Expression of histocompatibility antigens and characterization of mononuclear cell infiltrates in normal and neoplastic colorectal tissues of humans.

Serial frozen sections were prepared from 22 colorectal carcinomas. Additional samples were obtained from the adjacent normal bowel in 10 patients, from 6 concomitant adenomas in 5 patients, and from another 4 isolated adenomas. Mononuclear cell infiltrates were stained by the indirect immunoperoxidase technique with the use of a panel of 6 mouse monoclonal antibodies to human leukocyte antigens. The degree of infiltration was graded from 4 (heavy) to 0 (nil). The colorectal carcinomas and adjacent normal bowel showed an equal degree of leukocyte infiltration (HLe-1), graded 3-4 in 8 cases and 2-3 in the other 2 cases. In 7 carcinomas cytotoxic-suppressor T-lymphocytes (UCHT-4) graded 2-3 predominated over helper T-cells (OKT-4) graded 0-1. By contrast, in the adjacent normal bowel cytotoxic and helper cells were present in equal numbers. Among the adenomas leukocyte infiltration was grade 4 in 9 and grade 3 in 1. In 9 of the 10 adenomas cytotoxic cells graded 2 predominated over helper cells graded 0-1. The number of helper cells was equivalent among 6 concomitant adenomas and carcinomas from 5 patients. Adenomatous epithelial cells expressed class II major histocompatibility complex antigens (OKIa-1). However, carcinomatous or normal epithelium showed only faint staining with OKIa-1. The similarity in cell infiltration is consistent with an adenoma-carcinoma sequence. The predominance of cytotoxic cells in carcinomas that expressed class I major histocompatibility complex supports the association between lymphocyte infiltration and a favorable prognosis.

Proliferative and metastatic potential of exfoliated colorectal cancer cells.

Exfoliated colorectal carcinoma cells were obtained by lavage of 27 freshly resected tumor-bearing segments of human bowel with the use of either Hartmann's solution or medium 199. The tumor cells were isolated from the lavage fluid on Nycodenz (Nyegaard, Oslo, Norway) columns. Their proliferative and metastatic potentials were investigated by their ability to incorporate tritiated thymidine ([3H]dThd) and by their ability to form experimental pulmonary tumors following iv injection into immune-deprived stain A mice. Tumor cells from 7 of 12 patients incorporated [3H]dThd as detected by autoradiography of the cells. Pulmonary nodules of colorectal carcinoma were seen in a single mouse after iv injection of tumor cells from 6 of 17 patients and were histologically distinct from spontaneous lung cancers seen in 3 other animals. No tumors were seen in 12 immune-deprived mice receiving tissue culture medium 199 only. Thus exfoliated colorectal carcinoma cells can undergo further division and might give rise to implantation metastases in humans.

Inhibition of protein synthesis, pulmonary localization and pulmonary tumour formation by drug-treated tumour cells as a means of predicting their chemosensitivity.

Mouse mammary carcinoma cells were exposed in vitro to increasing concentrations of doxorubicin hydrochloride [adriamycin (ADR)] or 5-fluorouracil (5-FU). Uptake of [75Se]selenomethionine (75SeM) in a methionine-deficient medium measured the resulting inhibition of protein synthesis by the tumour cells. This was compared with the ability of the 75SeM labelled tumour cells to localize in mouse lungs and to form pulmonary tumours following intravenous (i.v.) injection into isogenic hosts. These parameters were also related to the ability of the drugs to inhibit pulmonary tumour formation in vivo when injected into mice which had received tumour cells i.v. Results from five different tumours were pooled for analysis. At the highest drug concentration (10 micrograms/ml ADR, 100 micrograms/ml 5-FU) inhibition of protein synthesis was significantly related to the in vivo action of the drugs in limiting formation of pulmonary tumors (P less than 0.02 using the rank difference coefficient). There was also a direct relationship between pulmonary localization of tumour cells following exposure to drugs, their ability to form tumour nodules (P less than 0.025) and the in vivo action of the drugs in inhibiting tumour formation (P less than 0.05). Thus inhibition of protein synthesis in vitro and pulmonary localization following i.v. injection may be of value in predicting the in vivo effect of cytotoxic drugs.

The effect of alpha and gamma interferon on cell growth and histocompatibility antigen expression by human renal carcinoma cells in vitro.

Tumour cells were separated from 19 renal carcinomas and cultured in vitro. The effect of interferon (IFN) alpha and gamma on cell proliferation was measured and compared to the effect of IFN on the expression of class I and class II major histocompatibility complex (MHC) antigens. When tested within the first 14 days of culture, IFN-alpha inhibited protein synthesis in 12 of 15 and IFN-gamma in four of nine tumours. Reduction in cell counts was in parallel. In six tumours the culture period was extended and in all six the effect of IFN-alpha was lost. Exposure to IFN-alpha induced or enhanced class I antigen expression in eight of 19 tumours and class II expression in two of 19. The analogous figures for IFN-gamma were five and three tumours. In four of five cases where a comparison could be made there was a correlation between the effects of IFN-alpha on cell proliferation and class I antigen expression. The efficacy of IFN in the treatment of renal carcinomas may thus, in part, result from inhibition of cell proliferation and enhancement of antigen expression.

Expression of major histocompatibility complex (MHC) antigens and their loss on culture in renal carcinoma.

Biopsies from the tumour and the adjacent normal kidney were obtained from 15 patients with renal cell carcinoma (RCC). The proximal convoluted tubules from which the tumour arose expressed major histocompatibility complex (MHC) class I antigen (Ag) in 3 cases and class II in none. By contrast, the carcinoma cells expressed class I Ag in 14 cases and class II Ag in 5 cases. Cells from each carcinoma were established in culture. As the culture period increased, cells from six of eight RCC showed diminished expression of class I Ag and five of six reduced expression of class II Ag. This is similar to the relative loss of class I Ag in synchronous metastases from RCC.

Verapamil enhances doxorubicin activity in cultured human renal carcinoma cells.

Cells from 22 renal cell carcinomas (RCC) were established in culture. Sensitivity of the tumour cells to doxorubicin alone and in combination with racemic verapamil, which reverses multidrug resistance, was tested using a [75Se]selenomethionine uptake assay to measure protein synthesis. The effect of verapamil was expressed as a potentiation index: LD50doxorubicin/LD50doxorubicin + verapamil. The potentiation index in 15 of these carcinomas was determined for cells within the first 14 days of culture. At 3.3 mumol/l concentration of verapamil, of the tumours sensitive to doxorubicin alone (LD50 < 0.75 microgram/ml) five of seven showed a potentiation index of > 2. For the less sensitive tumours the analogous proportion was seven of eight. Tumour cell expression of glycoprotein P-170, associated with multidrug resistance, was estimated using the monoclonal antibody C-219. Initial expression levels were unrelated to the action of verapamil. In five tumours the proportion of cells expressing P-170 declined as the period of culture increased. This was not associated with any consistent change in the LD50 for doxorubicin or in potentiation of doxorubicin sensitivity by verapamil. Cell cloning associated with prolonged cell growth in vitro could mimic tumour cell cloning which accompanies the formation of metastases. Thus reduced expression of P-170 on prolonged cell growth in vitro may be a pointer to the efficacy of combination therapy in the treatment of patients with metastatic renal cell carcinoma.

Relevance of the immune system in human urological malignancies: prospective for future clinical treatments.

The possible role of the immune system in resisting human malignancies has long been debated. Several recent findings from animal and human studies have restimulated interest in the immune surveillance hypothesis for tumor control. These findings have been complied from various disciplines including cytokine therapy, adoptive immunotherapy, and gene therapy. Following the initial euphoria, it is now clear that immunotherapy of selected cancer cases in the early stages of tumor development may make an important contribution to tumor control, particularly in dealing with minimal residual disease after tumor debulking. This review discusses some of these issues and proposes approaches that could pave the way for better selection of the patients best suited for immunotherapy. We would argue that therapies directed at the re-expression of major histocompatibility complex (MHC) class I antigens might improve outcomes in immune-therapy-based treatments.

High-dose intravenous estrogen therapy in advanced prostatic carcinoma. Use of serum prostate-specific antigen to monitor response.

High-dose intravenous estrogen therapy was shown to be effective in relieving bone pain due to metastatic disease in 22 of 29 (75.9%) men with advanced hormone-resistant prostate cancer. This clinical response was accompanied by significant falls in serum prostate-specific antigen (PSA) levels in 13 (44.8%) patients. It is suggested that this clinical benefit is due to a direct inhibitory effect of estrogen on prostate cancer cells.

Transfer of adoptive immunity by intra-arterial injection of tumor-immune pig lymph node cells: treatment of recurrent urinary bladder carcinoma after radical radiotherapy.

Twenty-four patients with invasive transitional cell carcinoma of the urinary bladder, recurrent after radiotherapy, were treated by intra-arterial infusion of tumor-immune pig lymph node cells. In 11 patients there was a remission of the disease process, and 3 patients remain alive and free of disease in excess of one year after treatment.

Treatment of bladder carcinoma with tumor-immune pig lymph node cells. Phase I study.

A total of 77 patients with invasive TCC of the urinary bladder each received a single arterial injection of tumor-immune pig lymph node cells (LNC), into the tumor blood supply. The patients were divided into 4 groups: those receiving pig LNC as the only treatment (16 patients); those who received pig LNC on relapse (judged by EUA cystoscopy and biopsy) following radical radiotherapy, 5,500 cGy (34 patients); those who received pig LNC followed after an interval of six weeks by radiotherapy, 4,000 cGy (10 patients); as in Group 3 but with the dose of radiotherapy increased to 5,500 cGy (17 patients). Complete remission was characterized by complete disappearance of the tumor (for a varying time) following treatment. Partial remission was defined as: a reduction in the level of symptoms and a decrease in tumor size on EUA and/or cystoscopy. There were 1 complete and 3 partial remissions among the patients in Group 1, 5 complete and 7 partial in Group 2, 6 complete and 2 partial in Group 3, and 8 complete and 1 partial in Group 4. Of the 20 patients showing complete remission, 7 lived for more than five years after treatment.

Comparative assessment of MHC antigen expression in bladder and testis tumour biopsies and established urological tumour cell lines: the relevance of cytokine and gene transfection for correction of defective MHC antigens.

The aim of this study using radio-binding (RB), peroxidase-anti-peroxidase (PAP) and immunoprecipitation (IP) techniques was to investigate the pattern of major histocompatibility complex (MHC) antigen expression in urological malignancies and to compare the results with those seen in established urological human tumour cell lines. The results showed that using PAP technique, the percent bladder cases showing complete loss or cases with greater than 90% of tumour cells negative with W6/32 (detects all class I antigens), HC10 (detects free heavy chain) and BMM.1 (detects beta2-mirogobulin) monoclonal antibodies (Mab) were 16%, 44% and 2% respectively. In a subgroup of 37 cases, the intensity of MHC class II antigen expression for strong, weak and negative cases were 9 (24%), 8 (22%) and 20 (54%) respectively. The expression for class I antigens on testis tumours was mainly negative and when positive, it was present in a small percent of tumour cells. This was also observed for class II antigens where only 8% of cases showed some degree of positivity. Using RB technique, 10 of 12 (85%) of tumour lines expressed class I antigens spontaneously and following interferon gamma (IFNgamma) stimulation, the 2 negative lines one testis (Tera I) and one bladder (Fen) remained negative and 2 lines (both testis lines Tera II and Ep2102) showed a significant class I up-regulation. None of the lines expressed class II antigens spontaneously and following IFNgamma stimulation, 8 of 12 (66%) were induced. The absence of class I and II antigens in the negative lines was confirmed using IP technique. In the case of one class I negative bladder cell line i.e. Fen, the biochemical analysis showed the absence of beta2-m gene product which could not be restored by IFNgamma stimulation. However, transfection of the cells with beta2-m gene resulted in the expression of fully assembled class I antigens, indicating that the loss of antigens was due to the absence of functional beta2-m gene. These results indicated the similarity between the pattern of expression of MHC antigens on tumour biopsies and established tumour cell lines. They also demonstrated that both cytokine stimulation and gene transfection could be used to correct defective class I antigens in tumour cell lines. These approaches might have important implications for pre-selection of bladder cancer patients for cytokine or gene therapies.