Variables and specificity of in vitro lymphocyte-mediated cytotoxicity in human melanoma.

In vitro cell-mediated cytotoxicity (CMC) assays have been carried out in human melanoma system with blood effector lymphocytes on [3H]proline-labeled target cells in a 48-hr microcytotoxicity technique. Three lymphocyte purification procedures (Ficoll:Hypaque gradient, plasma gel sedimentation followed by nylon column incubation, and plasma gel sedimentation followed by separation with nylon powder and glass beads) are compared in parallel experiments for characteristic effector cell composition and cytotoxic potential against target cells of dissimilar histology. The cytotoxicity is defined by the loss of target cell 3H cpm as measured by residual target cell 3H cpm in individual microwell following incubation with lymphocytes. Target cell 3H cpm loss by test lymphocytes is compared with target cell 3H cpm loss by several age and sex matched control lymphocytes (from normal donors and unrelated cancer patients); further comparison between the various control lymphocytes is also made in each assay. As control for target cells, autologous fibroblasts and homologous tumor cells of dissimilar histology are always included in each assay. Specific cytotoxicity is defined as statistically significant and selective destruction of only melanoma cells by the test lymphocytes as compared to the control lymphocytes. Significant but nonselective destruction of 2 or more target cells of unrelated histology is regarded as nonspecific cytotoxicity, while no destruction of any target cells signifies no cytotoxicity. The Ficoll:Hypaque preparations consistently exhibit the highest nonlymphocytic cell contamination (8 to 16%); the nonlymphocytic cells are, almost exclusively, monocytes. They also produce relatively high percentage of thymus independent (B) cells (8 to 15%). The ultimate cell composition of the 2 plasma gel-nylon preparations is essentially identical. In either plasma gel-nylon preparations, the nonlymphocytic contamination is minimal (0 to 4%) and thymus-dependent (T) cell percentage is considerably higher (92 to 99%) with none or few B cells.

Emergency and urgent operations for ulcerative colitis. The procedure of choice.

Eighty consecutive emergency and urgent colectomies for ulcerative colitis were performed. One-stage total proctocolectomy was performed in 37 patients, with a 9.1% mortality; ileostomy with subtotal colectomy was performed in 43, with a 7.0% mortality. The overall mortality was 7.5%. Postoperative morbidity after total proctocolectomy (mean postoperative hospitalization, 27.6 days; nonseptic complication rate, 29.4%; septic complication rate, 29.4%) was not substantially different from that after subtotal colectomy (postoperative hospitalization, 33.3 days; nonseptic complications, 45.0%; septic complications, 35.0%). Survivors of subtotal colectomy required abdominal-perineal resection of the colorectal remnant in 75.7% of patients, and no patient had successful subsequent ileorectal anastomosis. It is suggested that one-stage total proctocolectomy be adopted as the surgical procedure of choice in emergency or urgent operations for ulcerative colitis.

Fate of the retained rectum after subtotal colectomy for inflammatory disease of the colon.

Of forty-nine consecutive patients who underwent subtotal colectomy for inflammatory disease of the colon, 73.5 per cent required subsequent combined abdominoperineal resection of the retained rectum and only two patients had successful ileoproctostomy. For the patient whose rectum is substantially diseased along with the rest of the colon, one-stage total proctocolectomy is the preferred operation.

Staging laparotomy in Hodgkin's disease.

In a series of 34 consecutive staging laparotomies for Hodgkin's disease, unsuspected disease was discovered below the diaphragm in four cases (11,8%). The clinical staging was changed in 11 cases (32,4%). These data correspond to the results in most large series. Since staging laparotomy can be performed with minimal morbidity and gives the clinician the information essential to a rational plan of therapy, it is suggested that staging be performed prior to initiation of treatment in all patients with Hodgkin's disease not clinically proven to be disseminated.

The benefits of surgery for asymptomatic gallstones.

The risk of a patient developing complications of asymptomatic cholelithiasis is high enough to justify the mortality associated with prophylactic cholecystectomy. The operation effectively stops the progression of calculous biliary tract disease. If performed when the patient is young enough and healthy enough to tolerate the operation, and before the occurrence of complications which increase the mortality risk, the overall mortality and morbidity of calculous biliary tract disease could be significantly reduced.

Retroperitoneal liposarcoma.

A group of patients with retroperitoneal liposarcoma is presented and general guidelines of management reviewed. It is suggested that aggressive, radical total surgical excision followed by adjuvant postoperative irradiation be used as the treatment of choice. Although the management of recurrent disease may be discouraging in terms of ultimate cure, reasonable palliation and prolonged survival may result from aggressive therapy.

Heterotopic pancreas: gastric outlet obstruction secondary to pancreatitis and pancreatic pseudocyst.

Heterotopic pancreas, usually a silent gastrointestinal anomaly, may become clinically evident when complicated by a pathologic process. We report a unique case in which pancreatitis and pseudocyst formation in an antral lesion produced gastric outlet obstruction. The nature of heterotopic pancreas, its diagnosis, and management are discussed.

Selective and non-selective lymphocytotoxicity in human melanoma: observation on the effect of long-term culture and fetal bovine serum on target-cell sensitivity to lymphocytes.

In vitro cell mediated cytotoxicity (CMC) assays have been conducted in a human melanoma system with a 3H-proline retention technique. Melanoma target cells from long-term cultures ("cell lines") are found to exhibit increased susceptibility for lymphocyte cytotoxicity in comparison to the same target cells from short-term culture. The higher sensitivity of the "cell line" derived target cells is seen with lymphocytes, irrespective of diagnosis of the donor. In parallel experiments with the target cells grown in medium supplemented with fetal calf serum (FCS) and AB+ human serum (from a normal male doner), the melanoma target cells grown with FCS do not show any enhanced cytotoxicity, suggesting no causal relationship of such enhanced sensitivity of "cell line"-derived target cells to "heterologous melanoma antigens" that might have been acquired by the target cells following the use of FCS in tissue culture. In controlled assays of in vitro CMC, lymphocytes from melanoma patients (14/44) exhibited selective cytotoxicity (destruction of only one target-cell type) against the melanoma target cells, whereas only 3/97 control lymphocytes (other malignancies and normal donors) showed such melanoma-selective cytotoxicity. This difference is statistically significant at p less than 0.001. Non-selective cytotoxicity (destruction of two or more unrelated target cell types) was seen with lymphocytes from 9/44 melanoma patients, 13/51 patients with other malignancies and 8/46 normal donors. No correlation of selective cytotoxicity could be established with donors' age, sex, stage of disease, therapy or history of blood transfusion. Such a correlation may emerge as our series becomes larger. Despite the lack of any correlation between selective cytotoxicity and disease status, our study reaffirms the existence of selective cytotoxicity by melanoma patients' lymphocytes against melanoma target cells.