Patard J J, Muscatelli-Groux B, Saint F, Popov Z, Maille P, Abbou C, Chopin D
Groupe d'Etude des Tumeurs Urologiques, Centre de Recherches Chirurgicales, Créteil, France.
Br J Urol. 1996 Nov;78(5):709-14. doi: 10.1046/j.1464-410x.1996.01928.x.
To help define the optimal protocol of Bacille Calmette-Guérin (BCG) treatment for transitional cell carcinoma (TCC) of the bladder by examining cytokine production and antigen presentation to effector cells after instillations of BCG in patients with bladder TCC.
Sixty-four urine samples from 11 patients were tested for the production of interferon gamma (IFN-gamma) using a modified commercial enzyme-linked immunosorbent assay (ELISA) kit. Urine was collected before and at intervals up to 24 h after the intravesical instillation of BCG. Immunohistological studies were also carried out using a two-step alkaline phosphatase technique to explore the expression of tumour-associated antigens (TAAs) (E7, 19A211, T138), major histocompatibility complex (MHC) molecules and lymphocyte subset infiltrates (CD3, CD4, CD8) in bladder biopsies before and 3 weeks after the completion of treatment in seven patients.
IFN-gamma was only detected 4, 6 and 8 h after instillation, with a maximum concentration at 6 h (2.9-34.7 IU/mL in 10 patients). During a 6-week course of BCG, IFN-gamma was barely detectable after the first two instillations, but gradually increased from the third instillation onwards. TAA and MHC II antigens, which were absent or faintly expressed on normal urothelial cells before treatment, were expressed strongly in five patients after treatment. The local recruitment of immunocompetent cells was detected in all patients.
These results suggest that the local immune response after the intravesical instillation of BCG can be quantified using simple ELISA tests and could be useful in defining objective criteria for rationalizing treatment (dose and duration), and in determining the relation between the immune response and anti-tumour activity. There is evidence that antigen presentation is enhanced after BCG instillations, suggesting that a T cell-MHC restricted pathway might be involved in the anti-tumour response. This study supports the search for tumour-rejection antigens in bladder cancer.
通过检测膀胱移行细胞癌（TCC）患者膀胱内灌注卡介苗（BCG）后细胞因子的产生以及抗原呈递给效应细胞的情况，帮助确定膀胱TCC的最佳BCG治疗方案。
使用改良的商用酶联免疫吸附测定（ELISA）试剂盒，对11例患者的64份尿液样本进行干扰素γ（IFN-γ）产生情况的检测。在膀胱内灌注BCG之前以及灌注后长达24小时的不同时间间隔采集尿液。还采用两步碱性磷酸酶技术进行免疫组织学研究，以探究7例患者治疗前及治疗完成后3周膀胱活检组织中肿瘤相关抗原（TAAs）（E7、19A211、T138）、主要组织相容性复合体（MHC）分子以及淋巴细胞亚群浸润（CD3、CD4、CD8）的表达情况。
仅在灌注后4、6和8小时检测到IFN-γ，6小时时浓度最高（10例患者中为2.9 - 34.7 IU/mL）。在6周的BCG疗程中，前两次灌注后几乎检测不到IFN-γ，但从第三次灌注开始逐渐升高。治疗前在正常尿路上皮细胞上不存在或微弱表达的TAA和MHC II抗原，在5例患者治疗后强烈表达。所有患者均检测到免疫活性细胞的局部募集。
这些结果表明，膀胱内灌注BCG后的局部免疫反应可以通过简单的ELISA检测进行量化，这对于确定合理治疗（剂量和持续时间）的客观标准以及确定免疫反应与抗肿瘤活性之间的关系可能有用。有证据表明BCG灌注后抗原呈递增强，提示T细胞 - MHC限制途径可能参与抗肿瘤反应。本研究支持在膀胱癌中寻找肿瘤排斥抗原。
