HUMAN TUMOR-SPECIFIC ANTIBODIES PRODUCED IN SCID MICE

拟议研究的总体目标是评估和完善 优化人单克隆抗体效率的新方法 （MoAb）的生产，特别是对人肿瘤具有特异性的MoAb- 相关抗原（TAA）。 我们的一般方法将是生成人类 TAA特异性免疫反应与人淋巴细胞原位， 稳定移植到C.B-17 scid小鼠中（Mosier，et al.，自然335：256， 1988年）。 提出了三种策略：（a）诱导原代人 正常成年供者移植的scid小鼠的抗体应答 淋巴细胞（B）刺激scid小鼠的二次免疫应答 移植来自荷瘤患者的淋巴细胞，和（c）激活 scid小鼠移植来自 癌症患者积极免疫肿瘤细胞。 两 本提案中包括的肿瘤类型是乳腺癌和恶性肿瘤。 胶质瘤 根据，scid小鼠将被移植来自 正常供体和乳腺癌或恶性神经胶质瘤患者，和 移植的SCID小鼠将用纯化的乳腺免疫 癌TAa糖蛋白（TAG 12或HER-2/neu）或适当的人 肿瘤细胞系（TAG 12阳性T47-D和HER-2/neu阳性SK-BR-3细胞 对于乳腺癌或用于III期活性药物的U-251 MG胶质瘤细胞系， 恶性神经胶质瘤免疫试验）。 移植和多重免疫 将监测SCID小鼠的人免疫球蛋白的血清水平， 人抗体与相应免疫原的反应性。 免疫人 将从小鼠中回收淋巴细胞并用于与B的融合 淋巴母细胞样细胞系以产生分泌人MoAb的杂交瘤。 然后分析人MoAb对人MoAb的结合特异性。 TAA和各种正常和肿瘤组织。 对这种新方法的评价 从scid小鼠体内的人淋巴细胞中产生人单克隆抗体 可以促进用于治疗的人单克隆抗体的产生增加 应用于人类恶性肿瘤，并可能产生新的信息， 抗体库在人类对肿瘤的免疫应答中的作用。

The overall objective of the proposed research is to evaluate and refine a novel approach for optimizing the efficiency of human monoclonal antibody (MoAb) production, particularly MoAb with specificity for human tumor- associated antigens (TAA). Our general approach will be to generate human TAA-specific immune responses in situ with human lymphocytes that have been stably engrafted into C.B-17 scid mice (Mosier, et al., Nature 335:256, 1988). Three strategies are proposed: (a) to induce primary human antibody responses in scid mice engrafted with normal adult donor lymphocytes, (b) to stimulate secondary immune responses in scid mice engrafted with lymphocytes from tumor-bearing patients, and (c) to activate "hyperimmune" lymphocytes in scid mice engrafted with lymphocytes from cancer patients who have been actively immunized with tumor cells. The two types of tumors included in this proposal are breast cancer and malignant glioma. According, scid mice will be engrafted with lymphocytes from normal donors and from patients with breast cancer or malignant glioma, and the engrafted scid mice will be immunized with either purified breast carcinoma TAa glycoproteins (TAG12 or HER-2/neu) or an appropriate human tumor cell line (TAG12-positive T47-D and HER-2/neu-positive SK-BR-3 cells for breast carcinoma or the U-251MG glioma line used in a phase III active immunization trial for malignant glioma). Engrafted and multiply immunized scid mice will be monitored for serum levels of human immunoglobulins and human antibody reactivity with the respective immunogen. Immunized human lymphocytes will be retrieved from mice and used in fusions with B lymphoblastoid cell lines to generate hybridomas secreting human MoAb. Human MoAb will then be analyzed for binding specificities against human TAA and various normal and tumor tissues. Evaluation of this novel method of generating human MoAb derived from human lymphocytes hosted in scid mice may facilitate increased production of human MoAb for therapeutic applications in human malignancies and may yield new information on the antibody repertoire in human immune responses to tumors.