Osanto S, Schiphorst P P, Weijl N I, Dijkstra N, Van Wees A, Brouwenstein N, Vaessen N, Van Krieken J H, Hermans J, Cleton F J, Schrier P I
Department of Clinical Oncology, Leiden University Medical Center, The Netherlands.
Hum Gene Ther. 2000 Mar 20;11(5):739-50. doi: 10.1089/10430340050015635.
Thirty-three metastatic melanoma patients were vaccinated according to a phase I-II study with an allogeneic melanoma cell line that was genetically modified by transfection with a plasmid containing the gene encoding human interleukin 2 (IL-2). The cell line expresses the major melanoma-associated antigens and the HLA class I alleles HLA-A1, -A2, -B8, and Cw7. All patients shared one or more HLA class I alleles with this cell line vaccine. Patients were immunized by three vaccinations, each consisting of 60 x 106 irradiated (100 Gy) melanoma cells (secreting 120 ng of IL-2/10(6) cells/24 hr) administered subcutaneously at weekly intervals for 3 consecutive weeks. Side effects of treatment consisted of swelling of locoregional lymph nodes and induration at the site of injection, i.e., a delayed-type hypersensitivity (DTH) reaction. In three patients, vaccination induced inflammatory responses in distant metastases containing necrosis or apoptosis along with T cell infiltration. Apoptosis occurred only in Bcl-2-negative areas, not in Bcl-2-expressing parts of the metastases. Two other patients experienced complete or partial regression of subcutaneous metastases. Seven patients had protracted stabilization (4 to >46 months) of soft tissue metastases, including one patient who developed vitiligo after vaccination. Immune responses to the vaccine could be detected in 67% of the 27 patients measured. Vaccination was shown to induce a variable change in the number of anti-vaccine cytotoxic T lymphocytes (CTLs) in peripheral blood, which did not correlate with response to treatment. However, in two of five patients the frequency of anti-autologous tumor CTLs measured was significantly higher than before vaccination. This study demonstrates the feasibility, safety, and therapeutic potential of vaccination of humans with allogeneic, gene-modified tumor cells, and that frequencies of vaccine-specific CTLs among patient lymphocytes can be determined by using a modified limited dilution analysis (LDA).
33例转移性黑色素瘤患者按照一项I-II期研究进行疫苗接种，该研究使用的是一种经基因改造的同种异体黑色素瘤细胞系，改造方法是用含有编码人白细胞介素2（IL-2）基因的质粒进行转染。该细胞系表达主要的黑色素瘤相关抗原以及HLA I类等位基因HLA-A1、-A2、-B8和Cw7。所有患者与这种细胞系疫苗共享一个或多个HLA I类等位基因。患者接受三次疫苗接种，每次接种由60×10⁶经照射（100 Gy）的黑色素瘤细胞组成（每10⁶个细胞/24小时分泌120 ng IL-2），每周皮下注射一次，连续注射3周。治疗的副作用包括局部区域淋巴结肿大和注射部位硬结，即迟发型超敏反应（DTH）。在3例患者中，疫苗接种在含有坏死或凋亡以及T细胞浸润的远处转移灶中诱导了炎症反应。凋亡仅发生在Bcl-2阴性区域，而非转移灶中表达Bcl-2的部分。另外2例患者的皮下转移灶出现完全或部分消退。7例患者的软组织转移灶长期稳定（4至>46个月），其中1例患者在接种疫苗后出现白癜风。在接受检测的27例患者中，67%可检测到对疫苗的免疫反应。疫苗接种显示可在外周血中诱导抗疫苗细胞毒性T淋巴细胞（CTL）数量的可变变化，这与治疗反应无关。然而，在5例患者中的2例中，所检测的抗自体肿瘤CTL频率显著高于接种疫苗前。本研究证明了用同种异体、基因改造的肿瘤细胞对人类进行疫苗接种的可行性、安全性和治疗潜力，并且可以通过使用改良的有限稀释分析（LDA）来确定患者淋巴细胞中疫苗特异性CTL的频率。
