Richards D, Chapman M D, Sasama J, Lee T H, Kemeny D M
Department of Allergy and Respiratory Medicine, United Medical School of Guy's Hospital, London, UK.
Immunology. 1997 Jul;91(3):331-9. doi: 10.1046/j.1365-2567.1997.00274.x.
This study addresses the question of whether human peripheral CD4+ CD45RA+ T cells possess antigen-specific immune memory. CD4+ CD45RA+ T cells were isolated by a combination of positive and negative selection. Putative CD4+ CD45RA+ cells expressed CD45RA (98.9%) and contained < 0.1% CD4+ CD45RO+ and < 0.5% CD4+ CD45RA+ CD45RO+ cells. Putative CD45RO+ cells expressed CD45RO (90%) and contained 9% CD45RA+ CD45RO+ and < 0.1% CD4+ CD45RA+ cells. The responder frequency of Dermatophagoides pteronyssinus-stimulated CD4+ CD45RA+ and CD4+ CD45RO+ T cells was determined in two atopic donors and found to be 1:11,314 and 1:8031 for CD4+ CD45RA+ and 1:1463 and 1:1408 for CD4+ CD45RO+ T cells. The responder frequencies of CD4+ CD45RA+ and CD4+ CD45RO+ T cells from two non-atopic, but exposed, donors were 1:78031 and 1:176,903 for CD4+ CD45RA+ and 1:9136 and 1:13,136 for CD4+ CD45RO+ T cells. T cells specific for D. pteronyssinus were cloned at limiting dilution following 10 days of bulk culture with D. pteronyssinus antigen. Sixty-eight clones were obtained from CD4+ CD45RO+ and 24 from CD4+ CD45RA+ T cells. All clones were CD3+ CD4+ CD45RO+ and proliferated in response to D. pteronyssinus antigens. Of 40 clones tested, none responded to Tubercule bacillus purified protein derivative (PPD). No difference was seen in the pattern of interleukin-4 (IL-4) or interferon-gamma (IFN-gamma) producing clones derived from CD4+ CD45RA+ and CD4+ CD45RO+ precursors, although freshly isolated and polyclonally activated CD4+ CD45RA+ T cells produced 20-30-fold lower levels of IL-4 and IFN-gamma than their CD4+ CD45RO+ counterparts. Sixty per cent of the clones used the same pool of V beta genes. These data support the hypothesis that immune memory resides in CD4+ CD45RA+ as well as CD4+ CD45RO+ T cells during the chronic immune response to inhaled antigen.
本研究探讨了人类外周CD4+ CD45RA+ T细胞是否具有抗原特异性免疫记忆这一问题。通过阳性和阴性选择相结合的方法分离出CD4+ CD45RA+ T细胞。假定的CD4+ CD45RA+细胞表达CD45RA(98.9%),且含有<0.1%的CD4+ CD45RO+细胞以及<0.5%的CD4+ CD45RA+ CD45RO+细胞。假定的CD45RO+细胞表达CD45RO(90%),且含有9%的CD45RA+ CD45RO+细胞以及<0.1%的CD4+ CD45RA+细胞。在两名特应性供体中测定了尘螨刺激的CD4+ CD45RA+和CD4+ CD45RO+ T细胞的应答频率,发现CD4+ CD45RA+ T细胞的应答频率分别为1:11,314和1:8031,CD4+ CD45RO+ T细胞的应答频率分别为1:1463和1:1408。来自两名非特应性但暴露的供体的CD4+ CD45RA+和CD4+ CD45RO+ T细胞的应答频率,CD4+ CD45RA+ T细胞分别为1:78031和1:176,903,CD4+ CD45RO+ T细胞分别为1:9136和1:13,136。在用尘螨抗原进行10天的大量培养后,通过有限稀释法克隆针对尘螨的T细胞。从CD4+ CD45RO+ T细胞中获得了68个克隆,从CD4+ CD45RA+ T细胞中获得了24个克隆。所有克隆均为CD3+ CD4+ CD45RO+,并对尘螨抗原产生增殖反应。在测试的40个克隆中,没有一个对结核杆菌纯化蛋白衍生物(PPD)产生反应。源自CD4+ CD45RA+和CD4+ CD45RO+前体的产生白细胞介素-4(IL-4)或干扰素-γ(IFN-γ)的克隆模式没有差异,尽管新鲜分离和多克隆激活的CD4+ CD45RA+ T细胞产生的IL-4和IFN-γ水平比其CD4+ CD45RO+对应细胞低20 - 30倍。60%的克隆使用相同的Vβ基因库。这些数据支持了这样一种假设,即在对吸入抗原的慢性免疫反应过程中,免疫记忆存在于CD4+ CD45RA+以及CD4+ CD45RO+ T细胞中。