HUMAN MONOCLONAL ANTIBODIES DERIVED FROM SCID MICE

这项建议的长远目标是发展一个系统， 可靠地生产高亲和力的人单克隆抗体， 理想的同种型。 第一阶段提案的具体目标是共同优化 两个实验系统：1）最佳体外条件 刺激人脾淋巴细胞和2）的特异性， 转移和再刺激这些人淋巴细胞在严重的组合 免疫缺陷（SCID）小鼠，因此高亲和力IgG同种型人 可以更容易地获得单克隆抗体。 具体来说，我们将 1)优化人脾细胞的体外刺激以转移至 SCID小鼠; 2）优化体内抗原再刺激SCID小鼠的条件; 这些人脾细胞在SCID小鼠中;和3）确定是否额外的 共施用给SCID小鼠的人淋巴组织或使用双 突变的SCID + beige小鼠可以提高移植物的抗体应答。 人脾细胞 高亲和力人单克隆抗体的商业应用 包括被动免疫、肿瘤成像、癌症免疫治疗， 免疫缺陷和过敏的治疗。 人单克隆抗体 将减轻对传染性污染物的担忧， 当从捐献者的血液中制备出人类抗体时， 他们将 允许产生大量明确定义的抗体。 他们 在病人体内的半衰期应该比小鼠抗体长。 期望 效应器机制应该得到最佳刺激，因为人类 来源和单克隆抗体提供的同种型控制。 最后，它们将最大限度地减少在管理 抗人的外源抗体，特别是人抗小鼠抗体（HAMA） 这一反应最近引起了广泛关注。

The long term objective of this proposal is to develop a system for the reliable production of human monoclonal antibodies of high affinity and desirable isotype. The specific aims of this phase one proposal are intended to co-optimize two experimental systems: 1) The conditions for optimal in vitro stimulation of human splenic lymphocytes and 2) The specifics of the transfer to and restimulation of these human lymphocytes in severe combined immunodeficient (SCID) mice, so that high affinity IgG isotype human monoclonal antibodies can be more readily obtained. Specifically, we will 1) optimize the in vitro stimulation of human spleen cells for transfer to SCID mice; 2) optimize the conditions for in vivo antigen re-stimulation of these human spleen cells in SCID mice; and 3) determine whether additional human lymphoid tissue co-administered to the SCID mice or the use of double mutant SCID + beige mice can improve the antibody response of the grafted human spleen cells. Commercial applications of high affinity human monoclonal antibodies include passive immunization, tumor imaging, cancer immunotherapy and treatment of immunodeficiencies and allergies. Human monoclonal antibodies will alleviate much of the concern over infectious contaminants which arises when human antibodies are prepared from donor blood. They will allow the production of large quantities of well-defined antibodies. They should have longer half lives in patients that mouse antibodies. Desired effector mechanisms should be optimally stimulated because of the human origin and the control of isotype which monoclonal antibodies provide. Finally, they will minimize the problems encountered when administering a foreign antibody to humans, especially the human anti-mouse antibody (HAMA) response which has received much recent attention.