CYTOKINE GENE-EXPRESSION IN MICE UNDERGOING CHRONIC GRAFT-VERSUS-HOST DISEASE
CYTOKINE GENE-EXPRESSION IN MICE UNDERGOING CHRONIC GRAFT-VERSUS-HOST DISEASE
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DOI：
10.1016/0161-5890(93)90078-p
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发表时间：
1993-05-01
影响因子：
3.6
通讯作者：
UMLAND, SP
中科院分区：
文献类型：
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作者：
GARLISI, CG;PENNLINE, KJ;UMLAND, SP
通过注射亲代DBA/2淋巴样细胞可以在B6 D2 F1小鼠中诱导慢性移植物抗宿主病（GVHD）。宿主MHC抗原对供体T细胞的刺激导致对宿主B细胞的刺激。人们对这种反应中产生的淋巴因子知之甚少。本研究旨在使用半定量聚合酶链反应（PCR）直接测量干扰素-γ（IFN-γ）、白细胞介素2（IL-2）、IL-4、IL-5和IL-10以及其他几种基因的mRNA水平。半定量PCR是可重复的，并且产生的信号取决于每个反应中特异性RNA或cDNA的量。在GVHD进展的早期（第一次注射亲本细胞后2天），IL-10 mRNA几乎没有增加，IL-4 mRNA略有增加，IL-2 mRNA急剧增加。体外培养24 h的GVH脾细胞培养上清中IL-2活性明显增强。在反应后期（第二次和最后一次注射亲本细胞后1周），IL-4 mRNA水平升高，因为它们更早，而IL- 10 mRNA水平显著增加。IL-2 mRNA水平在经历GVHD的小鼠中与正常小鼠相比没有差异。IFN-γ mRNA在早期和晚期都可检测到，尽管在正常小鼠和经历GVHD的小鼠中水平相似。在检查的两个时间点，IL-4均低于生物测定法的检测限，IFN-γ、IL-4、IL-5和IL-10均低于ELISA法的检测限。进一步的研究表明，在GVH小鼠中发现的升高的IL-4和IL-10 mRNA大多数由Thy1.2+ T细胞产生，少量来自B220+ B细胞。此外，在GVH小鼠中发现的可检测的IFN-γ mRNA也由Thy1.2+ T细胞产生，少量来自B220+ B细胞。
Chronic graft-versus-host disease (GVHD) can be induced in B6D2F1 mice by injection of parental DBA/2 lymphoid cells. Stimulation of donor T cells by host MHC antigens leads to the stimulation of host B cells. Little is known of the lymphokines produced during such a reaction. This study was designed to directly measure the levels of mRNA for interferon-gamma (IFN-gamma), interleukin 2 (IL-2), IL-4, IL-5, and IL-10, as well as several other genes, using semiquantitative polymerase chain reaction (PCR). Semiquantitative PCR was reproducible and signals generated were dependent on the amount of specific RNA or cDNA in each reaction. Early during the progression of GVHD (2 days after the first injection of parental cells) there was little increase in IL-10 mRNA, a slight increase in IL-4 mRNA, and a dramatic increase in IL-2 mRNA. In addition, IL-2 bioactivity was demonstrated in supernatants from GVH splenocytes cultured in vitro for 24 h. Later in the response (I week after the second and final injection of parental cells) IL-4 mRNA levels were elevated as they were earlier while IL- 10 mRNA levels were dramatically increased. IL-2 mRNA levels were no different in mice undergoing GVHD than in normal mice at this time. IFN-gamma mRNA was detectable both early and late, although at similar levels in normal mice and mice undergoing GVHD. At both times examined, IL-4 was below the limits of detection by bioassay and IFN-gamma, IL-4, IL-5 and IL-10 were below the limits of detection by ELISA. Further studies showed that a majority of the IL-4 and IL-10 mRNA found elevated in GVH mice were produced by Thy1.2+ T cells, with small amounts from B220+ B cells. In addition, the detectable IFN-gamma mRNA found in GVH mice at this later time also was produced by Thy1.2+ T cells, with small amounts from B220+ B cells.