Ariel C. Hollinshead

Antibodies to Herpesvirus Nonvirion Antigens in Squamous Carcinomas

Serums from tumor-bearing patients, cured patients, and normal subjects were examined for antibodies to the separated complement-fixing reactive components of nonvirion antigens of herpesvirus type 1 and type 2. The occurrence of antibodies to the antigens was similar in serums from tumor-bearing patients and cured patients. Antibodies to the antigens were observed among 21 of 24 (87 percent) cervical cancer cases, 44 of 49 (90 percent) laryngeal cancer cases, 15 of 24 (62 percent) cases of squamous cell carcinomas of the head and neck excluding the larynx, 2 of 24 (8 percent) nonsquamous cell cancer cases, and 3 of 51 (6 percent) normal subjects. By contrast, no differences were found in the titers of neutralizing antibodies to the virus in serums from laryngeal cancer patients and controls. The observations support an etiologic role of herpesviruses in cervical cancer and in laryngeal cancer, and possibly other squamous cell cancers of the head and neck.

Cutaneous Delayed Hypersensitivity Reactions to Soluble Melanoma Antigen in Patients with Ocular Malignant Melanoma

Abstract We studied the cutaneous delayed hypersensitivity responses to a soluble melanoma antigen in 32 patients with the initial clinical diagnosis of ocular melanoma and in seven control patients. Eighteen out of 19 patients who had pathologically confirmed ocular melanomas were positive to this antigen, as were eight other patients clinically thought to have choroidal melanomas. All seven controls were negative on skin testing with this antigen, as were five patients who were initially thought to have ocular melanomas but who, on extensive work-up, were considered to have other, nonmelanoma, ocular lesions. Thus, patients with ocular malignant melanomas have cell-mediated immunity against an antigen common to systemic malignant melanoma. This delayed hypersensitivity assay may assist in the diagnosis of ocular melanoma, especially in patients with opaque media or those in whom ocular melanoma must be distinguished from metastatic lesions to the choroid. (N Engl J Med 291:274–277, 1974)

IMMUNOCHEMOTHERAPY OF LUNG CANCER

After surgical resection of their primary lung cancer, 33 patients were randomized into one of three groups. The first received high-dose methotrexate once per month with citrovorum rescue, for 3 months. The second group were immunized monthly with a homogenate of Freunds complete adjuvant and carefully characterized soluble antigen derived from allogeneic lung cancer cells of appropriate histology, for 3 months. The third group received a combination of methotrexate and immunization monthly, for 3 months. Each patient was monitored immunologically before, during, and after the treatment period, by use of delayed hypersensitivity reactions to recall and cancer antigens, in vitro lymphocyte response to mitogens, and mixed lymphocyte blocking factor activity. The group that received methotrexate showed little change in skin reactivity, a reduction of blocking factor activity, and significant rebound overshoot in in vitro lymphocyte performance. The immunized group showed a tendency to production of blocking factor activity, striking conversion and enhancement of skin reactivity, and little change in in vitro lymphocyte performance. The immunochemotherapy group showed dramatic increases in specific skin reactivity to cancer antigens, up to 2 years after treatment, in vitro lymphocyte rebound overshoot, and reduction of blocking factor activity production. Classic life table analysis of the probability of freedom from metastases in patients with stage-I cancer indicate that the disease-free interval in patients who received methotrexate is longer than in historic and concomitant controls but not as long as in those who received immunization. The best group appear to be those who received combination immunochemotherapy. We emphasize that the small numbers in this pilot study do not yet allow firm conclusions to be made.

Analysis of soluble melanoma cell membrane antigens in metastatic cells of various organs and further studies of antigens present in primary melanoma.

In malignant melanoma, using Sephadex G‐200 chromatography and polyacrylamide gel electrophoresis (PAGE), it has been possible to separate two types of skin reactive antigens. The first, found in Sephadex fraction II and PAGE region a appears specific for melanoma. Allogeneic extracts have produced positive reactions in many patients with skin or ocular melanoma, and have given negative reactions in patients with other types of cancer or in patients with ocular lesions simulating melanoma. The second group of antigens, in Sephadex fraction III and PAGE region b were less specific. These antigens produced positive skin reactions in some patients with breast cancer, as well as in patients with melanoma. Reactivity to PAGE region a appeared to be confined to one protein band, but three different bands in region b gave positive reactions. A study was made of the presence or absence of similar antigens in metastatic deposits of malignant melanoma. Metastatic lesions in the following tissues were analyzed: liver, lung, adrenal, skin, and colon. These were compared with pooled primary skin melanomas by skin testing in the same patients. The tumor‐associated melanoma antigen, found in Sephadex fraction II and PAGE region a appeared to be strongest in adrenal, lung, and liver metastases. It was found that the protein yield in this region was not indicative of the strength of the antigen. Therefore, a careful, detailed analysis of the protein bands present in PAGE regions a and b from primary skin melanoma was conducted. Only one band in PAGE region a was found to be responsible for positive skin reactivity. This band was found to be a glycolipoprotein. Further studies were also conducted in order to determine whether or not some of the antigens present might be fetal antigens. Some of the protein bands present in Sephadex fraction III and PAGE region b of melanoma appeared to be similar to some of the PAGE region b proteins present in fetal skin cells. Two bands from fetal skin also had the same location on PAGE as two bands from ductal breast cancer, although the relationship to melanoma region b antigens was not exact. These fetal proteins, which seemed to be present both in ductal breast cancer cell membranes and in melanoma cell membranes, might account for the positive skin reactivity seen in this region, and also for the cross reactivity of skin tests with this antigen.

Familial occurrence of colon and uterine carcinoma and of lymphoproliferative malignancies. II. Chromosomal and immunologic abnormalities.

Cytogenic and immunologic studies were performed on 20 members of a family who had an increased susceptibility to carcinomas of the colon, uterus and of lymphoproliferative malignancy. Chromosomal abnormalities such as small G and/or long submetacentric marker chromosome and other aberrations were observed in members who had cancer as well as some asymptomatic siblings. In addition, impairment of T cell function was noted in many of the members suggesting that defective cell‐mediated immunity, which also may be genetically determined, played an important role in the expression of this disease.

Identification of a Soluble Transplantation Antigen from the Membrane Fraction of Adenovirus Tumour Cells

Summary A soluble fraction containing transplantation antigens was isolated from cells infected with adenoviruses and from the membrane fraction from cells of adenovirus-induced hamster tumours. It was not identified either as a part of the virus structure or as a T antigen. This material regularly protected young adult hamsters against the development of tumours resulting from the transplantation of adenovirus tumour cells. Protection was given by soluble antigens isolated from more than one cell type; the antigens could be induced by at least two groups of adenovirus, with probable cross-protection within groups. The antigens did not protect animals challenged with tumour cells produced by a different DNA virus, SV40.

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.

Detection of herpes simplex virus tumor-associated antigen in uterine cervical cancer tissue: five case studies.

Abstract Biopsies from five gynecologic cancer patients were examined for the presence of herpes simplex virus tumor-associated antigen (HSV-TAA) using the highly sensitive peroxidase-antiperoxidase technique. The results demonstrate that two of three patients with squamous cell carcinoma of the uterine cervix that were serologically positive for HSV-TAA, determined by complement fixation, contained HSV-TAA in the cytoplasm of the malignant cells of the biopsy. Tissue from one squamous cell carcinoma of the uterine cervix patient, as well as tissue from an adenoacanthoma patient, was negative for tissue HSV-TAA. However, both of these patients were also serologically negative for HSV-TAA. All patients, regardless of cancer type, had serum antibodies to herpes simplex virus types 1 and 2. These studies continue to suggest a role for HSV in certain gynecologic cancers.

Skin tests with soluble melanoma antigens in patients with choroidal tumors

The cutaneous delayed hypersensitivity reactions with a soluble melanoma antigen in patients with choroidal mass lesions were studied. Ninety percent of patients with pathologically documented choroidal melanomas had positive melanoma antigen skin test responses. There did not appear to be any difference in the histologic appearance of the tumor nor in the disease status of those patients with positive versus those patients with negative skin test reactions. A 21% instance of false‐positive responses with this partially purified soluble melanoma antigen in patients with simulating lesions was observed. The cause for this reactivity is unclear; however, from previous work tissue‐associated antigens or fetal antigens are the most probable etiologies for the false‐positive melanoma antigen skin tests observed. Further purification of the melanoma‐associated antigen preparation may increase the specificity. The results of this study would mitigate against the use of this soluble melanoma antigen skin test in the primary evaluation of patients with pigmented choroidal mass lesions. Currently, the assay is being tested to ascertain its correlation with prognosis and as a means of monitoring immunotherapy.

Localization of complement-fixing antigens in cells: Epstein-Barr virus-induced membrane and interior cell antigens.

Summary A direct study of the complement-fixing reactivity of cell membranes, soluble components of cell membranes and cell interiors of lymphoblast cell lines showed that with highly concentrated levels of antigen, complement-fixing reactivity could be measured consistently between the cell interior of lymphoma (IM1), leukaemic (4265) and Burkitt lymphoma (P3) cultured cells and sera from patients with infectious mononucleosis or Burkitt lymphoma. A measurable complement-fixing reactive cell-membrane antigen appeared to be present in IM1 and 4265 cells. Complement-fixing activity was also recovered in a soluble fraction of 4265 cell membranes after sonication and separation by Sephadex gel filtration. Separation of the complement-fixing reactive component from HL-A antigens was achieved by gel electrophoresis.