantly. Moreover, serum level of total IgE was significantly lower in Pam3CSK4-treated mice than in the untreated. Thus, in an allergic rhinitis mouse model developed with HDM, Pam3CSK4 was shown to exhibit an antiallergic effect, indicating its potential application in allergic diseases. 相似文献