Smith J K, Chi D S, Krishnaswamy G, Srikanth S, Reynolds S, Berk S L
Department of Internal Medicine, East Tennessee State University, Johnson City 37614-0622, USA.
Arch Immunol Ther Exp (Warsz). 1996;44(2-3):83-8.
/We have studied the effect of interferon alpha (IFN-alpha) on MHC class II expression by human buccal epithelial cells (BEC), and mRNA expression by BEC and mucosal-associated mononuclear cells (MAMC). In 6 experiments, freshly collected BEC were suspended at a concentration of 1.0 x 10(5)/ml in RPMI 1640 and incubated in the presence of 0-10,000 IU/ml of human lymphoblastoid IFN-alpha (HuIFN-alpha). Zero and six hour samples were analyzed by single color flow cytometry using FITC-labeled murine IgG1 monoclonal antibody to HLA-DR. Preparations were also analyzed for expression of cytokine transcripts (IL-2, IL-4, IL-5, IL-6, IL-8, IFN-gamma, GM-CSF) by reverse transcriptase polymerase chain reaction (RT-PCR). Increasing concentrations of IFN-alpha resulted in proportionate increases in the percentage of HLA-DR + BEC (r = 0.7897, p = 0.0627) and in the percentage of HLA-DR+ staining at higher intensities (10(1) to 10(2) log fluorescence intensity) (LFI) (r = 0.4010, p = 0.0424). The percentage of HLA-DR + BEC rose from a mean of 1.5% with no IFN-alpha to 7% with 10,000 IU/ml IFN-alpha (p < 0.05). The percentage of HLA-DR + BEC staining at 10(1) to 10(2) LFI rose from a mean of 8.3% with no added IFN-alpha to 19.2% with 10,000 IU/ml IFN-alpha (p < 0.05). Unstimulated BEC constitutively expressed IL-8 and GM-CSF. IFN-alpha stimulated preparations also expressed IFN-gamma, possibly due to the presence of MAMC, which comprised 2-9% of the total cell population. These data indicated that HuIFN-alpha upregulates MHC class II expression by human BEC, possibly by enhancing IFN-gamma production by MAMC present in the culture preparations.
我们研究了α干扰素(IFN-α)对人颊黏膜上皮细胞(BEC)主要组织相容性复合体II类(MHC II)表达以及对BEC和黏膜相关单核细胞(MAMC)mRNA表达的影响。在6项实验中,将新鲜采集的BEC以1.0×10⁵/ml的浓度悬浮于RPMI 1640中,并在0 - 10,000 IU/ml的人淋巴母细胞样IFN-α(HuIFN-α)存在的情况下进行培养。使用异硫氰酸荧光素(FITC)标记的抗HLA - DR鼠IgG1单克隆抗体,通过单色流式细胞术对0小时和6小时的样本进行分析。还通过逆转录聚合酶链反应(RT-PCR)分析细胞因子转录本(IL-2、IL-4、IL-5、IL-6、IL-8、IFN-γ、GM-CSF)的表达情况。IFN-α浓度的增加导致HLA - DR⁺ BEC百分比成比例增加(r = 0.7897,p = 0.0627),并且在较高强度(10¹至10²对数荧光强度)(LFI)下HLA - DR⁺染色百分比也增加(r = 0.4010,p = 0.0424)。HLA - DR⁺ BEC的百分比从无IFN-α时的平均1.5%上升至10,000 IU/ml IFN-α时的7%(p < 0.05)。在10¹至10² LFI下HLA - DR⁺ BEC染色百分比从无添加IFN-α时的平均8.3%上升至10,000 IU/ml IFN-α时的19.2%(p < 0.05)。未受刺激的BEC组成性表达IL-8和GM-CSF。IFN-α刺激的样本还表达IFN-γ,这可能是由于存在占总细胞群体2 - 9%的MAMC。这些数据表明,HuIFN-α可能通过增强培养样本中存在的MAMC产生IFN-γ,从而上调人BEC的MHC II表达。