REGULATION OF GENE EXPRESSION IN HUMAN T CELL BY INTERLEUKIN 12, 1, 18 AND TNF
REGULATION OF GENE EXPRESSION IN HUMAN T CELL BY INTERLEUKIN 12, 1, 18 AND TNF
批准号:
6101213
负责人:
R. P DONNELLY
金额:
$0.0万
依托单位:
--
依托单位国家:
美国
项目类别:
财政年份:
--
资助国家:
美国
项目状态:
未结题
起止时间:
至
中文摘要
已知IL-12可上调血管内皮细胞合成干扰素-γ
活化的T细胞;然而,IL-12对其他T细胞表达的影响
细胞因子的定义不那么明确。在这个项目中,我们正在研究
IL-12对多种细胞因子产生的影响,包括干扰素-γ,
IL-2、IL-4、IL-10和IL-13,由纯化的正常人CD3 T细胞产生。
尽管静息T细胞在很大程度上对IL-12没有反应,
抗CD3激活的T细胞是高度IL-12反应的AS
IL-12诱导STAT4介导的DNA结合的能力
活动。我们已经发现,纯化的人类T淋巴细胞的激活
固定化抗CD3单抗诱导肿瘤坏死因子-α和白介素2的快速表达
干扰素-g、IL-10和IL-10的mRNA水平逐渐升高
IL-13。用核糖核酸酶测定mRNA表达水平的变化
使用便于分析的引物模板集进行保护分析
多个细胞因子基因同时存在。我们发现IL-12
显著上调干扰素-g和白介素10的表达,下调
IL-2的产生。IL-12对IL-2产生的抑制作用
直接与IL-10的产生增加有关。此外,中和
IL-10活性与抗IL-10抗体联合应用可使IL-2产生正常化
在IL-12处理的T细胞中显示出抑制作用
IL-12是由IL-10介导的。因此,我们发现IL-12同时
上调干扰素-g和白介素10的产生,并通过直接
IL-10依赖的途径,反馈抑制IL-2的产生。事实是
IL-12对T细胞干扰素-γ和IL-2合成的不同调节作用
细胞研究表明IL-12并不能整体增强ALL的表达
Th1型淋巴因子。此外,IL-12上调的能力
IL-10的产生提供了一种限制IL-2依赖的机制
活化T细胞的克隆性扩增及一种新的细胞因子
IL-12可诱导的调控途径。在未来的研究中,我们
计划进一步确定IL-12对其他物质生产的影响
细胞因子,特别是Th2型细胞因子IL-4和IL-13通过
激活T细胞,并比较IL-12与其他药物的作用
巨噬细胞衍生的免疫调节细胞因子,尤其是IL-1β
和肿瘤坏死因子-α。我们还将进一步探讨功能后果
IL-12处理的T细胞产生IL-10的增加。结果是
这些研究将扩大我们目前对
IL-12对免疫效应细胞的影响。此信息可能在以下方面有用
在临床试验中解释IL-12诱导的任何治疗活性
目前正在进行中。它还可以提供对生理学的洞察
与大剂量IL-12治疗相关的某些毒性的基础。
此外,还分析了IL-12对细胞因子表达的影响。
体外实验可能有助于IL-12替代标志物的鉴定
生物活性可用于监测重组蛋白的效力
IL-12在人类受体中的表达。
英文摘要
IL-12 is known to up-regulate synthesis of interferon-gamma (IFN-g) in
activated T cells; however, the effects of IL-12 on expression of other
cytokines are less well defined. In this project, we are examining the
effects of IL-12 on production of multiple cytokines, including IFN-g,
IL-2, IL-4, IL-10 and IL-13, by purified, normal, human CD3+ T cells.
Although resting T cells are largely IL-12-nonresponsive,
anti-CD3-activated T cell blasts are highly IL-12-responsive as
demonstrated by the ability of IL-12 to induce STAT4-mediated DNA-binding
activity. We have found that activation of purified human T lymphocytes
on immobilized anti-CD3 mAb induces rapid expression of TNF-a and IL-2
mRNA, and more gradual increases in mRNA levels for IFN-g, IL-10 and
IL-13. Changes in mRNA expression levels were measured by RNase
protection assay using primer template sets that facilitate analysis of
multiple cytokine genes simultaneously. We have found that IL-12
markedly up-regulates expression of IFN-g and IL-10, and down-regulates
production of IL-2. Inhibition of IL-2 production by IL-12 correlates
directly with increased production of IL-10. Moreover, neutralization
of IL-10 activity with anti-IL-10 antibodies normalizes IL-2 production
in IL-12-treated T cells demonstrating that the inhibitory effects of
IL-12 are IL-10-mediated. Thus, we have found that IL-12 simultaneously
up-regulates production of IFN-g and IL-10, and, by a direct
IL-10-dependent pathway, feedback inhibits production of IL-2. The fact
that IL-12 differentially regulates synthesis of IFN-g and IL-2 in T
cells demonstrates that IL-12 does not globally enhance expression of all
Th1-type lymphokines. Furthermore, the ability of IL-12 to up-regulate
production of IL-10 provides a mechanism for limiting the IL-2-dependent
clonal expansion of activated T cells, and defines a novel cytokine
regulatory pathway that is inducible by IL-12. In future studies, we
plan to further define the effects of IL-12 on production of other
cytokines, particularly the Th2-type cytokines IL-4 and IL-13 by
activated T cells, and to compare the effects of IL-12 with that of other
macrophage-derived immunoregulatory cytokines, most notably IL-1 beta
and TNF-alpha. We will also further explore the functional consequences
of increased IL-10 production in IL-12-treated T cells. The results of
these studies will broaden our current understanding of the actions of
IL-12 on immune effector cells. This information may be useful in
interpreting any therapeutic activity induced by IL-12 in clinical trials
that are now underway. It may also provide insight to the physiological
basis for certain toxicities associated with high dose IL-12 therapy.
Furthermore, analysis of the effects of IL-12 on cytokine expression in
vitro may facilitate identification of surrogate markers of IL-12
bioactivity that could be useful in monitoring the potency of recombinant
IL-12 in human recipients.
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会议论文
REGULATION OF CYTOKINE GENE EXPRESSION IN HUMAN T CELLS BY IL-4 AND IL12
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批准号:6293751
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:R. P DONNELLY
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依托单位:--
REGULATION OF HUMAN T CELL FUNCTIONS BY INTERLEUKIN 12 (IL-12)
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批准号:2456633
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:R. P DONNELLY
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依托单位:--
REGULATION OF GENE EXPRESSION BY INTERLEUKIN10 IN ENDOTOXIN-STIMULATED HUMAN MONO
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批准号:6293756
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:R. P DONNELLY
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依托单位:--
Regulation of Monocyte Gene Expression
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批准号:6839785
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:R. P DONNELLY
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依托单位:--
Characterization of Novel Interleukin 10 Related Genes
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批准号:6545298
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:R. P DONNELLY
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依托单位:--
Recombinant Human Interleukin 12 and Cytokine Expression
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批准号:6545297
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:R. P DONNELLY
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依托单位:--
REGULATION OF CYTOKINE PRODUCTION BY IL-10 IN ENDOTOXIN-STIMULATED MONOCYTES
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批准号:6101222
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:R. P DONNELLY
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依托单位:--
IDENTIFICATION & CHARACTERIZATION OF IL-10 RELATED GENES
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批准号:6436327
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:R. P DONNELLY
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依托单位:--
Characterization of the Effects of Recombinant Human Int
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批准号:6679779
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:R. P DONNELLY
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依托单位:--
REGULATION OF CYTOKINE PRODUCTION BY IL-10 IN ENDOTOXIN-STIMULATED MONOCYTES
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批准号:2568967
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:R. P DONNELLY
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依托单位:--
EFFECTS OF RECOMBINANT IL12 ON CYTOKINE GENE EXPESSION
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批准号:6436317
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:R. P DONNELLY
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依托单位:--
REGULATION OF GENE EXPRESSION IN HUMAN T CELL BY INTERLEUKIN 12, 1, 18 AND TNF
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批准号:6161275
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:R. P DONNELLY
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依托单位:--
REGULATION OF CYTOKINE PRODUCTION BY IL-10 IN ENDOTOXIN-STIMULATED MONOCYTES
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批准号:6161284
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:R. P DONNELLY
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依托单位:--
Identification and Characterization of Novel Interleukin
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批准号:6679780
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:R. P DONNELLY
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依托单位:--
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