INTERACTION OF GZ-SPECIFIC T CELL CLONES IN SUPPRESSION

本申请中概述的研究建议调查以下部分 β-半乳糖苷酶(GZ)特异性T-T细胞体外抑制作用 互动。为此，T辅助细胞(Th)和T抑制细胞(Ts) 将建立克隆。由于关键的抗原决定簇，与 不同的帮助者和抑制者功能，已经被证明存在于 四聚体(1021个氨基酸残基/单体)大肠杆菌GZ，溴化氰 含有这些决定因素的多肽将用于体内诱导 CBA/J小鼠的Th、Ts和T抑制诱导因子(TSI)细胞。 随后，这些细胞将被培养以获得T细胞克隆 具有各自的功能和抗原性。已建立的T细胞 然后将克隆用于刺激的GZ特异性抑制培养 荧光素(FITC)衍生的GZ(GZ-FITC)用于抗FITC 斑块形成细胞(PFC)反应。事实上，每个功能T细胞 SET属于在GZ分子上表达的唯一表位特异性 允许仔细检查T细胞通讯的性质。《The T》 细胞克隆方法将在研究 最低限度的细胞需求，除了B细胞，在抑制 体外抗体反应。能正确诱导Th和TS细胞克隆 从事B细胞失活所反映的抑制行为 功能，还是需要其他T细胞亚群的帮助？在……里面 特别是，对抑制剂中TSI的催化活性的要求 功能将被调查。此外，对抑制者的剖析 培养(Th-Ts-B)到不同的T-T相互作用，即Th-Ts，Th-Tsi， 将进行TS-TSI，以确定这种互动是否受到限制 通过表位特异性，因此只有某些T细胞参与作为 镇压的目标。抗原本身的调节作用可能 导致这种受限的互动过程，因为一些表位在 无论是碎裂的还是完整的抗原，都不能被相互作用的人接触到 细胞。因此，细胞和抗原之间的相遇将 也可以在T细胞克隆水平上进行分析。特别是，我们将 研究抗原是否可以直接影响T细胞而不是 由抗原提呈细胞(APC)处理和呈递。T细胞 克隆为研究目前仍知之甚少的机制提供了可能性。 免疫调节，从而有助于延续应用 临床情况下的免疫学。

The study outlined in this application proposes to investigate a segment of in vitro suppression involving Beta-galactosidase (GZ) specific T-T cell interactions. For this purpose T helper (Th) and T suppressor (Ts) cell clones will be established. Since key antigenic determinants, with distinct helper and suppressor functions, have been shown to exist on the tetrameric (1021 amino acid residues/monomer) E. coli GZ, cyanogen bromide peptides containing those determinants will be used for in vivo induction of Th, Ts, and T suppressor inducer (Tsi) cells in CBA/J mice. Subsequently, these cells will be cultured to obtain T cell clones with respective functional and antigenic specificities. Established T cell clones will then be utilized in GZ-specific suppressor culture stimulated with fluorescein (FITC) derivatized GZ(GZ-FITC) for anti-FITC plaque-forming cell (PFC) response. The fact that each functional T cell set belongs to a unique epitope specificity expressed on the GZ molecule permits careful scrutiny of the nature of T cell communications. The T cell clone approach will be particularly valuable in investigating the minimal cellular requirements, aside from B cell, in the suppression of antibody response in vitro. Can properly induced Th and Ts cell clones engage in the act of suppression reflected in an inactivation of B cell function, or do they require assistance from other T cell subsets? In particular, the requirement for a catalytic activity of Tsi in suppressor function will be investigated. Furthermore, dissection of the suppressor culture (Th-Ts-B) into distinct T-T interactions, i.e., Th-Ts, Th-Tsi, Ts-Tsi, will be done to ascertain whether such interactions are restricted by epitope specificity, and thus ohly certain T cells participate as targets for suppression. The regulatory role of the antigen itself may cause this circumscribed interactive process, in that some epitopes on either fragmented or intact antigen are not accessible to the interacting cells. Therefore, the encounter between the cells and the antigen will also be analyzed at the T cell clonal level. In particular, we will investigate whether antigen can affect the T cells directly without being processed and presented by an antigen presenting cells (APC). T cell clones offer a potential to study still poorly understood mechanism of immune regulation, and thus contribute for a continuation of applied immunology in clinical situations.