Riddell S R, Greenberg P D
Fred Hutchinson Cancer Research Center, Seattle, WA 98104.
J Immunol Methods. 1990 Apr 17;128(2):189-201. doi: 10.1016/0022-1759(90)90210-m.
Antigen-specific T cell clones are useful reagents for studies of the fine specificity of antigen recognition and of potential therapeutic use in adoptive immunotherapy for human viral and malignant diseases. Culture methods which require antigen and APC for stimulation can be problematic for the generation and long-term growth of human virus and tumor-specific T cells. We have developed an alternative culture method using monoclonal antibodies to T cell activation molecules, CD3 and CD28, as stimulation to efficiently grow CD4+ and CD8+ antigen-specific T cells from single progenitors and expand T cell clones in long-term culture. This method alleviates the requirement for large amounts of viral or tumor antigens and MHC compatible APC to sustain the growth of virus and tumor-specific T cell clones, and, as demonstrated for CD8+ CMV-specific cytotoxic T cells, overcomes the difficulties cloning CD8+ T cells using virally infected cells as antigen-presenting cells. T cell clones generated and maintained with monoclonal antibody stimulation are rapidly expanded and retain antigen-specific responses after 3 months in culture, suggesting this approach may prove useful for growing large numbers of antigen-specific T cell clones for cellular immunotherapy.
抗原特异性T细胞克隆是研究抗原识别精细特异性以及用于人类病毒和恶性疾病过继免疫治疗潜在治疗用途的有用试剂。需要抗原和抗原呈递细胞（APC）进行刺激的培养方法对于人类病毒和肿瘤特异性T细胞的产生和长期生长可能存在问题。我们开发了一种替代培养方法，使用针对T细胞活化分子CD3和CD28的单克隆抗体作为刺激，以有效地从单个祖细胞中培养CD4+和CD8+抗原特异性T细胞，并在长期培养中扩增T细胞克隆。该方法减轻了维持病毒和肿瘤特异性T细胞克隆生长对大量病毒或肿瘤抗原以及MHC相容的APC的需求，并且如针对CD8+巨细胞病毒特异性细胞毒性T细胞所证明的，克服了使用病毒感染细胞作为抗原呈递细胞克隆CD8+ T细胞的困难。通过单克隆抗体刺激产生和维持的T细胞克隆迅速扩增，并在培养3个月后保留抗原特异性反应，表明这种方法可能被证明对为细胞免疫治疗培养大量抗原特异性T细胞克隆有用。
