Description of a murine model of experimental hepatic metastases.

A murine model of experimental hepatic metastases has been developed. The cecum is exteriorized through a midline incision, and 1.5 X 10(5) MCA-38 liver-derived (LD) tumor cells in 0.1 ml was injected into the ileocolic vein (ICV). Ninety-eight percent of injected mice developed hepatic foci. The operative mortality was 6.1%. Micrometastases could first be detected on day 11. Laparotomy of 21 days revealed the presence of a mean of 18 hepatic foci. Experimental hepatic metastases could be palpated 35 days following ICV injection. Mice bearing MCA-38 LD foci survived an average of 53.3 days. This model will be of use in the development of novel approaches for the treatment of hepatic metastasis.

Murine colon adenocarcinomas: methods for selective culture in vitro.

Two murine colon adenocarcinoma cell lines were established from primary cultures. The MCA-38 cell line was begun by treatment of the primary culture with trypsin to remove the fibroblastoid elements. The MCA-36 epithelial cells were sensitive to trypsin; therefore, the growth medium of MCA-36 primary cultures was augmented with collagenase to release the tumor-cell elements from the fibroblast network. These tumor elements were dissociated with trypsin and placed in tissue culture. Each cell line was cultured for at least 10 passages in vitro and gave rise to tumors when reimplanted in vivo.

Murine colon adenocarcinoma: syngeneic orthotopic transplantation and subsequent hepatic metastases.

Syngeneic murine colon adenocarcinoma (MCA-38) cells were transplanted in the submucosa of distal colon, proximal colon, cecum, ileum, jejunum, and duodenum of male C57BL/6 mice, with local lymphoid follicles used as points of entry. The tumor grew best at the cecum and led to liver and mesenteric lymph node metastases in 8 and 9 weeks, respectively, after transplantation. Histologically, a local inflammatory reaction involving polymorphonuclear leukocytes was observed within 48-72 hours following transplantation; after this time, the microscopic tumor foci began to grow progressively. Mononuclear lymphoid cells of the gut-associated lymphoid tissue did not infiltrate the progressively growing tumor; however, polymorphonuclear leukocytes were constantly observed at the tumor periphery in the lamina propria. The studies indicated that orthotopic transplantation as a model system can provide a means of examining the role of the local immune response as a focus of host resistance and as a factor in metastatic tumor spread. The findings also suggested the usefulness of this model in immunotherapeutic and chemotherapeutic studies of secondary hepatic disease.

Identification of a macromolecule containing an anticarcinoembryonic antigen-reactive substance and immunoglobulin M in human pancreatic cancer.

Ascitic fluid from a patient with carcinoma of the pancreas was fractionated by ammonium sulfate precipitation. The fraction precipitated between 25 and 50% saturation of ammonium sulfate was sequentially chromatographed on Sephadex G-200 and Sepharose 6B. A macromolecular fraction (greater than 10(6) daltons) obtained was found to react with both antihuman IgM and antiserum to carcinoembryonic antigen (CEA). This fraction was further purified by adsorption with protein A-Sepharose CL-4B and chromatography on DEAE-Sephacel. The purified macromolecular fraction had a sedimentation value of 28S as determined by ultracentrifugation. Upon dissociation of the purified macromolecule at pH 2.3 and purification of the dissociated components on Sepharose CL-2B and BioGel A 1.5M, a 19S protein and a 5S protein were recovered. The 19S protein showed a complete line of identity with a reference human IgM when reacted with antihuman IgM in gel diffusion, whereas the 5S protein showed a partial immunologic identity with colon CEA against anti-CEA. These results indicated the existence of an IgM-containing macromolecular complex with an anti-CEA cross-reactive substance in the extracellular fluid of human pancreatic cancer.

Demonstration of the microtest version of the leukocyte adherence inhibition assay.

This paper gives a detailed description of the microtest leukocyte adherence inhibition technique, as well as the results obtained with blood specimens coded by impartial observers. Three coded blood specimens from patients with colon cancer, pancreatic cancer, and melanoma were tested against crude membrane preparations of pancreatic and colon adenocarcinoma tumors. No tissue type-specific reactivity was observed. The inability to demonstrate specific reactivity was due to extensive variability observed within each test. The extensive variability resulted from time constraints of the workshop that necessitated deviations from the normal procedure.

Isolation of glycoprotein antigen from ascites fluid of pancreatic carcinoma.

A glycoprotein antigen from the ascites fluid of pancreatic carcinoma has been isolated with the use of perchloric acid extraction and chromatographies on Sepharose 4B, DEAE-Sephadex, and DE52, followed by Sephadex G-200. The purified glycoprotein was found to be homogenous by sodium dodecyl sulfate polyacrylamide gel electrophoresis. A molecular weight of 185,000 dattons was determined by gel filtration. The molecule contained 55% protein and 45% carbohydrate. Both the amino acid and carbohydrate compositions were different from those of carcinoembryonic antigen. This glycoprotein antigen of pancreatic cancer cross-reacted with anticarcinoembryonic antigen; however, the antiserum prepared from this antigen did not react with carcinoembryonic antigen. The biological significance of this glycoprotein in pancreatic cancer is being studied.

Cellular and humoral factors involved in the mechanism of the micro-leukocyte adherence inhibition reaction.

To study the cellular basis for specific antigen-induced leukocyte adherence inhibition, enriched populations of B-cells, T-cells, and monocytes were prepared by a two-stage adherence separation procedure from spleen cells of normal C57BL/6J mice and mice bearing progressively growing MCA-38 tumors. The reactor cell undergoing specific antigen-induced adherence inhibition was identified as a monocyte (esterase positive, did not respond to mitogens, and did not bear Thy 1.2 antigen or surface immunoglobulin). Furthermore, an enriched population of MCA-38-sensitized B-cells could program normal monocytes to undergo specific antigen-induced adherence inhibition. In contrast, enriched populations of MCA-38-sensitized T-cells could not program normal nylon wool-adherent cells to undergo antigen-specific adherence inhibition. Programming of normal monocytes by MCA-38-sensitized B-cells occurs through a soluble mediator and not by direct cell contact. The soluble mediator appears to be immunoglobulin in nature and induced both adherence inhibition and the inhibition of adherence. Thus, in this murine tumor model, leukocyte adherence inhibition appears to be due to programming of monocytes by a secretory product of specifically sensitized B-cells.

Correlations between the leukocyte adherence inhibition microassay and in vivo tests of transplantation resistance.

The leukocyte adherence inhibition (LAI) microassay detects tumor-associated antigen(s). Extracts of colon carcinoma (MCA-38 and B16 melanoma tumors, both syngeneic to the C57BL/6J mice) are recognized only by peritoneal cells from mice bearing the corresponding tumor. To ascertain whether this in vitro antigenic recognition correlates with the ability of the host to recognize and reject a tumor in vivo, serial LAI microassays were performed synchronously with experiments designed to test the ability of mice bearing tumors to reject live secondary tumor challenges. Concomitant tumor immunity was present in the MCA-38 tumor-bearing mice on 3 occasions from 5 to 15 days from primary inoculation. In the B16 system, concomitant immunity was present on one occasion 10 days after primary inoculation. These results in turn were paralleled with the specific in vitro recognition of tumor antigens as detected by the LAI microassays. Loss of immunity in the "eclipse" phase of tumor development, as detected by concomitant tumor immunity, was paralleled by nonreactivity of the indicator cells in the LAI microassay.

Semi-automated method for the enumeration of cytotoxicity results.

This communication provides information on the design and use of a counting machine that enumerates adherent cells in cytotoxicity assays. To date, investigators have relied on inverted microscopes or quantimat automatic image analysis. The former method is tedious and time consuming while the latter method is prohibitively expensive. The counting machine which consists of projection microscope attached to an electronic counter is as accurate as the inverted microscope but is both less tedious and less time consuming.

Isolation of human peripheral blood lymphocytes: modification of a double discontinuous density gradient of Ficoll-Hypaque.

Incubation of whole blood with adenosine diphosphate (ADP) and carbonyl iron prior to layering on a double discontinuous gradient of Ficoll-Hypaque selectively reduced the number of platelets and monocytes found at the upper lymphocyte interface after centrifugation. This modification in combination with a double discontinuous gradient results in a lymphocyte isolation method that is simple, rapid and reproducible.

Leukocyte adherence inhibition: an automated microassay demonstrating specific antigen recognition and blocking activity in two murine tumor systems.

This communication describes an automated micro-adherence inhibition assay. Tumor-specific immunity was demonstrated with B16 melanoma and MCA-38 colon adenocarcinoma, both of which are syngeneic to the same strain of mouse (C57B16/J). Abrogation of the leukocyte adherence inhibition (LAI) response of sensitized leukocytes has been demonstrated in the MCA-38 tumor system by the addition of serum from mice bearing MCA-38 progressively growing tumors, a property not present in normal serum. The sensitivity of the system has also been demonstrated by showing that LAI will change prior to a tumor becoming palpable. This microassay has the advantage of being simple, rapid and reproducible, and involves the use of minimal quantities of antigenic preparations and leukocytes, and in addition is amenable to rigorous statistical analysis.

CEA (carcinoembryonic antigen): its role as a marker in the management of cancer.

A consensus Development Conference was held at the National Institutes of Health from September 29-October 1, 1980, to address issues concerning the role of carcinoembryonic antigen (CEA) as a marker in the management of cancer. The panel met following formal presentations and discussions to assess the issues based on the evidence presented. These issues included: Should CEA be used in cancer screening? Is CEA helpful in cancer diagnosis? What does CEA tell about the extent and outcome of cancer? Is CEA helpful in monitoring cancer treatment? This paper constitutes the panel's findings.

Evaluation of palliative resection in advanced carcinoma of the stomach.

A retrospective study of 140 patients with adenocarcinoma of the stomach seen at Roswell Park Memorial Institute during a 5 year period revealed that 112 patients were TNM Stages III or IV. A palliative resection of the stomach was done in 39 patients; the other 73 did not undergo resection. Patients who had palliative resection for stages III and IV carcinoma of the stomach had a prolonged mean survival rate, as compared with those with similar stage of disease who did not have palliative resection, regardless of an additional treatment such as feeding procedures, gastroenterostomy, chemotherapy, and radiation therapy. This significance was noticed regardless of age, sex, and histological differentiation of the tumor. Chemotherapy significantly increased the survival rate in the unresected group but failed to show this effect in the resected group.

Survival of patients with stage B2 colon carcinoma. The Gastrointestinal Tumor Study Group experience.

The Gastrointestinal Tumor Study Group, a multispecialty collaborative clinical research program, executed two studies of surgically resected colon carcinoma, including patients with serosal penetration but no positive regional lymph nodes (stage B2). In the first study, 232 patients with stage B2 disease were operated on and survival was unaffected by therapy. In the stage B2 group, survival was 78% at five years, and is estimated to be 64% ten years after surgery. A second study, begun in 1979, compared survival and recurrence data on patients who underwent surgical resection and were treated postoperatively with fluorouracil and hepatic irradiation vs those who were observed. Hematologic toxicity of the regimen and accumulation of data from the first study caused the termination of randomization of patients with stage B2 disease with continued registration to observation only. One hundred twenty-four untreated patients with stage B2 carcinoma had a five-year survival rate of 77%, confirming the relatively high survival of this group of patients who are treated with surgery alone. Our patients' five-year survival of 78% corresponds favorably to the 87% survival rate anticipated in an age-, sex-, and race-matched cohort of Americans not specifically diagnosed as having cancer. Adjuvant trials, in seeking improved survival, should recognize that our review of 356 patients implies a considerably higher-than-anticipated five-year survival for the patient with stage B2 disease who undergoes surgical resection.(ABSTRACT TRUNCATED AT 250 WORDS)

Hemipelvectomy.

Hemipelvectomy was performed in 50 patients with malignant neoplasms of the upper part of the thigh and pelvis. Although not technically difficult, the operation is associated with considerable blood loss. Postoperative complications are frequent, the greatest morbidity resulting from skin flap necrosis. Symptomatic phantom limb is as occasional late problem. Of 37 patients resected for cure prior to 1969, 14 were alive five or more years postoperatively. Five of these long-term survivors subsequently died of metastases. Patients with fibrosarcoma and chondrosarcoma had the best survival. Six other patients underwent palliative hemipelvectomy for intractable pain, with gratifying results. Hemipelvectomy is an important, useful operative procedure in selected patients.

The surgical treatment and outcome of soft-tissue sarcoma.

Cases of 251 patients with soft-tissue sarcoma of the extremities, trunk, head and neck, and retroperitoneum were reviewed. The overall five-, ten-, and 15-year survival rates were 45%, 30%, and 24%, respectively. The five-year survival rates after wide soft-tissue resections, local excision, and amputation were 59%, 42%, and 39%, respectively. Factors that significantly affected prognosis were tumor size, histologic type, local extent of the tumor, and the adequacy of the surgical procedure. The local recurrence rate was 36% after wide resection, 8% after amputation, and 65% after local excision. Local recurrence alone was treatable and evidenced overall five- and ten-year survival rates of 43% and 33%, respectively. Amputation was the most effective method to treat local recurrences in the extremities. Some guidelines are suggested to select high-risk patients for adjuvant treatment.

BCG immunotherapy in patients with malignant melanoma.

Twenty-one patients with malignant melanoma received immunotherapy with BCG. Thirteen patients had adjuvant immunotherapy on a monthly schedule. Of these, eight with regional lymph node metastases (stage III) had been treated by lymphadenectomy. Two of the stage III patients had tumor recurrences within one year, while six are alive and free of melanoma at a median interval of 22 months. The remaining five patients (stage I or II) had level 4 or 5 (Clark classification) primary lesions. Their average tumor-free survival has been 18 months, but there was one regional recurrence in six months. Eight patients received intralesional treatment with BCG. The extent of local response correlated inversely to the stage of their disease. Higher doses of BCG or multiple simultaneous injections into the same lesion did not produce complete resolution of nodules in patients with far-advanced melanoma. In none was the course altered by intralesional therapy.

Endocrine organ metastases in subjects with lobular carcinoma of the breast.

OBJECTIVE: To define the frequency and pattern of endocrine organ metastases in patients dying of invasive lobular carcinoma. DESIGN: Postmortem microscopic evaluation of the ovaries and adrenal, pituitary, thyroid, and parathyroid glands for breast cancer metastases. SETTING: Roswell Park Cancer Institute, Buffalo, NY, between 1971 and 1990. SUBJECTS: One hundred eighteen subjects who died of their cancer: 86 had infiltrating ductal carcinoma; 32, invasive lobular carcinoma. MEAN OUTCOME MEASURE: Quantitative measurements to allow frequency determinations and statistical comparisons. RESULTS: Endocrine organ metastases were found in 91% of the subjects with invasive lobular carcinoma vs 58% of subjects with infiltrating ductal carcinoma. The adrenal gland was most frequently involved. Multiple endocrine metastases were most common in the group with invasive lobular carcinoma. CONCLUSIONS: A relationship exists between invasive lobular carcinoma and endocrine metastases. This indicates that antemortem endocrine evaluation may subsequently improve quality-of-life treatment.

Technic of resection of mesenteric tumors.

In the resection of mesenteric tumors or primary malignant lesions of the small bowel, exposure of the superior mesenteric vessels is important. This is the necessary criterion of resectability and the condition of obtaining maximal margins in the lymphatic drainage area. It also helps avoid unnecessary sacrifice of branches and tributaries to the superior mesenteric vessels from uninvolved portions of the bowel.

Intraluminal, lymph node, hepatic, and serum levels after intraluminal and intramural injection of 5-fluorouracil in the dog colon.

The tissue levels of 2-14C 5-fluorouracil were compared after three routes of administration in the colon of fifteen dogs. "Simple" submucosal injection resulted in higher levels in the colonic wall, abdominal lymph nodes, and liver than did rapid intravenous injection. Intraluminal administration demonstrated that the appearance of 5-FU was prompt and that significantly higher levels continued in the colonic wall and liver, but nodal levels were markedly lower.