Adoptive immunotherapy with LAK cells conjugated with epidermal growth factor receptor antibody

LAK 细胞与表皮生长因子受体抗体偶联的过继免疫疗法

• 批准号: 06557111
• 负责人: TAKADA Kazuaki
• 金额: $ 3.58万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06557111/
• 关键词: Oral cancer; LAK cells; Adoptive immunotherapy; epidermal growth factor receptor; Serum-free culture; Interleakin-2; インターワイナン-2; 免疫療法; 上皮成長因子; 抗体結合LAK

We have developed a serum-free medium designated RDSF for the generation of LAK cells based on RD6F medium, which was originally developed as a serum-free medium for the growth of myeloma and hybridoma cells. The cytotoxic activity of LAK cells generated in RDSF against Raji, K562 and oral cancer cells, is 3-4 times that of LAK cells generated in medium containing 10% human type-AB serum. RDSF medium consisted of nutrient mixture supplemented with transferrin, 2-aminoethanol, 2-mercaptoethanol, sodium selenite and interleukin-2. In this study, we have found that insulin, which has been found to be the most important polypeptide hormone in surum-free media for animal cells, inhibited the generation of cytotoxic activity of LAK cells cultured from pheripheral blood lymphocytes. In addition, we have found that transferrin was an essential component for the growth and generation of LAK cells in serum-free culture. Thses results suggest that RDSF will be usefel for adoptive immunotherapy of cancer and studying factors involved in the growth and differentiatiation of LAK cells.In addition, we have studied effect of monoclonal antibody (MoAb) to EGF-r designated as 12-93 on the growth of squamous cell carcinoma cells (SCC) and salivary gland adenocarcinoma cells (SAC) in vivo, and targeting immunotherapy using 12-93 MoAb-conjugated LAK cells against oral cancer cells in vitro, A 12-93 MoAb inhibited the growth og SCC and SAC in vivo. The 12-93 conjugated LAK cells showed significantly enhanced cytolysis of 12-93 reactive A431 and HSG cells but not Raji cells which dose not react with 12-93. These results indicated that 12-93 will be aviable altemative to tumor specific MoAb and 12-93-conjugated LAK cells may provide an effective strategy for targeting adoptive immunotherapy of cancer.

我们在RD 6 F培养基的基础上开发了一种无血清培养基，命名为RDSF，用于产生LAK细胞，RD 6 F培养基最初是作为骨髓瘤和杂交瘤细胞生长的无血清培养基开发的。在RDSF中产生的LAK细胞对Raji、K562和口腔癌细胞的细胞毒活性是在含10%人AB型血清的培养基中产生的LAK细胞的3-4倍。RDSF培养基由补充有转铁蛋白、2-氨基乙醇、2-巯基乙醇、亚硒酸钠和白细胞介素-2的营养混合物组成。在本研究中，我们发现，胰岛素，这已被发现是最重要的多肽激素在无苏培养基中的动物细胞，抑制LAK细胞的细胞毒活性的产生从外周血淋巴细胞培养。此外，我们还发现转铁蛋白是LAK细胞在无血清培养中生长和增殖的重要成分。本实验还研究了抗EGF-r单克隆抗体（MoAb）12-93对鳞癌细胞（SCC）和涎腺腺癌细胞（SAC）生长的影响。12-93单克隆抗体结合LAK细胞对口腔癌细胞的体外靶向免疫治疗，12-93单克隆抗体对口腔癌细胞的体内生长有抑制作用。12-93结合的LAK细胞对12-93反应的A431和HSG细胞的杀伤作用明显增强，但对不与12-93反应的Raji细胞的杀伤作用不明显。这些结果表明，12-93是一种有效的肿瘤特异性单克隆抗体的替代物，12-93偶联的LAK细胞可能为肿瘤的过继免疫治疗提供一种有效的靶向策略。

项目成果

谷 亮治 他: "LAK療法が奏功した肺転移を併った無色素性悪性黒色腫の1例" 日本口腔外科学会誌. 40. 825-827 (1994)

Ryoji Tani 等：“用 LAK 疗法成功治疗无色素性恶性黑色素瘤伴肺转移的病例”日本口腔颌面外科杂志 40. 825-827 (1994)。

T. Okamoto et al: "Adoptive immunotierapy of oral cancer with lynplohire-activated killer(LAK)cells induced in newly developed serum-free medium." Dentistry in Japan. 32. 67-70 (1995)

T. Okamoto 等人：“在新开发的无血清培养基中诱导 Lynplhire 激活杀伤 (LAK) 细胞对口腔癌进行过继免疫治疗。”

T. OKAMOTO et al.: "Adoptive immunotherapy of Oral cancer with lyuphohine actinated Killer cells induced in neuly developed seuen-fill medin." Detistry in Japan. 32. 67-70 (1995)

T. OKAMOTO 等人：“用在新开发的 seuen-fill medin 中诱导的 lyuphohin 激活的杀伤细胞对口腔癌进行过继免疫治疗。”

谷 亮治 他: "Lymphokine-activated Killer (LAK)細胞を用いた養子免疫療法に関する研究" 日本口腔外科学会誌. 40. 852-863 (1994)

Ryoji Tani 等人：“使用淋巴因子激活杀伤细胞 (LAK) 的过继性免疫疗法的研究”日本口腔颌面外科学会杂志 40. 852-863 (1994)。

Tani, R., Okamoto, T., Sakamoto, A.: "Yabumoto, M., Toratani, S.and Takada, K.A case of an amelanotic malignant melanoma successfully treated with LAK therapy" Jpn.J.Oral Maxillofac.Surg.Vol.40, No.7. 825-827 (1994)

Tani, R.、Okamoto, T.、Sakamoto, A.：“Yabumoto, M.、Toratani, S.和 Takada, K.A 例采用 LAK 疗法成功治疗无色素性恶性黑色素瘤”Jpn.J.Oral Maxillofac.Surg。