IMMUNOGENICITY OF SYNTHETIC TF-KLH (KEYHOLE LIMPET HEMOCYANIN) AND STN-KLH CONJUGATES IN COLORECTAL-CARCINOMA PATIENTS
IMMUNOGENICITY OF SYNTHETIC TF-KLH (KEYHOLE LIMPET HEMOCYANIN) AND STN-KLH CONJUGATES IN COLORECTAL-CARCINOMA PATIENTS
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DOI：
10.1007/bf01521345
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发表时间：
1995-09-01
影响因子：
5.8
通讯作者：
LIVINGSTON, PO
中科院分区：
文献类型：
--
作者：
ADLURI, S;HELLING, F;LIVINGSTON, PO
结直肠癌的粘蛋白过表达癌症相关的二糖--结肠癌复发风险高，但手术切除后无疾病的患者，用合成TF和sTn免疫，sTn通过两个碳巴豆基连接体共价连接到匙孔limper血蓝蛋白（KLH）。4组患者分别接受无佐剂TF-KLH、TF-KLH+免疫佐剂Deter、sTn-KLH + Deter或sTn-KLH+免疫佐剂QS-21治疗，并监测血清学应答。酶联免疫吸附试验（ELISA），斑点印迹免疫染色，抑制试验，用于确定抗体反应的合成TF和sTn表位和天然抗原，包括去唾液酸血型糖蛋白表达TF抗原，绵羊颌下粘蛋白和人结肠癌细胞系LS-C表达sTn抗原。我们的研究结果表明，含有TF或sTn-KLH缀合物加上免疫佐剂Deter，特别是QS-21的疫苗诱导针对相应合成二糖表位的高IgM和IgG抗体滴度。然而，当针对表达这些二糖表位的天然抗原进行测试时，IgM抗体显示出弱至中等的反应性，而IgG抗体几乎完全不反应。在这些结果的基础上，我们继续测试合成TF和sTn表位的修饰，以鉴定那些诱导IgM和IgG抗体的修饰，这些抗体在肿瘤粘蛋白上表达时与这些抗原更具反应性。
Mucins of colorectal carcinomas overexpress the cancer-associated disaccharides Thomsen-Friedenreich antigen (TF) and sialyl-Tn antigen (sTn), making these antigens suitable for active specific immunotherapy. Patients at high risk for recurrent colon cancer, but free from disease after surgical resection, were immunized with synthetic TF and sTn covalently attached by a two-carbon crotyl linker to keyhole limper hemocyanin (KLH). Four groups of patients were treated with TF-KLH without adjuvant, TF-KLH plus the immunological adjuvant Deter, sTn-KLH plus Deter, or sTn-KLH plus the immunological adjuvant QS-21, and the serological response was monitored. Enzyme-linked immunosorbent assay (ELISA), dot-blot immunostains, and inhibition assays were used to identify antibody responses against synthetic TF and sTn epitopes and against natural antigens, including asialoglycophorin expressing TF antigen, and ovine submaxillary mucin and the human colon cancer line LS-C expressing sTn antigen. Our results demonstrate that vaccines containing TF or sTn-KLH conjugates plus immunological adjuvants Deter and especially QS-21 induced high IgM and IgG antibody titers against the respective synthetic disaccharide epitopes. However, when tested against natural antigens expressing these disaccharide epitopes, IgM antibodies showed weak to moderate reactivity, while IgG antibodies were almost totally unreactive. On the basis of these results we are continuing to test modifications of synthetic TF and sTn epitopes to identify those that induce IgM and IgG antibodies that are more reactive with these antigens as they are expressed on tumor mucins.