D. M. P. Thomson

An overview: antitumor immunity in breast cancer assayed by tube leukocyte adherence inhibition.

The adherence to glass of human peripheral blood leukocytes (PBL) incubated with tumor antigen in vitro, is specifically inhibited if the PBL are sensitized to the antigen. The presence of leukocyte adherence inhibition (LAI) to tumor extracts indicates the presence of systemic antitumor immunity. By the tube leukocyte adherence inhibition assay (tube LAI), it was shown that 85% (191 of 223) Stage land II,45% (15 of 34) Stage III and 29% (30 of 103) Stage IV breast cancer patients had LAI reactivity. LAI responsiveness diminished with an increased tumor burden and most patients with advanced cancer exhibited no LAI reactivity. When LAI reactivity was monitored for 1 to 6 months after surgery, 13 of 25 Stage I and II breast cancer patients were negative on the first repeat assay. In general, 7 months after mastectomy most patients clinically free of cancer showed no LAI reactivity. Of thirty‐five patients tested between 7 and 18 months after mastectomy, 6 were positive and 4 of the positives had local recurrence. The phenomenon of tube LAI appears to be mediated by mon‐ocytes armed with cytophilic antitumor antibody. The serum of patients whose leukocytes responded in the tube LAI assay had free cytophilic antitumor antibody that “armed” or sensitized normal leukocytes to respond in the LAI assay. Serum arming paralleled leukocyte reactivity before and after surgery. Patients with advanced cancer whose leukocytes failed to react in the LAI assay had serum blocking factors (excess tumor antigen) that abrogated the LAI reactivity of leukocytes from reactive patients.

Predictive value of tube leukocyte adherence inhibition (LAI) assay for breast, colorectal, stomach and pancreatic cancer

The tube LAI assay measures accurately antitumor immunity in patients with early cancer but fails to detect up to 75% of patients with advanced cancer due to excess circulating organ‐specific neoantigen (OSN). Substances such as prostaglandin E2 (PGE2) or aminophylline which increase intracellular nucleotides in leukocytes of patients with advanced cancer reversed this nonreactivity and greatly increased the sensitivity of the assay without any loss of specificity. Antitumor immunity can now be detected in advanced cancer, and a combination of the two assays gives prognostic potential to the assay: a positive test with PGE2 and negative test without indicates the patient has a large tumor burden. The specificity of the assay for each cancer was high and in most instances was < 95%. The PGE2 stimulated assay retained the high specificity. The sensitivity of the regular tube assay was often low, 33–56% because of the many advanced cancer patients tested, whereas the PGE2 stimulated assay showed almost a two‐fold increase in sensitivity, 67‐93%. The diagnostic value of the assay was estimated by calculating the predictive value for different prevalences of cancer. It was found that at low prevalances of cancer as found in the general population, the assay had a low diagnostic value since few patients with a positive test would have the cancer tested for. With prevalences of cancer of 5% or greater as might be found in a tertiary care clinical setting, the assay would seem to have diagnostic value since one half or more patients with a positive test would have the cancer tested for. Most false positives, but not all, are found in patients who have lesions that are often considered to increase their risk for cancer: severe dysplasia of the breast, colon adenomas, chronic atrophic gastritis and chronic pancreatitis, suggesting that the assay predicts oncogenesis.

The Diagnostic Value of CA 27-29, CA 15-3, Mucin-Like Carcinoma Antigen, Carcinoembryonic Antigen and CA 19-9 in Breast and Gastrointestinal Malignancies

In the past decade, considerable interest has arisen for defining the role of various tumor markers in the diagnosis of cancer. This cross-sectional study evaluates four breast cancer markers (CA 27-29, CA 15-3, MCA and CEA) and two gastrointestinal (GI) markers (CA 19-9 and CEA) in 213 patients. Receiver operating curves (ROC) revealed a sensitivity for the 90% specificity cutoff for breast cancers compared to breast benign diseases of 70% for CA 27-29, 67.5% for CA 15-3, 52.5% for MCA and 40% for CEA. When GI tumors were compared to benign GI disease, the sensitivity for 90% specificity was 40.3% for CEA and 32.3% for CA 19-9. Comparison of breast cancer and GI malignancies with other malignancies leads to a marked shift of the ROC curve to the right and loss of specificity. Late stage for all breast and GI tumor markers was found to be a predictor of high serum antigen level (p < 0.001). The presence of liver metastases in breast cancer was associated with abnormal levels of CA 27-29 (p = 0.028). Pancreas adenocarcinomas had a higher CA 19-9 antigen level (p < 0.001) than other GI malignancies. CA 27-29 appears to be at least as sensitive and specific as CA 15-3 in patients with breast cancer. None of the above markers retain their specificity when compared with a control group consisting of other malignancies.

Human Cancers Transplanted in Nude Mice Retain the Expression of Their Organ- Specific Neoantigens*

Human cancers transplanted to nude mice seem to retain most features of the original tumor. To determine if organ-specific neoantigens were still expressed after transplantation, 4 different human cancers, supplied and coded by independent investigators, were assayed in the tube LAI assay. The LAI phenomenon measures the change in adherence properties of leukocytes that have bound an antigen to which they are sensitized immunologically. Four of four transplanted human cancers-pancreas, lung, breast, and colon—stimulated an LAI+ response with leukocytes from patients with cancers of the same organ. The antitumor response was not as vigorous against a pool of transplanted tumor extracts as against those prepared from liver metastases. Nonetheless, the results add to the mounting evidence that human cancers express neoantigens to which most humans bearing cancers respond immunologically.

Localized unilateral periorbital edema induced by aspirin

BACKGROUND Aspirin intolerance manifested as bronchospasm or urticaria/angioedema has been observed since the beginning of this century. OBJECTIVE To report a novel case of intolerance to aspirin ingestion. METHODS Case report; routine skin testing; pulmonary function testing; aspirin challenge. RESULTS A 30-year-old man with a history of left ocular trauma at the age of 10 noted a 3-year history of left periorbital angioedema after aspirin but not other nonsteroidal anti-inflammatory drugs. Incremental oral aspirin challenge resulted in this unilateral symptomatology at a dose of 673 mg. CONCLUSION To the best of our knowledge, this is the first reported case of unilateral periorbital edema following aspirin ingestion.

Comparison by leukocyte adherence inhibition of human immune response to cancer-associated immunogens, Thomsen-Friedenreich (T) and Tn, myelin basic protein, and organ-specific cancer neoantigens

Peripheral blood leukocytes from cancer patients exhibit nonadherence to glass as an index of antigen recognition when incubated individually with four distinct, soluble tumor-related substances. Crude cancer extracts, purified antigens, T and Tn, myelin basic protein (MBP), and organ-specific cancer neoantigens (OSN), all elicited narrow dose-dependent leukocyte adherence inhibition (LAI) response curves. The present study focused on the reasons for the narrow antigen dose-LAI response relationship. Between 9 and 20 pmol of antigens elicited the maximum number of nonadherent leukocytes; cleavage products of T antigen and the nonapeptide (T18) of MBP required about a 10-fold increase in molar concentration for the same LAI response. When crude cancer extracts were combined with pure antigen or the pure antigens were combined at concentrations shown to give maximum LAI responses, the positive LAI responses were negated. The chemoattractant LTB4 at 10(-11) M triggered maximum LAI. But when MBP was added with the LTB4 at progressively increasing concentrations, there was dose-dependent negation of LAI. The magnitude of LAI depended on the total amount of mediator released rather than the rate of release. When leukocytes from cancer patients were incubated with optimum to high concentrations of MBP, the supernatants contained a mediator that gave similar bell-shaped dose-LAI responses on control leukocytes indicating that leukocytes from cancer patients react to a much broader range of antigen concentration than indicated by the LAI assay alone. High antigen dose negated LAI because of excess mediator production. Antigen-generated mediators had a biphasic effect inducing nonadherence and then adherence of leukocytes.

Isolation of HLA and tumor antigens by means of affinity chromatography employing anti-β2,-microglobulin (β2m) antiserum

A method for the isolation of HLA antigen molecules from normal and cancerous solid human tissue is described. The method employs anti‐β2‐microglobulin (β2m) antiserum coupled to Sepharose beads as an immunosorbent affinity medium. The anti‐β2m affinity chromatography procedure greatly purifies and selectively enriches HLA and any material that copurifies by affinity, with β2m and /or HLA molecules. The HLA isolated by this purification procedure was used to immunize rabbits. The antisera obtained were absorbed on β2m to remove all anti‐β2m antibody activity. The use of such anti‐HLA antisera in radioimmunoassays, immunoprecipitation studies, and F(ab′)2 blocking experiments demonstrated that these antisera are directed against a common HLA determinant present on the heavy (alloantigen‐bearing) chain of all HLA molecules. The use of an identical procedure employing human tumor tissues has resulted in the isolation of HLA‐like or HLA‐associated tumor‐specific antigens as demonstrated by the leukocyte adherence inhibition (LAI) assay.

Measurement by Leukocyte Adherence Inhibition of Autosensitization of Cancer Patients to Myelin Basic Protein

In vitro cell‐mediated immunity was assayed by leukocyte adherence inhibition (LAI) to determine the extent of autosensitization to myelin basic protein (MBP). Leukocytes from 123 cancer patients, 16 patients freed of cancer, 135 patients with benign disease, and 26 patients with destruction of nervous parenchyma were tested. Most patients with cancer reacted to MBP: 92%, 93%, 82%, 78%, 75% and 62% for pancreatic, colonic, esophageal, lung, ovarian and breast. Few patients with benign diseases reacted to MBP. Patients with multiple sclerosis (MS) were sensitized to MBP, but patients with other nervous tissue injury did not react to MBP. Cancer patients did not remain sensitized to MBP once they were freed of their cancer. The LAI assay is a straightforward method of measuring cellular autosensitivity to MBP. In the population of patients tested, autosensitivity to MBP was confined, except for MS, principally to cancer patients.

Leukocyte activation in advanced cancer as an explanation for absent leukocyte adherence inhibition to cancer extracts and chemoattractant.

Tube leukocyte adherence inhibition (LAI) measures human immunity by antigen-binding leukocytes releasing chemoattractant-like mediators that are the ultimate inhibitors of adherence by bystander leukocytes. We determined whether the absent LAI responses to cancer extracts for patients with large body burdens of bladder cancer was related to a defect in antigen binding or chemoattractant responsiveness. Leukocytes from patients with small body burdens of bladder cancer gave positive LAI responses of a similar extent to either bladder cancer extracts or chemoattractant [n-formyl-L-methionylleucylphenylalanine (FMLP)]. Of the adherent leukocytes, about 20-30% became non-adherent with a positive LAI response: monocytes, neutrophils and lymphocytes responded. In the control tubes, leukocytes from patients with large body burdens of bladder cancer were more non-adherent and about 15% less adherent than leukocytes from controls or patients with early cancer. They showed no further decrease in adherence, or conversely increase in non-adherence, with either extracts of bladder cancer or FMLP. The leukocytes also failed to transduce transmembrane signals to the same stimuli. The defect was reversible since PGE2 restored the adherence of leukocytes to normal, and subsequently they exhibited membrane potential changes and about 34% non-adherence to either bladder cancer extracts or FMLP. From these results we conclude that chemoattractant LAI-responsive leukocytes from patients with large body burdens of bladder tumor are activated in vivo, probably by mediators from inflammatory cells.

Impairment of systemic concomitant antitumor immunity by excess soluble tumor antigen

SummaryAnimals bearing a 3-methylcholanthrene induced sarcoma called MC1 rejected substantial numbers of a suspension of the same tumor cells injected IV in comparison with normal rats. The factors that protected the host against lung metastases were impaired by the administration of tumor antigen in the form of irradiated tumor cells or soluble tumor antigen. Animals bearing an MC1 tumor which received either unrelated MC11 irradiated tumor cells or soluble tumor antigen had more lung metastasis than the animals not given any tumor products. However, a statistically significant increase in the number of lung tumor nodules was observed in the rats treated with MC1, compared with those treated with MC11 tumor antigen (soluble tumor antigen or irradiated tumor cells) or no tumor antigen. The increase in the outgrowth of lung tumor nodules in the tumor-bearing host given an excess of tumor materials was produced by a dual mechanism of inhibition of the concomitant immune resistance and nonspecific resistance. The present study shows that soluble tumor antigen similar to material shed from a primary tumor is able to impair concomitant immune resistance to tumor cells within the lungs.