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用固相放射免疫试验从250名志愿者中筛选出4名高抗-HBs者作为试验对象。将其外周血离心,提取单个核细胞(PBM),6,000万个PBM细胞于37℃放置1小时,收集非粘附性细胞于低温保存。粘附性细胞用高度提纯的HBsAg(2μg/ml)于37℃处理90分钟。从非粘附性细胞中去除抑制性T细胞后,轻轻地倒在经抗原处理过的单层粘附性细胞上,加美洲商陆有丝分裂原(PWM,1:200稀释)或HBsAg(2μg/ml)和PWM(1:200稀释),培养5天。4名供血者中,仅1人对HBsAg刺激的应答较弱;其余3人应答均较强。
Four high anti-HBs were screened from 250 volunteers by solid-phase radioimmunoassay as test subjects. Peripheral blood was centrifuged, mononuclear cells (PBMs) were extracted, 60 million PBM cells were left at 37 ° C for 1 hour, and non-adherent cells were collected and stored at low temperature. Adherent cells were treated with highly purified HBsAg (2 μg / ml) for 90 minutes at 37 ° C. After removing inhibitory T cells from non-adherent cells, they were gently poured onto antigen-treated monolayer of adherent cells, supplemented with pokeweed mitogen (PWM, 1: 200 dilution) or HBsAg / ml) and PWM (1: 200 dilution) for 5 days. Only one of four donors had a weaker response to HBsAg stimulation; the other three had strong responses.