共查询到20条相似文献，搜索用时 15 毫秒
1.
Human GM-CSF: molecular cloning of the complementary DNA and purification of the natural and recombinant proteins 总被引：185，自引：0，他引：185
G G Wong J S Witek P A Temple K M Wilkens A C Leary D P Luxenberg S S Jones E L Brown R M Kay E C Orr 《Science (New York, N.Y.)》1985,228(4701):810-815
Clones of complementary DNA encoding the human lymphokine known as granulocyte-macrophage colony-stimulating factor (GM-CSF) were isolated by means of a mammalian cell (monkey COS cell) expression screening system. One of these clones was used to produce recombinant GM-CSF in mammalian cells. The recombinant hematopoietin was similar to the natural product that was purified to apparent homogeneity from medium conditioned by a human T-cell line. The human T-cell GM-CSF was found to be 60 percent homologous with the GM-CSF recently cloned from murine lung messenger RNA. 相似文献
2.
Biological activity of recombinant human interleukin-2 produced in Escherichia coli 总被引：72，自引：0，他引：72
S A Rosenberg E A Grimm M McGrogan M Doyle E Kawasaki K Koths D F Mark 《Science (New York, N.Y.)》1984,223(4643):1412-1414
The gene for interleukin-2 was isolated from the Jurkat cell line and from normal peripheral blood lymphocytes and, when inserted in Escherichia coli, was expressed at high concentrations. This interleukin-2 was purified to apparent homogeneity and tested for biological activity in a variety of assays in vitro and in vivo. The recombinant lymphokine supports the growth of murine and human interleukin-2 dependent cell lines, enhances the generation of murine and human cytolytic cells in vitro, and generates lymphokine activated killer cells from murine and human lymphocytes. It has a serum half-life of 2 to 3 minutes in the mouse and significantly enhances the generation of cytolytic cells in vivo after alloimmunization. No functional differences between native and the recombinant interleukin-2 molecules have been detected. 相似文献
3.
Serological visualization of interleukin 2 总被引：5，自引：0，他引：5
Interleukin 2, a lymphokine that acts as a second signal of cellular immune response by way of its action as a T-cell growth factor, was morphologically identified by immunoperoxidase staining. With the use of a monoclonal antibody to interleukin 2 and several complex-forming antisera, the lymphokine was readily distinguished in cytocentrifuge preparations of peripheral blood leukocytes stimulated with a T-cell mitogen. When preparations of cloned interleukin 2 producer and responder cells were stained by the same procedures, discrete patterns of both responder and producer cell phenotypes were revealed. Interleukin 2 producer T cells exhibited a characteristic intense, ringlike cytoplasmic staining, whereas the responder cells (as exemplified by interleukin 2-dependent cell lines) exhibited a less intensive, spotlike membrane staining. In addition, intense membrane localization of interleukin 2, reminiscent of potential capping phenomena, could be observed in stained preparations of cloned responder cells. 相似文献
4.
Prostacyclin synthesis induced in vascular cells by interleukin-1 总被引：26，自引：0，他引：26
Supernatants from cultures of human monocytes that had been stimulated with endotoxin or silica induced the synthesis of prostacyclin in endothelial and smooth muscle cells. The lymphokine mediating these effects on the cells of the blood vessel wall was identified as interleukin-1; interferons and interleukin-2 were inactive. Interleukin-1-induced prostacyclin synthesis represents a new aspect of the interaction between the immune system (as well as other tissues) and the vessel wall and may serve as a basis for the development of new strategies in antithrombotic therapy. 相似文献
5.
Mechanism of interleukin-2 signaling: mediation of different outcomes by a single receptor and transduction pathway 总被引：18，自引：0，他引：18
The T cell lymphokine, interleukin-2 (IL-2), plays a pivotal role in an immune response by stimulating antigen-activated B lymphocytes to progress through the cell cycle and to differentiate into antibody-secreting cells. An IL-2 inducible B lymphoma line, in which the growth and differentiation responses are uncoupled, provides a model system for dissecting the signaling mechanisms operating in each response. This system was used to show that both signals are initiated by IL-2 binding to a single, unifunctional receptor complex. Moreover, both signals are transduced by a pathway that does not involve any known second messenger system and that can be blocked by a second T cell lymphokine, interleukin 4. These findings suggest that the pleiotrophic effects of IL-2 are determined by different translations of the signal in the nucleus. 相似文献
6.
Induction of interleukin 2 messenger RNA inhibited by cyclosporin A 总被引：48，自引：0，他引：48
J F Elliott Y Lin S B Mizel R C Bleackley D G Harnish V Paetkau 《Science (New York, N.Y.)》1984,226(4681):1439-1441
Cyclosporin A blocked production of the lymphokine interleukin 2 by activated T lymphocytes. In a human and a murine cell line this inhibition reflected an absence of interleukin 2 messenger RNA. Under conditions in which these cells are normally stimulated to secrete high levels of interleukin 2, they failed to do so in the presence of cyclosporin A. In both cell lines this failure was accompanied by an absence of interleukin 2 messenger accumulation. 相似文献
7.
[目的]构建稳定表达犬瘟热病毒细胞受体———犬信号淋巴细胞激活因子(SLAM)的非洲绿猴肾细胞株(Vero)。[方法]采用RT-PCR方法从犬外周血淋巴细胞中扩增出SLAM基因,将其克隆到哺乳动物真核表达载体pcDNA3.1(+)中,构建重组质粒pcDNA3.1/SLAM。采用脂质体将pcDNA3.1/SLAM转染到Vero细胞中,利用G418加压筛选和纯化培养获得稳定表达SLAM的重组Vero细胞株。应用RT-PCR和间接免疫荧光试验检测SLAM的表达。[结果]重组蛋白SLAM在Vero细胞中获得表达,并且在不同代次的阳性细胞株中均能稳定表达目的蛋白。[结论]该研究建立了稳定表达犬SLAM的细胞株Vero/SLAM,为犬瘟热病毒的分离和生物学特性研究提供了平台。 相似文献
8.
猪粒细胞-巨噬细胞集落刺激因子(GM-CSF)基因的克隆和序列分析及其基因表达 总被引：1，自引：1，他引：0
采取三元杂交猪的外周血,提取单核细胞总RNA,采用RT-PCR克隆猪粒细胞-巨噬细胞集落刺激因子(GM-CSF)cDNA(GenBank登录号为DQ108393)。以pcDNA3.1(-)为质粒载体,构建猪GM-CSF基因真核表达质粒pGM-CSF,对克隆片段进行测序,并进行不同物种间同源性的比较。将重组质粒转染COS-7细胞,Western blotting鉴定表达蛋白的特异性。序列分析表明,GM-CSF的cDNA开放阅读框为435 bp,编码144个氨基酸和大部分信号肽,与用作PCR引物设计的参考序列比较,其核苷酸的同源性为97.5%,氨基酸的同源性为95.1%,同绵羊、人、牛、小鼠和犬的GM-CSF序列比较,其核苷酸的同源性分别为86.2%、80.5%、81.7%、69.7%和81.8%,氨基酸的同源性分别为78.5%、71.3%、70.3%、55.9%和71.5%。Western blotting证实目的基因可以在真核细胞中特异性的表达GM-CSF蛋白。以上结果表明,猪的GM-CSF基因被成功克隆,它存在着种属特异性。这为进一步研究该基因的生物学作用特别是利用GM-CSF增强DNA疫苗的免疫效果奠定了基础。 相似文献
9.
Human recombinant granulocyte-macrophage colony-stimulating factor: a multilineage hematopoietin 总被引：33，自引：0，他引：33
C A Sieff S G Emerson R E Donahue D G Nathan E A Wang G G Wong S C Clark 《Science (New York, N.Y.)》1985,230(4730):1171-1173
Human recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF) was tested for its ability to induce colony formation in human bone marrow that had been enriched for progenitor cells. In addition to its expected granulocyte-monocyte colony-stimulating activity, the recombinant GM-CSF had burst-promoting activity for erythroid burst-forming units and also stimulated colonies derived from multipotent (mixed) progenitors. In contrast, recombinant erythroid-potentiating activity did not stimulate erythroid progenitors. The experiments prove that human GM-CSF has multilineage colony-stimulating activity. 相似文献
10.
粒细胞-巨噬细胞集落刺激因子(GM-CSF)是机体免疫系统的重要细胞因子,具有生物佐剂作用,为研究GM-CSF的生物佐剂作用,本试验通过RT-PCR方法从小鼠脾脏细胞中扩增小鼠GM-CSF的cDNA,并将其插入到pcDNA3.1质粒中,构建成GM-CSF真核表达载体pcDNA3.1-GMCSF,并在真核细胞进行了瞬时表达。结果表明,本研究扩增的小鼠GM-CSF基因序列与GenBank序列完全一致,表达载体经脂质体介导转染HEK293T细胞,表达产物经western-blot检测,证明GM-CSF能够在HEK293T细胞中进行分泌表达。这为今后研究GM-CSF在动物疫苗,特别是DNA疫苗中的生物佐剂作用创造了必要的条件。 相似文献
11.
The structure, function, and expression of interleukin-2 receptors on normal and malignant lymphocytes 总被引：56，自引：0，他引：56
T A Waldmann 《Science (New York, N.Y.)》1986,232(4751):727-732
Antigen or mitogen-induced activation of resting T cells induces the synthesis of interleukin-2 (IL-2) as well as the expression of specific cell surface receptors for this lymphokine. Failure of the production of either IL-2 or its receptor results in a failure of the T-cell immune response. The receptor is composed of a 33,000-dalton (251-amino acid) peptide precursor that is post-translationally glycosylated into the mature 55,000-dalton form. In contrast to resting T cells, human T-cell lymphotrophic virus I (HTLV-I)-associated adult T-cell leukemia cells constitutively express large numbers of IL-2 receptors. Because IL-2 receptors are present on the malignant T cells but not on normal resting cells, clinical trials have been initiated in which patients with adult T-cell leukemia are treated with a monoclonal antibody that binds to the IL-2 receptor. 相似文献
12.
Human malignant cells secrete low molecular size proteins that attract peripheral blood monocytes and may be responsible for the accumulation of tumor-associated macrophages observed in vivo. Similar chemotactic proteins are secreted by cultured vascular smooth muscle cells. The predominant monocyte chemoattractants produced by tumor cells of differing origin were demonstrated to be related to smooth muscle cell-derived chemotactic factor. Thus, a single class of chemotactic proteins is produced by different cell types, which suggests a common mechanism for the recruitment of monocytes and macrophages. These results are significant in view of the potential of macrophages to affect tumor growth. 相似文献
13.
Immunoregulatory lymphokines of T hybridomas from AIDS patients: constitutive and inducible suppressor factors 总被引：11，自引：0，他引：11
Supernatants derived from peripheral blood mononuclear cell cultures of certain patients with the acquired immunodeficiency syndrome (AIDS) or its prodromes have the capacity to block T cell-dependent immune reactivity in vitro. T cells derived from a patient positive for antibody to the lymphadenopathy associated virus ( LAV ), and elaborating high titers of these soluble suppressor factors, were fused to a mutagenized clone of the human T lymphoblastoid cell line KE37 . Molecules capable of profoundly depressing T cell-dependent polyclonal antibody production and DNA synthetic responses, either directly or after incubation with normal adherent cells, were isolated from stable hybrid clones. 相似文献
14.
Identification of a T helper cell-derived lymphokine that activates resting T lymphocytes 总被引：3，自引：0，他引：3
A novel lymphokine with apparent molecular size of 10 to 12 kilodaltons is secreted from helper T cell clones within hours after cross-linking their T cell antigen-MHC (major histocompatibility complex) receptors (T3-Ti). This lymphokine, termed interleukin-4A (IL-4A), stimulates resting lymphocytes by binding to a surface component (or components) of the alternative T11 pathway and subsequently by inducing interleukin-2 (IL-2) receptors. The activation process is neither dependent on antigen specificities of the recruited population or the presence of macrophages. It appears, therefore, that IL-4A is a mediator involved in amplifying the T cell immune response. 相似文献
15.
Beatty GL Chiorean EG Fishman MP Saboury B Teitelbaum UR Sun W Huhn RD Song W Li D Sharp LL Torigian DA O'Dwyer PJ Vonderheide RH 《Science (New York, N.Y.)》2011,331(6024):1612-1616
Immunosuppressive tumor microenvironments can restrain antitumor immunity, particularly in pancreatic ductal adenocarcinoma (PDA). Because CD40 activation can reverse immune suppression and drive antitumor T cell responses, we tested the combination of an agonist CD40 antibody with gemcitabine chemotherapy in a small cohort of patients with surgically incurable PDA and observed tumor regressions in some patients. We reproduced this treatment effect in a genetically engineered mouse model of PDA and found unexpectedly that tumor regression required macrophages but not T cells or gemcitabine. CD40-activated macrophages rapidly infiltrated tumors, became tumoricidal, and facilitated the depletion of tumor stroma. Thus, cancer immune surveillance does not necessarily depend on therapy-induced T cells; rather, our findings demonstrate a CD40-dependent mechanism for targeting tumor stroma in the treatment of cancer. 相似文献
16.
为探明原核表达的重组猪IL-10对仔猪的免疫原性,根据GenBank公布的猪IL-10基因序列(登录号:L20001)设计并合成了扩增猪IL-10基因的引物。利用ConA刺激猪外周血淋巴细胞,通过RT-PCR扩增出pmIL-10完整基因及其成熟蛋白基因片段(pmIL-10),分别插入质粒pVAX1和pET32α(+),对猪IL-10基因进行原核与真核表达。利用纯化的重组蛋白免疫仔猪,制备抗血清,利用间接ELISA检测血清效价。将pVAX-pIL-10转染COS-7细胞,利用RT-PCR检测pmIL-10的转录,以间接免疫荧光(IFA)鉴定其与原核表达产物制备抗体之间的反应。结果显示:该试验成功获得了猪IL-10基因,重组菌pET-pmIL-10/Rosetta经IPTG诱导后,猪IL-10获得表达,目的蛋白表达量以包涵体形式存在,占菌体总蛋白的39.7%;以白油佐剂重组蛋白免疫仔猪可产生高水平的抗猪IL-10的抗体,经ELISA检测,P/N值可达到8,OD630吸光值可达1.875,该抗体与转染pVAX-pmIL-10的COS-7细胞发生明显的荧光反应,说明原核表达的重组猪IL-10免疫产生的抗体具有生物学活性,为日后利用猪IL-10抗体消除病毒感染中产生的内源性IL-10研究奠定了基础。 相似文献
17.
Epitopes recognized by human T cells map within the conserved part of the GP190 of P. falciparum 总被引：10，自引：0，他引：10
A Crisanti H M Müller C Hilbich F Sinigaglia H Matile M McKay J Scaife K Beyreuther H Bujard 《Science (New York, N.Y.)》1988,240(4857):1324-1326
In a study aimed at developing a vaccine against the asexual blood stages of Plasmodium falciparum, two T cell epitopes were identified within a nonpolymorphic region of gp190 of Plasmodium falciparum merozoites. The two epitopes, which were revealed by deletion analysis, stimulated human T cell clones. Peptides containing sequences of the epitopes stimulated the cloned T cells and peripheral blood mononuclear cells (PBMC) from malaria-infected individuals. Moreover, the T cell clones responded to 11 different Plasmodium falciparum isolates in culture, showing that the epitopes are recognized in native parasites. 相似文献
18.
Purified human granulocyte-macrophage colony-stimulating factor: direct action on neutrophils 总被引：57，自引：0，他引：57
J C Gasson R H Weisbart S E Kaufman S C Clark R M Hewick G G Wong D W Golde 《Science (New York, N.Y.)》1984,226(4680):1339-1342
Neutrophil migration inhibition factor from T lymphocytes (NIF-T) is a lymphokine that acts to localize granulocytes. Medium conditioned by the Mo human T-lymphoblast cell line was used to purify NIF-T, a glycoprotein with a molecular weight of 22,000. The NIF-T was found to potently stimulate the growth of granulocyte and macrophage colonies from human bone marrow and colony formation by the KG-1 myeloid leukemia cell line. Thus a human lymphokine (NIF-T) that modulates the activities of mature neutrophilic granulocytes is also a colony-stimulating factor acting on precursors to induce growth and differentiation of new effector cells. 相似文献
19.
20.
Cytokines alter production of HIV-1 from primary mononuclear phagocytes 总被引：58，自引：0，他引：58
Y Koyanagi W A O'Brien J Q Zhao D W Golde J C Gasson I S Chen 《Science (New York, N.Y.)》1988,241(4873):1673-1675
Some strains of human immunodeficiency virus type 1 (HIV-1) can infect primary monocytes and monocyte-derived macrophages in vitro. In this report, the effect of cytokines on the production of one of these strains that shows a tropism for mononuclear phagocytes, designated HIV-1JR-FL, was studied. Primary peripheral blood mononuclear phagocytes infected with HIV-1JR-FL were treated with the hematopoietic factors: granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3), macrophage colony-stimulating factor (M-CSF), and gamma-interferon (gamma-IFN). The M-CSF, GM-CSF, IL-3, and gamma-IFN were able to alter HIV-1 production under different conditions. 相似文献