Consistent antibody response against ganglioside GD2 induced in patients with melanoma by a GD2 lactone-keyhole limpet hemocyanin conjugate vaccine plus immunological adjuvant QS-21.
Consistent antibody response against ganglioside GD2 induced in patients with melanoma by a GD2 lactone-keyhole limpet hemocyanin conjugate vaccine plus immunological adjuvant QS-21.
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发表时间：
2003-11
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通讯作者：
G. Ragupathi;P. Livingston;Chandra Hood;J. Gathuru;S. Krown;P. Chapman;J. Wolchok;L. Williams;Roberta C Oldfield;W. Hwu
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作者：
G. Ragupathi;P. Livingston;Chandra Hood;J. Gathuru;S. Krown;P. Chapman;J. Wolchok;L. Williams;Roberta C Oldfield;W. Hwu
目的：黑色素瘤、肉瘤和神经母细胞瘤在细胞表面大量表达神经节苷脂GD2，它容易受到抗体的免疫攻击。GD2在这些肿瘤上的过表达是惊人的，用针对GD2的单克隆抗体治疗神经母细胞瘤后临床反应的频率也是惊人的。此外，临床前模型已经证明gd2 -锁眼帽贝血青素（KLH）结合疫苗能够诱导消除微转移的抗体。然而，以前接种GD2-KLH患者未能诱导一致的相关抗体反应。我们在此测试使用GD2内酯- klh是否可以克服GD2- klh的低免疫原性。实验设计18例黑色素瘤患者在第0、1、2、3、10和24周接种了s.c。每组6名患者在含有GD2L- klh和免疫佐剂QS-21的疫苗中注射三种剂量水平（3、10或30微克）的GD2L内酯（GD2L）。定期抽血以评估抗体反应。结果疫苗耐受性良好。所有三种剂量水平的大多数患者产生抗gd2抗体，ELISA检测可检测到。免疫薄层色谱法也证实了GD2的特异性。虽然三组之间的滴度没有统计学差异，但30-微g剂量水平的患者通过ELISA总体上具有更高的滴度和更持久的抗体反应（IgM/IgG峰值滴度中位数为1:640/1:80），荧光激活细胞分选产生最强的细胞表面反应性（接种前和接种后血清IgM峰值阳性细胞百分比中位数为10%/63和70%/135）。接种30微g GD2剂量的患者也具有最有效的补体依赖性细胞毒性，6例患者中有5例通过荧光活化细胞分选显示出很强的细胞表面反应性，血清稀释度为1/100的铬释放显示出30%的细胞毒性。结论GD2L-KLH结合疫苗加佐剂QS-21在大多数黑色素瘤患者中诱导针对GD2的抗体结合到细胞表面并诱导补体依赖性细胞毒性。
PURPOSE Melanomas, sarcomas, and neuroblastomas abundantly express the ganglioside GD2 on the cell surface where it is susceptible to immune attack by antibodies. Overexpression of GD2 on these tumors is striking, as is the frequency of clinical responses after treatment of neuroblastoma with monoclonal antibodies against GD2. In addition, preclinical models have demonstrated the ability of a GD2-keyhole limpet hemocyanin (KLH) conjugate vaccine to induce antibodies that eliminate micrometastases. However, vaccination of patients with GD2-KLH has previously failed to induce a consistent relevant antibody response. We test here whether the use of GD2 lactone-KLH can overcome the low immunogenicity of GD2-KLH. EXPERIMENTAL DESIGN Eighteen patients with melanoma were vaccinated s.c. in the adjuvant setting on weeks 0, 1, 2, 3, 10, and 24. Groups of 6 patients were entered at three dose levels (3, 10, or 30 micro g) of GD2 lactone (GD2L) in vaccines containing GD2L-KLH plus the immunological adjuvant QS-21. Blood was drawn at regular intervals to assess the antibody response. RESULTS The vaccine was well tolerated. The majority of patients in all three dose levels produced anti-GD2 antibodies detectable by ELISA assay. Specificity for GD2 was also confirmed by immune thin-layer chromatography. Although there was no statistical difference in terms of titers between the three groups, patients at the 30- micro g dose level had higher titers and longer lasting antibody responses overall by ELISA (median IgM/IgG peak titer 1:640/1:80) and generated the strongest cell surface reactivity by fluorescence-activated cell sorting (median IgM peak percentage positive cells/mean fluorescence intensity for pre- and postvaccination sera is 10%/63 and 70%/135). Patients vaccinated with the 30- micro g GD2 dose also had the most potent complement dependent cytotoxicity using human complement, with 5 of 6 patients showing strong cell surface reactivity by fluorescence-activated cell sorting and >30% cytotoxicity by chromium release with a serum dilution of 1/100. CONCLUSIONS GD2L-KLH conjugate vaccine plus adjuvant QS-21 induces antibodies against GD2 that bind to the cell surface and induce complement-dependent cytotoxicity in the majority of patients with melanoma.