Development of chemoimmunotherapy (CI) for AIDS patients using thiophenyl urea (TUR) & nonvirion antigen vaccine (NVA).

AIDS: This article examines the development of a chemoimmunotherapy regimen for AIDS patients in which thiophenyl urea (TUR) and nonvirion antigen vaccine (NVA) were used. The article describes each step of the process from the single testing of TUR and NVA to its combination. Results from the combination therapy show that at six months, one of two patients using the combination showed a vast improvement, and the other patient did not improve until about nine months. Lung, liver, and nodal lesions cleared gradually; HIV DNA fell to low levels, possibly a dormant state; and CD4/CD8 ratios normalized. An explanation of these results is provided.^ieng

Tumour dormancy: initiation, maintenance and termination in animals and humans.

The authors review the natural occurrence of tumour dormancy in man and animal models, which show the importance of cellular defence mechanisms in inducing and maintaining the state of tumour dormancy. Six cases of non-small-cell lung cancer are described. The patients exhibited moderate to very strong delayed hypersensitivity reactions to soluble lung-cancer antigen after specific immunotherapy. Three patients had nonregional metastases at 11, 12 and 14 years. Two had regional recurrence after 9 years, and in one patient a small hilar-node metastasis was found at autopsy after 7.6 years. In each case an immunodepressive event or drug treatment preceded resurgent growth. The effect of renal transplantation in patients with a history of surgery for cancer supports the conclusion that cellular defence mechanisms are crucial for the maintenance of tumour dormancy in man.

Adjuvant, specific, active immunotherapy for resectable squamous cell lung carcinoma: a 5-year survival analysis.

In 1976 Stewart et al. (Annals of the New York Academy of Sciences 277:436-466) reported the effectiveness of adjuvant specific active immunotherapy of lung carcinoma in improving the postoperative survival of stage I lung carcinoma patients in a phase II study using lung carcinoma-associated antigen (TAA) and complete Freund's adjuvant (CFA). A phase III study was then designed by the authors to see the effects of specific active immunotherapy compared to the conventional management (no treatment) and to nonspecific immunotherapy. From 1976 to 1981, 85 patients with resectable (stages I and II) squamous cell lung carcinoma were entered into a randomized study: 1) control group; 2) specific immunotherapy group--three monthly doses of 500 micrograms of TAA emulsified with CFA; 3) nonspecific immunotherapy group--three monthly doses of CFA emulsified in saline. All the patients in the study received skin tests with 100 micrograms of the same TAA used for the immunotherapy. Recently, a 5-year follow-up of all the patients became available: The life table 5-year survival of group 1 was 34%, of group 2 was 75%, and of group 3 was 53%. The median survivals for the three groups were group 1, 38 months; group 2, 106 months; and group 3, 71 months. The difference was significant at P = .007 (Cox-Mantel test).

Endogenous production of infectious Inoue-Melnick virus in a human meningioma cell line.

We investigated continuous production of Inoue-Melnick virus (IMV) in the MG-1 cell line, established from human meningioma. The infectious virus, identified as a type 1 virus, was mostly recovered extracellularly. Assay of MG-1 cells as infective centers indicated that most of the cells were capable of producing infectious virus. By immunofluorescence, more than 90% of the cells were found to have IMV-associated cytoplasmic antigen(s) (IMCA).

Isolation of Inoue-Melnick virus from human meningioma-derived cell cultures and detection of antibody in patients with meningioma.

Inoue-Melnick virus (IMV) was isolated from six of seven human meningioma-derived cell cultures, while the virus was not isolated from six other brain tumor cell cultures. Sera of 145 consecutive neurosurgical inpatients were tested for IMV-neutralizing antibody. Of 26 patients with meningioma, 22 were positive for IMV antibody (84.6%). Of the remaining 119 patients, 16 were positive.

Biological markers of breast cancer: a review.

This review of biological markers of breast carcinoma has summarized a fairly representative series of studies pertaining to endocrine, carcinoembryonic, tissue-associated, serologic, biochemical, and specific tumor markers, with attention to some of the contributions from genetic engineering techniques applied to this research area. It is meant to be a helpful review with two purposes: those involved in research or clinical studies of breast cancer may wish to use this review as a basis for adding additional references in each category and to refer to the references provided for further study, thought, and synthesis of the data, perhaps to stimulate new associations between pertinent facts; those involved in the general scientific or clinical fields may wish to use it as an overview of biological markers pertaining to a particular form of cancer and perhaps this review will permit faster, more easily accessible synthesis of data, and a better understanding of a variety of parameters which are associated with breast cancer diseases.

Antigenic epitopes of three tumor-associated antigens of well-differentiated squamous cell carcinoma of the larynx for use in monitoring patients in specific active immunochemotherapy trials.

Laryngeal tumor-associated antigen (LX-TAA-1) monoclonal antibody-derived epitopes were tested by enzyme immunoassay (ELISA) against a coded series of 30 non-laryngeal squamous cancer sera, 32 laryngeal squamous cancer sera and 36 non-squamous cancer sera. Epitope LX-TAA-1 a62 cross-reacted with 46% of non-laryngeal squamous and with 16% non-squamous cancer sera. Epitope i5 cross-reacted with 50% non-laryngeal squamous cancer sera and with 11% non-squamous cancer sera. LX-TAA-1 epitope f18 cross-reacted with 26% of non-laryngeal squamous cancer sera but with only one, at weaker titer, of the non-squamous cancer serums (2%). Preliminary screening had ruled out any reactivity to 92 normal, age- and sex-matched sera. One hundred percent of laryngeal squamous cancer sera showed reactivity to one or more of the three epitopes. These results from highly sensitive testing imply that epitope f18 protein is probably a unique gene product of certain squamous cancer cells in the body. Serial titrations of immunized patient sera indicated that epitope f18 produces a strong but very early reactivity and appears to be a marker for early post-immunization response in patients on immunotherapy, with epitope i5 showing a low but continuing positive response starting at 7 months post-immunization. This is in contrast to our studies in other cancer systems where earlier responses are usual.

Screening of hybridoma supernatants raised against membrane fractions from breast cancer using an immunodot assay.

Membranes were prepared from mammary tumors and fractionized using gel filtration and gradient polyacrylamide gel electrophoresis. To obtain monoclonal antibodies against tumor-associated antigens, mice were immunized against membrane fraction 2a. After hybridization, we obtained 746 hybridoma cell lines. We performed the immunodot assay for screening. Every supernatant was tested against 15 antigens. Eight antigens were purified fractions from tumor membrane preparation, and five were crude membrane preparations from benign and malignant breast cell lines. Additionally, the Thomsen-Friedenreich antigen of erythrocytes was tested. We selected 83 hybridomas for further characterization.

Lung tumor-associated antigens: thin layer immunoassay.

Six hundred eighty-four rapid, sensitive thin layer immunoassays (TLI) were performed using an adapted visualization of condensation on plastic surface (VCS) method, for testing squamous cell and adenocarcinoma monospecific (TAA) and monoclonal antibody derived lung tumor-associated antigens (TAA epitopes) against small amounts of coded human test sera. The TAA epitopes selected for VCS-TLI were more specific, but both forms of TAA gave positive tests in 94% of lung cancer test sera of the appropriate histological types. The four antigens (adeno TAA and TAA epitope, squamous TAA and TAA epitope) did not react in VCS-TLI with coded sera from preselected normal individuals with known medical histories. With both TAA epitopes, 5% of non-lung cancer test sera were positive. There were indications that such tests may be useful in aiding pathological evaluations and in detecting precancerous conditions in patients with asbestosis. Cross-reacting (with other non-lung cancers) and specific peptide sequences of TAA were indicated in the comparative tests with the corresponding TAA epitopes, suggesting the tests for probing and analyzing portions of the antigens. Of immediate use is VCS-TLI for monitoring patients on specific active TAA immunotherapy.

Specific and nonspecific immunotherapy as an adjunct to curative surgery for cancer of the lung.

Attempts to improve survival following curative surgery for non-small-cell lung cancer are reviewed. Most of these approaches have been designed to stimulate the resistance of lung cancer patients in a non-specific fashion. Living bacteria or products of dead bacteria have been given as adjunctive treatment. Various routes have been used; oral, intradermal, subdermal, or intrapleural, with either BCG or Corynebacterium parvum. No reproducible benefit has been observed. Levamisole has not been proven to be useful. Trials have yet to be completed to confirm the use of thymosin fraction V for small cell carcinoma in improving the effectiveness of chemotherapy. A pilot trial using specific active immunotherapy is described. Prolongation of survival four years after closure of the trial in those patients immunized, compared with non-immunized patients, has prompted two further clinical trials. A small trial has confirmed the effectiveness of specific immunotherapy as adjunctive therapy for squamous cell carcinoma. A large multicenter trial in Canada and the United States should be completed and open to analysis in 1984 and may shed light on the role of tumor-associated antigens in stimulating specific resistance to lung cancer.

Partial purification of a common antigen in bovine lymphoma and its use in a lymphocyte blastogenesis assay.

An antigen was isolated from tumor cells derived from cases of bovine lymphoma which caused the in vitro blastogenesis of lymphocytes from nine of 13 (69%) adult cattle with lymphoma. Lymphocytes derived from five of 122 (4%) normal adult cattle also underwent blastogenesis while lymphocytes from three cases of the sporadic form of lymphoma did not respond. Blastogenesis of lymphocytes from normal cattle may have resulted from alloantigen activity in the antigen; however, normal lymph node antigen did not stimulate lymphocytes from normal adult cows or cows with lymphoma. Antigen derived from tumor was localized to the cell membrane and cytoplasm of tumor cells by indirect immunofluorescent staining using an antiserum prepared in rabbits. Specific fluorescence was reduced by the absorption of this antiserum with a crude tumor extract. The antiserum did not cross-react with normal lymph node antigen or viral components as demonstrated by immunodiffusion. With further refinement of the antigen, the blastogenesis assay may be of use in the early detection and study of the pathogenesis of lymphoma in adult cattle.

Cancer serum index: a useful nonspecific test as a parameter in multimodality screening and assessment of patients with cancer of the prostate.

A satisfactory, simple test for screening and monitoring of patients with prostatic cancer is still being sought. We suggest the use of a nonspecific test in combination with other tests. The ratio of two serum proteins (alpha 1 acid glycoprotein [AGP] and prealbumin [PAB]), may be useful markers for a cancer serum index (CSI). By separating serum components by gradient polyacrylamide gel electrophoresis, staining and quantitating these proteins by scanning densitometry, and dividing the area of AGP by that of PAB, indices can be obtained. In a test of sera from 450 patients with prostatic cancer, other urologic problems, and normal controls, CSIs of 360 sera presently decoded were found to be: (Formula: see text). The CSI should prove highly valuable when used in combination with selected prostatic cancer tests.

Treatment of inoperable lung carcinoma: a combined modality approach.

Twenty-four patients with inoperable lung carcinoma other than of the small cell type who received cis diamminedichloro platinum (II)-based combination chemotherapy were further treated with all available treatment modalities: radiation therapy, lung resection, chemotherapy, and immunotherapy. There were 2 operative deaths, and 2 patients died 6 and 8 months postoperatively of cardiac causes. Postmortem examination on these 4 patients revealed no evidence of residual tumor. The remaining 20 patients are alive 7 to 33 months from the onset of chemotherapy and 4 to 27 months following lung resection. These results, although preliminary, are encouraging, and further study is in progress.

Evaluation of the relationships of prealbumin components in sera of patients with cancer.

Serum proteins of cancer and control sera have been evaluated by the conventional Mancini method as well as by scanning densitometry measurements of components after separation by gradient PAGE. We established than in sera of cancer patients, prealbumin levels decrease and alpha 1-acid glycoprotein levels increase significantly. However, the Mancini method was not as sensitive as PAGE for quantitative evaluation of differences among sera of healthy adults, smokers, and patients with benign disease. We find that a more sensitive measure is the ratio between the two components (alpha 1-acid glycoprotein and prealbumin) and suggest that these values possibly may be useful as a CSI. In this study, the average CSI for healthy adults is 0.79, for heavy smokers, 1.24; for benign diseases, 1.67; and patients with various forms of cancer as follows: early lung, 4.17; late lung, 8.78; bladder, 8.92; colon, 13.77; breast, 2.43; and melanoma, 9.22.

Skin tests with soluble tumor membrane antigens in patients with transitional cell cancers.

Patients with transitional cell cancers and control patients with other forms of cancer were tested with cell membranes and soluble membrane antigens with the use of delayed hypersensitivity skin tests. These ongoing and parallel studies in which LMI tests are used have not been completed.

Preliminary studies of sera and urine from patients with bladder cancer.

Some preliminary observations may be helpful as we proceed with studies of the humoral and cellular reactions in body fluids. 1) In the reactions with TAA from primary bladder cancers and from bladder cancer cell lines, we must be aware of complexing when we study sera or tumors from patients with more invasive tumors. 2) Earlier stage patient sera can react to antigens present in urine but may not react with antigens present on tumor cells. These reactions are mainly related to cell sediments or partially insoluble material present in the urine; it is difficult to interpret these reactions in a controlled study. 3) Using sera from patients with different stages of bladder cancer, we can test normal bladder cell lines (e.g., HCV-29, which has no CF reactivity with sera from patients with bladder cancer) for comparison with the patterns of TAA present in cancer cell lines (e.g., RT-4 and T-24). 4) There is no relationship between HSV and bladder TCC. 5) Cytomegalovirus and its components are associated with normal or cancer-related cell and urine components. We can study differentiation of such antigens from TAA, which produce cell-mediated immune responses.