Jurincic-Winkler C, Metz K A, Beuth J, Engelmann U, Klippel K F
Department of Urology, General Hospital Celle, Germany.
Br J Urol. 1995 Dec;76(6):702-7. doi: 10.1111/j.1464-410x.1995.tb00760.x.
To determine whether keyhole limpet haemocyanin (KLH) instilled intravesically improves the local cellular response within the bladder wall of patients suffering from superficial bladder carcinoma.
Twelve patients (10 men and two women, mean age 67 years, range 42-85) with superficial carcinomas of the bladder were treated for 6 consecutive weeks and then monthly for 1 year with 20 mg KLH in 20 mL saline instilled intravesically after complete resection of the tumours. Biopsies were taken for immunohistochemical examination before treatment and again 6 weeks, 3, 6 and 9 months after treatment. Six patients with no evidence of cystitis or malignant bladder disease acted as a control group. Immunofluorescent staining of the biopsies was performed using monoclonal antibodies to the T-cell markers CD4 and CD8, and to CD14 (monocytes), CDw15 (granulocytes), CD19 B-cells (Pan-B), CD68 (macrophages) and HLA-DR. Anti-KLH antibody-producing plasma cells were detected using a standard technique. A semiquantitative analysis of locally infiltrating cell types was performed.
After treatment with KLH the increase of CD8+ suppressor cells was less pronounced than that of CD4+ helper cells. The T-helper/inducer to T-suppressor/cytotoxic cell ratio thus altered from 0.8:2.0 before treatment to 1.6:2.3 afterwards. Hence, the number of T-helper cells had increased considerably, whereas there was only a moderate increase in the number of T-suppressor cells. This cellular ratio could be detected for 9 months after KLH therapy. The numbers of activated HLA-DR+ immune cells in the submucosa and among urothelial cells also increased after KLH instillation. The degree of mononuclear cell infiltration of the submucosa increased considerably, but granulocyte infiltration was only moderate. Lymph follicles with enhanced B-lymphocyte counts were also detected.
Immune-cell infiltration into the urothelium and enhanced activation (expression of class II antigens) suggests distinct processes of cellular antigen recognition, which could be detected for up to 9 months after the beginning of KLH therapy. This may represent a basic functional mechanism of KLH therapy.
确定膀胱内灌注钥孔戚血蓝蛋白（KLH）是否能改善浅表性膀胱癌患者膀胱壁内的局部细胞反应。
12例（10例男性，2例女性，平均年龄67岁，范围42 - 85岁）浅表性膀胱癌患者在肿瘤完全切除后，用20mg KLH溶于20mL生理盐水中膀胱内灌注，连续治疗6周，之后每月1次，共治疗1年。在治疗前以及治疗后6周、3个月、6个月和9个月进行活检，用于免疫组化检查。6例无膀胱炎或膀胱恶性疾病证据的患者作为对照组。使用针对T细胞标志物CD4和CD8、CD14（单核细胞）、CDw15（粒细胞）、CD19 B细胞（泛B细胞）、CD68（巨噬细胞）和HLA - DR的单克隆抗体对活检组织进行免疫荧光染色。采用标准技术检测产生抗KLH抗体的浆细胞。对局部浸润细胞类型进行半定量分析。
用KLH治疗后，CD8⁺抑制细胞的增加不如CD4⁺辅助细胞明显。因此，T辅助/诱导细胞与T抑制/细胞毒性细胞的比例从治疗前的0.8:2.0变为治疗后的1.6:2.3。由此可见，T辅助细胞数量显著增加，而T抑制细胞数量仅适度增加。这种细胞比例在KLH治疗后9个月仍可检测到。KLH灌注后，黏膜下层和尿路上皮细胞中活化的HLA - DR⁺免疫细胞数量也增加。黏膜下层单核细胞浸润程度显著增加，但粒细胞浸润仅为中度。还检测到B淋巴细胞计数增加的淋巴滤泡。
免疫细胞浸润到尿路上皮并增强激活（II类抗原表达）提示了细胞抗原识别过程明显，在KLH治疗开始后长达9个月均可检测到。这可能代表了KLH治疗的一种基本功能机制。
