Vaccinia melanoma oncolysates (VMO) were tested in a Southeastern Cancer Study Group (SECSG) Phase I/II trial. Forty-eight patients with high-risk Stage I or pathologic Stage II disease were placed on study at six different dose levels and two different treatment regimens. Patients were monitored for toxicity to the VMO after each injection. Patients' sera were tested for anti-human melanoma reactivity with the Staphylococcus Protein A (SpA) assay. Toxicity was minimal at all doses tested. In only 2 of 19 patients on delayed treatment did reactivity develop in the SpA assay by 6 months. However, 13 of 23 patients on immediate treatment showed reactivity, including 8 of 8 at the two highest doses. Since the VMO appears to be safe at all of the doses tested, and because of the immunogenicity of the VMO at the higher doses as demonstrated by the SpA assay, the 2-mg dose level, for immediate treatment, was chosen for use in future trials.
T memory cells specifically responsive to ovalbumin and performing the diverse functions of DNA synthesis, lymphotoxin release, and regulation can be isolated in enriched numbers in the most buoyant fractions (A+B) of bovine serum albumin gradients on slay 9 after sensitization. At least 20-30% of these cells are capable of mounting a blastogenic response to ovalbumin. A+B cells responding to ovalbumin with DNA This study is directed to three lymphocytic functions: the DNA synthetic response after restimulation, lymphotoxin production, and nonspecific suppression of the phytohemagglutinin (PHA) response by antigen. We have previously shown that in rats, after footpad injection with ovalbumin (OA) in complete adjuvant, the DNA synthetic response of draining lymph node cells on restimulation in vitro with OA peaks at day 9 (1). A peak in the ability of the same cells to form lymphotoxin in response to OA also is seen at day 9 (2). Finally, the nonspecific suppression of PHA responses by antigen (3) observable as early as 6-12 hr after a primary injection of OA (4) is strongly expressed at 9 days in adjuvant-injected animals and appears to be associated with a population of glass-wool-adherent cells (4,5).To further study T Separation of OA-Sensitized LNC. 1. Bovine serum albumin gradients: LNC were separated on discontinuous gradients (bovine serum albumin, lot 126, Miles Laboratories, Kankakee, Ill.) as described (1). Pools7 of cells harvested from the density interfaces and related interband regions were designated A, B, C, and D, beginning with the least dense band, and P (pellet) (Fig. 1). Viability of cells after washings was always >95%, excluding P (>85%). Bands A+B were combined because of low yields.2. Removal of adherent cells (5): Three milliliters of warmed cell suspension (1 to 3 X 107) were incubated on a glass wool column (5) at 370 for 30 min, then eluted with 30 ml of warm medium (nonadherent fraction). Slightly adherent cells were removed by gentle plunging. The wool was roughly shaken in a siliconized bottle with 50
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