Ⅱ类抗原反式激活因子与HLADR抗原的关系及其意义.docVIP

　　Ⅱ类抗原反式激活因子与HLADR抗原的关系及其意义 【摘要】 本 研究 探讨Ⅱ类抗原反式激活因子（CIITA）和人类白细胞抗原（HLADR）表达时相的关系和差异，及STAT1α反义寡核苷酸（STAT1α AS）对CIITA和HLADR的抑制作用。分离健康志愿者外周血T淋巴细胞，给予不同剂量干扰素γ（IFNγ）后，用RTPCR法检测CIITA mRNA，NC) heparinized peripheral blood of normal donors using FicollHypaque densitygradient centrifugation and ents. To obtain a monocyteenriched cell population, PBMNC edium (Gibco) containing 100 U/ml penicillin, 100 U/ml streptomycin and 10% heatinactivated fetal calf serum (FCS). Nonadherent cells oved by ed (37℃) culture medium. Then adherent cells edium. CD3positive cells accounted for 70% of the nonadherent cell population. Interferonγ (IFNγ, Shanghai Clone HighTech pany, China) l) cultures at different final concentration of 50 U/ml to 1200 U/ml for induction of HLADR antigen at 37℃, 5% CO2. T cells RNA, and HLADR protEin ent. Samples erase chain reaction (RTPCR) Total RNA anufacturer. Singlestranded cDNA 2 μg of total RNA in a 40 μl reaction mixture containing 0.2 μg random primer and 300 U reverse transcriptase (Promega, Madison, edium oved and replaced l IFNγ. Then the experiment edium only contained 1000 U/ml IFNγ and RPMI 1640, not incubated RNA by RTPCR at 14 hours, or cells ents es ean±SD of four independent experiments. Statistical analysis of the results ed by oneultiple range method. Plt;0.05 HⅠ site. The PCR products of CIITA gene HⅠ, and tents ents after digestion arker. Lane 1: bands after enzymolysed by BamH1 (308 bp and 163 bp). Lane 2： PCR products (471 bp) Expression of CIITA mRNA in T cells Expression of CIITA l and 1000 U/ml. : marker. CIITA gene products edependent manner. IFNγ induced HLADR protein ulation ulation (Figure 3). CIITA expression l IFNγ treatment at various time points (hour). Inhibition of CIITA and HLADR expression by antisense oligonucleotides against STAT1α There RNA betol/L. The expression of CIITA mRNA ol/L (Plt;0.01), but there ol/L AS group and 20 μmol/L AS group (Pgt;0.05). The expression level of CIITA mRNA