Serum induced lymphoid cell mediated cytotoxicity to human transitional cell carcinomas of the genitourinary tract.

In some serums of patients with transitional cell carcinoma (TCC), a factor is present which induces lymphocytes from most donors with or without TCC to become cytotoxic against TCC-derived target cells. The induced cytotoxicity was directed against target cells derived from TCC's of the renal pelvis, ureter, and urinary bladder, but not against cells derived from normal kidney, bladder, testis, or skin or from renal cell carcinoma. Cytotoxicity occurred without complement but did not occur without effector cells.

Human cell-mediated cytotoxicity estimated by lymphocyte titration.

Human lymphocyte immunity to tumor-derived target cells was estimated by titrating lymphocyte concentration to achieve a 50% reduction of target cell survival. This lymphocyte titration assay gave estimates of cytotoxicity that were different from those obtained with the conventional cell-mediated cytotoxicity assays but were more proportional to lymphocyte activity. Estimates of cytotoxicity obtained using the lymphocyte titration assay were reproducible upon repeated testin over the course of several months and were relatively unaffected by two- to fourfold variations in target cell concentration. Target cell-specific cytotoxicity was reproducible but often did not appear to be tumor specific.

Classification of the bladder cancer patient based on in vitro measurements of the immune response.

In vitro measurements of the immune response in patients with cancer can be divided into those that estimate nonspecific and those that estimate tumor-specific immune responses. Contained herein is a review of these measurements, especially as they relate to studies that have been reported in patients with transitional cell carcinoma (TCC). In vitro tumor-specific immunity has been extensively examined in TCC using the lymphocyte-mediated microcytotoxicity assay, but subsequent observations on this assay have seriously jeopardized the validity of those early findings. Recent modifications of this assay have permitted longitudinal studies of lymphocyte cytotoxicity in TCC patients, and clinical correlations suggest that this modified assay may detect important immunological events. To date, however, a clinically useful classification of the TCC patient based on in vitro measurement of immune responses has not been achieved, although many promising areas still require investigation.

Placental alkaline phosphatase as a tumor marker for seminoma.

A sensitive and specific enzyme-linked immunoabsorbent assay was used in a retrospective study of serum levels of placental alkaline phosphatase (PLAP) in testicular cancer. Sixteen of 28 men with active seminoma had elevated PLAP levels, and 71% had elevated levels of either PLAP, human chorionic gonadotropin, or both. Only four of 22 men with active nonseminomatous cancer had elevated PLAP levels, and the levels were normal in all control patients, including 33 men apparently cured of testicular cancer. In six of ten serial studies, PLAP levels provided information not otherwise available that would have been useful clinically, and the levels never were elevated inappropriately. Our data suggest that PLAP is a clinically useful serum tumor marker for seminoma.

Cell line derived from a metastasis of a human testicular germ cell tumor.

A cell line, designated 833K-E, has been established from a metastasis of a human testicular germ cell tumor that consisted of four histological types of tumor cells. The 833K-E cells have morphological and ultrastructural characteristics of epithelial cells and a hyperdiploid karyotype indicative of their human male origin. The cells grow in agar cultures and produce in nude mice tumors which have the hstological features of embryonal carcinoma without differentiated elements. Many of the cells express a stage-specific mouse embryonic antigen, and low levels of the major histocompatibility antigens and beta 2-microglobulin also were detected on a large percentage of the cells. A lymphoblastoid cell line (833K-LC) established from the same tumor specimen expresses major histocompatibility antigens and beta 2-microglobulin but does not express the embryonic antigen.

Acute changes of alpha-fetoprotein and human chorionic gonadotropin during induction chemotherapy of germ cell tumors.

Thirty-seven consecutive patients with germ cell cancers (34 testicular, three extragonadal) and elevated serum levels of alpha-fetoprotein (AFP) and/or human chorionic gonadotropin (HCG) were treated with vinblastine, bleomycin, and cis-diamminedichloroplatinum induction chemotherapy. The AFP and/or HCG normalized in 36 patients. The AFP half-life was 7.9 days between Days 1 and 21 postchemotherapy but 6.0 days between Days 21 and 42 (p less than 0.05). The prolonged AFP half-life between Days 1 and 21 was related to a median increase of 57.5% (range, 22 to 219%) in the AFP level. This increase in the marker value occurred between Days 2 and 9 of therapy (median, Day 5). There was also a median increase of 181% (range, 27 to 600%) in the HCG level at a median of 5 days after the start of therapy. The increase in the AFP and HCG levels occurred in 63 and 70% of evaluable patients, respectively, and correlated with the presence of a large volume of metastatic disease (chi 2 = 8.87). Patients with relapsed or refractory disease had prolongation of the AFP half-life between Days 21 and 42 as compared to nonrelapsed patients. AFP and HCG half-life calculations should be used in the management of patients with germ cell cancers.

Localization of human renal cell carcinoma xenografts with a tumor-preferential monoclonal antibody.

We previously described an immunoglobulin G1 monoclonal antibody (UMVA-RCC-A6H) that is highly reactive with human renal cell carcinoma (RCC) and has little cross-reactivity to other cell types both normal and malignant. In efforts detailed herein, radiolabeled A6H selectively localized to RCC xenografts and provided high resolution images of the xenografts. Also, A6H clearly discriminated between RCC xenografts and other human tumor xenografts. Consistent images of RCC xenografts (greater than 60 mg) were obtained without background subtraction. The amount of radiolabeled A6H in the tumor usually ranged from five to twenty times that of the blood. Normal mouse tissues, abscesses, and other human tumor xenografts contained less radiolabel per mg than did blood. A control monoclonal antibody of the same isotype failed to exhibit any localization in xenografts or normal tissues. Approximately 40% of the radiolabeled A6H dose per g was localized in the RCC xenograft 2 days after injection, although at the time of imaging about 60% of the radiolabel remaining in the mouse was associated with the xenograft. These results demonstrate that a RCC restrictive monoclonal antibody does specifically localize to RCC xenografts and supports the hope that this approach may have clinical value for diagnosis, staging, or treatment.

Monoclonal antibodies to human renal cell carcinoma: recognition of shared and restricted tissue antigens.

Monoclonal antibodies (MABs) reactive with human renal cell carcinoma (RCC) were generated following immunization of mice with either RCC homogenates, RCC cell lines, or fetal kidney homogenates. The characteristics of four highly reactive immunoglobulin G1 MABs, designated UMVA-RCC-A6H, UMVA-RCC-A36, UMVA-RCC-C5H and UMVA-RCC-D5D are presented. The screening process consisted of a cell binding enzyme-linked immunosorbent assay and immunohistological examination of tumor, normal, and fetal tissue sections. The MABs illustrated various degrees of antigen restriction: A6H identified an antigen common to RCC, some lung and colon carcinomas, the proximal renal tubules but no other normal tissues; A36 reacted with most human tumors, the renal tubules, and many other normal tissues; C5H reacted with nearly every human cancer but of the normal tissues, only the renal glomerulus shared this antigen; D5D was very restrictive, reacting with many although not all RCC and no other cancers or normal tissues with the exception of an occasional reactivity with a Bowman's capsule. Metastatic RCC and RCC xenografts expressed these antigens. None of the MABs participated in complement-mediated cytotoxicity. In immunoprecipitation studies with L-[methyl-3H]methionine and [3H]glucosamine-HCl metabolically labeled RCC cells, C5H was shown to be associated with an antigen of Mr 115,000.

Stage II nonseminomatous testicular cancer: a 10-year experience.

Between 1970 and 1980, 82 patients with pathologic stage II nonseminomatous germ cell testicular carcinoma were treated at the University of Minnesota. Of the 30 patients treated with a retroperitoneal lymph node dissection, 22 (77%) relapsed. Of the 18 patients treated with retroperitoneal lymph node dissection and adjuvant radiotherapy, 12 (63%) relapsed. Sixteen patients received adjuvant chemotherapy before 1976, and 14 (87.5%) relapsed. After 1976, 18 patients received adjuvant chemotherapy (11 with cisplatin) and 2 (11%) have relapsed. No patient treated with cisplatin-based adjuvant chemotherapy has relapsed. The toxicity has been modest. Cisplatin-based chemotherapy is an effective and a safe adjuvant therapy for stage II nonseminomatous germ cell testicular carcinoma.

High-dose ketoconazole in advanced hormone-refractory prostate cancer: endocrinologic and clinical effects.

High-dose ketoconazole (400 mg orally three times a day) and physiologic replacement doses of glucocorticoids (hydrocortisone, 20 mg 8 AM, 10 mg 4 PM, and 8 PM) were administered to 38 patients with advanced prostatic cancer, refractory to at least initial testicular androgen deprivation. Thirty patients were completely evaluable; six were withdrawn due to possible ketoconazole-related toxicity and were considered drug failures. Two patients were unevaluable due to intercurrent therapy or inability to maintain follow-up. Ketoconazole was generally well tolerated. Mild or moderate nausea and vomiting occurred in 37% of patients, but required dose modification or discontinuation in only three patients; no hepatic damage was seen. Five of 36 patients (14%) responded to ketoconazole as determined by palpable or radiographic tumor mass reduction of 50% or greater and normalization of acid phosphatase or bone scan. Fifty percent of patients entered were stable at 90 days. Plasma androstenedione and dehydroepiandrosterone sulfate (DHEAS) were reduced markedly in almost all patients. Plasma testosterone (T) levels were low and remained unchanged, while gonadotropins were persistently elevated. Mean plasma ketoconazole content was 6.6 micrograms/mL after 28 days of therapy. While ketoconazole with hydrocortisone does suppress plasma androgens in advanced prostatic cancer patients, this infrequently causes regression of cancer that has progressed despite adequate testicular androgen ablation.

Prolonged continuous intravenous infusion interleukin-2 and lymphokine-activated killer-cell therapy for metastatic renal cell carcinoma.

PURPOSE: Two consecutive protocols of continuous intravenous (CIV) infusion interleukin-2 (IL-2) and lymphokine-activated killer (LAK) cells were carried out in patients with metastatic renal cell carcinoma (RCC) to determine the response rate and toxicity. PATIENTS AND METHODS: In both protocols, patients received induction IL-2 at 6 x 10(6) U/m2/d on days 1 to 5, and underwent leukapheresis on days 7 to 9 at the peak of rebound lymphocytosis. LAK cells were generated by a 5-day incubation with IL-2 at 1,000 U/mL, and were infused on days 12 to 14. For the first 20 patients (protocol A), maintenance IL-2 was administered at 6 x 10(6) U/m2/d on days 12 to 16. On the assumption that less IL-2 might be required to maintain rather than to induce LAK activity, and that a longer duration of maintenance IL-2 might enhance LAK survival and function in vivo, the protocol for the subsequent 22 patients (protocol B) was altered so that the maintenance phase consisted of a lower dose of IL-2 (2 x 10(6) U/m2/d) administered for a longer period of time (days 10 to 20). RESULTS: In protocol A, there were two complete responses (CRs) and three partial responses (PRs), for a total response rate of 25%. One PR was surgically converted into a CR. The durations of the CRs are 36+, 18+, and 18+ months. Hypotension and capillary leak were most severe during maintenance, which limited the median duration of maintenance IL-2 to 4 days. In protocol B, no patient experienced severe hypotension, and the median duration of maintenance IL-2 was 9 days. Two patients exhibited a CR and seven a PR, for a total response rate of 41%. Two PRs were surgically converted to CRs. The durations of CR are 14+, 9+, 6+, and 5+ months. In both protocols, the CIV induction regimen resulted in marked rebound lymphocytosis (mean, 11,097/microL) and LAK-cell yield (mean, 18.1 x 10(10)). The cumulative response rate was 14 of 42 patients, or 33% (95% confidence interval, 19% to 47%). CONCLUSION: These results demonstrate that both protocols of CIV IL-2 plus LAK cells have substantial antitumor activity, and that a longer maintenance phase of IL-2 at a lower dose is associated with significantly less toxicity without a loss of therapeutic efficacy.

Differential expression of osteonectin/SPARC during human prostate cancer progression.

The precise mechanism(s) involved in invasion and metastasis of prostate cancer (CaP) is poorly understood. Osteonectin [ON (also known as SPARC or BM-40)] is an antiadhesive protein known to be involved in cell-matrix interactions, migration, and angiogenesis. In this report, we studied the expression of ON in human prostate cell lines, primary tumors, and metastatic foci of CaP. Reverse transcription-PCR and nonradioactive in situ hybridization (ISH) techniques were used to determine ON gene expression. Immunohistochemistry was carried out using the polyclonal antibody LF37 and/or the monoclonal antibody ON-mAb. Low to moderate levels of ON mRNA and protein were observed in glandular epithelial cells of normal tissue as well as a few primary CaPs. However, high levels of ON mRNA and protein were observed in most of the CaP metastatic foci, both osseous and nonosseous. This correlated well with our findings that multiple different CaP cell lines including four CaP cell lines derived from metastases show high levels of ON gene expression. Furthermore, ISH analyses and cell-specific reverse transcription-PCR evaluation showed that both the luminal and basal cells express the ON gene. We conclude that the differential pattern of ON expression suggests that it may play an important role in the progression of CaP.

Early tumor cell dissemination in patients with clinically localized carcinoma of the prostate.

Because a significant number of patients with pathologically organ-confined carcinoma of the prostate subsequently develop recurrent disease, metastasis may occur much earlier than previously believed. We have used a reverse transcription-PCR assay for prostate-specific antigen mRNA and an immunocytochemical staining method for cytokeratins to test this hypothesis in paired peripheral blood (PB) and bone marrow (BM) specimens from 71 patients with clinically localized disease before radical prostatectomy, 14 patients with advanced-stage carcinoma of the prostate, and 30 controls (young healthy volunteers, patients without prostate disease, and patients with benign prostatic hyperplasia). Controls were negative in BM and PB. Fifty-six% of patients with organ-confined tumors (pT2) and 73% of those with extracapsular extension (pT3) were positive in the BM versus 16% of those with pT2 tumors and 27% of those with pT3 tumors in the PB. Patients with advanced-stage disease were positive in 86% of BM versus 71% of PB. The sensitivity of the immunocytochemistry assay to detect tumor cells was lower as compared with the reverse transcription-PCR assay. The results suggest that tumor cell dissemination occurs early during disease progression. Prostate cells seem to preferentially concentrate in the BM rather than the PB, which may be due to sequestration there by homing mechanisms. As the rate of detection in the BM exceeds the proportion of patients with subsequently progressing disease, we hypothesize that only a subset of these cells can survive in the BM and evolve to clinically apparent disease.

Characterization of a novel androgen-sensitive, prostate-specific antigen-producing prostatic carcinoma xenograft: LuCaP 23.

Prostatic carcinoma has proven extremely difficult to establish as cell lines or xenografts. In this article, we describe a new series of prostate cancer xenografts propagated in athymic mice, designated LuCaP 23, developed from prostate metastases harvested at autopsy shortly after death. Tumor from three separate metastatic deposits was developed into three xenograft sublines: two from lymph node metastases (LuCaP 23.1 and 23.8) and one from a liver metastasis (LuCaP 23.12). Fluorescence in situ hybridization analysis confirms the xenografts are human. Histologically, the xenografts are comprised of columnar epithelial cells arranged in a glandular pattern. Tumor doubling times range from 11 to 21 days for the three sublines. The cells secrete large amounts of prostate-specific antigen (PSA) with PSA indices of 1.27, 1.63, and 5.21 ng/ml/mm3 for the mice bearing the LuCaP 23.1, 23.8, and 23.12 sublines, respectively. Following androgen deprivation a temporary decrease in PSA secretion and a decrease in tumor size are noted in most tumors. Eventually, the tumors become androgen independent and resume growth in castrate hosts. The degree of PSA response to castration and time to PSA nadir correlate with time to progression. Thus, unlike most existing models of prostatic carcinoma, this novel xenograft exhibits many phenotypic characteristics of clinical prostatic carcinoma, including androgen sensitivity. These properties make this xenograft an excellent model for future study.

Serum osteoprotegerin levels are increased in patients with advanced prostate cancer.

PURPOSE: Osteoprotegerin (OPG) is a soluble osteoclastogenesis inhibitor that regulates bone turnover. We reported recently that OPG protein expression is significantly increased in prostate cancer (CaP) cells present in bone metastases. The aim of this study was to determine serum OPG levels in patients at different stages of CaP and correlate the results with disease status. EXPERIMENTAL DESIGN: OPG levels were examined in patients with benign prostatic hyperplasia, clinically localized CaP, early recurrence of CaP, and advanced CaP and evidence of bone metastases. Serum OPG levels were measured by sandwich ELISA assays. The serum Crosslaps (sCTX) assay was used to quantify bone resorption in the advanced CaP group. RESULTS: Serum OPG levels were increased significantly in the advanced CaP group versus all other groups. There was no significant correlation between serum OPG levels and PSA levels either in the advanced CaP group or within any of three treatment subclasses of this group: no Tx, those not treated; Tx, those treated; and R, those treated with resorption blockers. Levels of OPG were negatively correlated with sCTX levels only in the advanced CaP Tx group. sCTX levels correlated with prostate-specific antigen levels in the advanced CaP Tx and R groups but not in the no-Tx group. CONCLUSIONS: Our data show that serum OPG levels are increased with advanced CaP. We hypothesize that OPG levels are related to CaP progression and suggest that further studies of the biological effects of OPG on CaP metastases are warranted.

Preclinical models of prostate cancer.

Prostate cancer is the most common non-skin malignancy, yet the biological and molecular characteristics of the disease remain only poorly understood. One of the many reasons for this is that there are so few animal or human laboratory models of prostate cancer. Prostate cancer rarely arises spontaneously in animals, and the human cancer is unusually hard to grow in culture or as xenografts even short-term. The prostate cancer models that do exist are basically rodent models, human cell lines, human xenografts, and gene transfer and transgenic models. Many laboratories have put great effort into developing prostate cancer models, without much success. These efforts recently have been forced to be curtailed, primarily due to a lack of funding. There is good reason to believe, however, that propagation of metastatic tissue will be much more successful. At the University of Washington, the Prostate Cancer Donor Program, organized in a manner similar to existing methods for cadaver organ harvest, has been instituted to help recruit patients for these approaches. Prospects for success of model development also have been improved because of advances in techniques for the maintenance of severe combined immunodeficiency disease mice or nude mice.

Newer applications of serum prostate-specific antigen in the management of prostate cancer.

The information contained in this article indicates that PSA will have an increasing role in the management of prostate cancer. For example, it is now essential for optimal diagnosis if prostate cancer detection is the goal. Prospects are high that more information about PSA density relationships, PSA velocity phenomenologies, and possible PSA isoforms will increase diagnostic accuracy. It would also seem that PSA will improve staging accuracy not only by better manipulation of multiple preoperative parameters (eg, cancer grade, volume, PSA, etc) but possibly by the molecular detection of minute amounts of occult prostate cancer cells in bone, blood or lymph nodes, or by improved use of immune scanning. Finally, the use of these more sophisticated staging approaches together with increasingly sensitive PSA assays and possibly androgen provocative testing might allow the prospect that the potentially curative therapies can be almost immediately assessed for efficacy, thereby increasing prospects for therapeutic progress. Finally, PSA may become even more important for manipulating hormone therapies (eg, IAS therapy) or it could form a basis for new treatments such as immune or gene therapy.

Vinblastine, bleomycin and cis-diamminedichloroplatinum in the treatment of advanced testicular carcinoma. Possible importance of longer induction and shorter maintenance schedules.

Twenty-eight patients with stage III or recurrent stage I or II testicular cancer were treated with four to six cycles of vinblastine, bleomycin and cis-diamminedichloroplatinum, and then with vinblastine maintenance for one year. With a median follow-up of more than 20.5 months for all patients, complete remission has been achieved with chemotherapy and operation in 23 (82 per cent). One of these 23 patients has had a relapse, and only three are still receiving maintenance therapy. The toxicity incurred by the extra cycles of chemotherapy in patients with extensive disease was no greater than that experienced by patients receiving smaller doses of drugs. These results confirm the high response rate reported for this combination of drugs and strongly suggest that some relapses after complete remission can be prevented by longer remission induction schedules. The value of maintenance therapy is questionable and needs further study.

Prostate cancer.

Prostate carcinoma is a growing concern in our aging society. While the disease often follows a indolent course, it is the second leading cause of cancer-related deaths in males. Prostate cancer screening is promising but remains unproven and controversial. The therapy of prostate cancer has changed little over the past 10 years. The tumor remains refractory to conventional chemotherapeutic agents. True containment of this disease will require novel strategies of diagnosis, biologic assessment, and therapy.

Association of HLA and renal cell carcinoma.

HLA-A,B,C, and DR antigen frequencies were studied in a patient population with a history of renal cell carcinoma. HLA-DR5 was found in 54% of patients, which is significantly elevated over control values (20%; p less than .01). On the other hand, HLA-A,B, and C locus antigen frequencies were not abnormal compared to controls.