Trieb K, Gerth R, Windhager R, Grohs J G, Holzer G, Berger P, Kotz R
Department of Orthopedics, University of Vienna, Austria.
Immunobiology. 2000 Jan;201(3-4):368-76. doi: 10.1016/S0171-2985(00)80091-1.
Osteosarcoma is the most frequent malignant bone tumor, mainly occurring in the second and third decade of life. Diagnosis is limited to clinical symptoms, radiology and histology, but so far no diagnostic laboratory tests are available. Heat shock proteins (hsp), highly conserved proteins performing vital intracellular chaperoning functions and preventing cells from death, have been shown to be involved in tumor immunity. We analyzed 75 sera from 23 patients with high-grade osteosarcoma, 8 patients with chondrosarcoma, 10 patients with Ewing's sarcoma, 5 patients with soft tissue sarcoma, 11 patients with benign bone tumors at the time of diagnosis and from 18 healthy controls with an indirect one-site enzyme linked immunosorbent assay (ELISA) for the presence of anti-hsp60 and 70 antibodies. In these assays 10/23 osteosarcoma patients (43%) had anti-hsp60 antibodies with a mean +/- S.D. titer of 0.382 +/- 0.243 U/ml. Only one of the 18 healthy controls (1/18, 5.6%; titer 0.22 U/ml), two of the Ewing's sarcoma patients (2/10, 20%; titer 0.2 +/- 0.09 U/ml), two of the patients with a benign bone tumor (2/11, 18%; titer 0.22 +/- 0.16 U/ml) and one of the chondrosarcoma patients (1/8, 12.5%; titer 0.14 U/ml) were positive, whereas all others, including all soft tissue sarcomas were negative throughout. Anti-hsp60 antibodies in patients with osteosarcoma are therefore significantly increased (p < 0.05). 19/23 (83%) of osteosarcoma biopsy specimens expressed hsp60 immunohistochemically and all specimens from patients with a positive anti-hsp60 serum titer expressed hsp60. The level of the anti-hsp60 antibodies did not correlate with clinical parameters such as response to preoperative chemotherapy, duration of symptoms, age, gender, tumor size, serum alkaline-phosphatase levels and metastases. Although no difference in anti-hsp70 antibodies could be observed between sera from patients and healthy controls, a positive correlation was found for the presence of anti-hsp70 serum antibodies and lung metastases at the time of diagnosis in osteosarcoma patients. These data suggest an increase of anti-hsp60 antibodies at the time of first diagnosis of osteosarcoma. These findings should therefore give rise to further investigations on a group of new markers for the diagnosis of osteosarcoma.
骨肉瘤是最常见的恶性骨肿瘤，主要发生在生命的第二个和第三个十年。诊断仅限于临床症状、放射学和组织学，但到目前为止尚无诊断性实验室检测方法。热休克蛋白（hsp）是高度保守的蛋白质，执行重要的细胞内伴侣功能并防止细胞死亡，已被证明与肿瘤免疫有关。我们用间接单位点酶联免疫吸附测定（ELISA）分析了23例高级别骨肉瘤患者、8例软骨肉瘤患者、10例尤因肉瘤患者、5例软组织肉瘤患者、11例诊断时的良性骨肿瘤患者以及18名健康对照者的75份血清中抗hsp60和70抗体的存在情况。在这些检测中，10/23例骨肉瘤患者（43%）有抗hsp60抗体，平均±标准差滴度为0.382±0.243 U/ml。18名健康对照者中只有1人（1/18，5.6%；滴度0.22 U/ml）、2例尤因肉瘤患者（2/10，20%；滴度0.2±0.09 U/ml）、2例良性骨肿瘤患者（2/11，18%；滴度0.22±0.16 U/ml）和1例软骨肉瘤患者（1/8，12.5%；滴度0.14 U/ml）呈阳性，而包括所有软组织肉瘤在内的所有其他患者均为阴性。因此，骨肉瘤患者的抗hsp60抗体显著增加（p<0.05）。23例骨肉瘤活检标本中有19例（83%）免疫组化表达hsp60，抗hsp60血清滴度阳性患者的所有标本均表达hsp60。抗hsp60抗体水平与临床参数如术前化疗反应、症状持续时间、年龄、性别、肿瘤大小、血清碱性磷酸酶水平和转移情况无关。虽然患者血清和健康对照者血清中的抗hsp70抗体无差异，但在骨肉瘤患者诊断时，抗hsp70血清抗体的存在与肺转移呈正相关。这些数据表明在骨肉瘤首次诊断时抗hsp60抗体增加。因此，这些发现应促使对一组骨肉瘤诊断新标志物进行进一步研究。
