Recombinant vaccinia virus vaccine against the human melanoma antigen p97 for use in immunotherapy.

We have constructed a recombinant vaccinia virus, v-p97NY, which expresses the human melanoma-associated glycoprotein p97. Immunization with v-p97NY could induce humoral and cell-mediated immunity to p97, including delayed-type hypersensitivity, in mice and in two of two monkeys (Macaca fascicularis). The fact that an immune response was induced also in monkeys is important because normal cells from monkeys, but not from mice, express a low level of cross-reactive p97.

Antibody-mediated killing of human tumor cells by attached effector cells.

Cultured human melanoma, lung carcinoma, and colon carcinoma cells were isotope labeled and incubated with a combination of effector cells and mouse monoclonal antibodies to tumor-associated cell surface antigens. The former were derived from the peritoneal cavity of mice or from peripheral blood of healthy human subjects. Monoclonal antibodies MG-21, 96.

Production of a mouse-human chimeric monoclonal antibody to CD20 with potent Fc-dependent biologic activity.

Mouse monoclonal antibody 2H7 recognizes the CD20 cell surface phosphoprotein that is expressed in normal as well as malignant B cells. CD20 may be a useful target for therapy of B cell lymphomas, since damaged normal B cells can be replaced by their antigen-negative precursors. Monoclonal antibody 2H7 is an IgG2b (kappa) immunoglobulin which cannot mediate antibody-dependent cellular cytotoxicity with human lymphocytes or complement-dependent cytotoxicity with human serum.

Immunotherapy of murine sarcomas with auto-anti-idiotypic monoclonal antibodies which bind to tumor-specific T cells.

Hybridomas producing monoclonal antibodies (mAb) were obtained from BALB/c mice immunized against either of two transplanted, chemically induced syngeneic sarcomas, MCA-1490 or MCA-1511. Two mAb, 4.72 and 5.

Induction of calcium flux and enhancement of cytolytic activity in natural killer cells by cross-linking of the sheep erythrocyte binding protein (CD2) and the Fc-receptor (CD16).

Binding of the anti-cluster of differentiation (CD) 2 monoclonal antibody 9-1 causes an increase in the concentration of cytoplasmic-free calcium ([Ca2+]i) in cultured CD3-/CD16+ natural killer (NK) cells. This response did not occur in cultured CD3+/CD16- cytotoxic T lymphocytes (CTL). Anti-CD16 antibodies could partially block the calcium response when NK cells were stimulated with intact antibody 9-1, and antigen-binding fragment F(ab')2 of antibody 9-1 did not produce a calcium response.

Cholesterol biosynthesis in nonketotic diabetics before and during insulin therapy.

A previous study demonstrated that the net steroid balance, i.e., the total rate of cholesterogenesis, was within the normal range in insulin-treated patients with both insulin-dependent and noninsulin-dependent diabetes mellitus (NIDD).

Increased delivery of tumor-specific monoclonal antibodies to brain after osmotic blood-brain barrier modification in patients with melanoma metastatic to the central nervous system.

We evaluated the delivery of melanoma-specific radiolabeled monoclonal antibody (MAb) Fab fragments in a pilot study of three patients with melanoma metastatic to the central nervous system. Tumor samples demonstrated excellent immunohistochemical reactivity with Fab 96.5, specific for a 97,000-molecular-weight melanoma antigen (p97), or Fab 48.

Chimeric mouse-human IgG1 antibody that can mediate lysis of cancer cells.

A chimeric mouse-human antibody has been created that recognizes an antigen found on the surface of cells from many carcinomas. Immunoglobulin constant (C) domains of the mouse monoclonal antibody L6, C gamma 2a and C kappa, were substituted by the human C gamma 1 and C kappa by recombining cDNA modules encoding variable or C domains. The cDNA constructs were transfected into lymphoid cells for antibody production.

Effects of a high-protein and low-fat diet vs a low-protein and high-fat diet on blood glucose, serum lipoproteins, and cholesterol metabolism in noninsulin-dependent diabetics.

Six middle-aged patients with noninsulin-dependent diabetes and six normoglycemic control subjects were fed protein-rich and fat-poor (diet A) or protein-poor and fat-rich food (diet B). The patients were hyperglycemic, VLDL triglycerides levels were higher, and HDL cholesterol levels lower than corresponding findings in control subjects. Bile acid formation and biliary lipid composition did not differ between the two groups, but net steroid balance in the patients was elevated by a factor of approximately 2.

Preliminary studies of monoclonal antibody lymphoscintigraphy in malignant melanoma.

Lymphoscintigraphy was performed at 3 and 20 hr following subcutaneous injection of 131I anti-melanoma antibody (Fab) in 11 patients who had surgical resection of lymph nodes (neck, axilla, groin) at 24 hr for suspected metastatic melanoma. Comparable amounts of 125I nonspecific control antibody (Fab) were co-administered. Six patients had nodal metastases and three showed positive images at both time periods.

Tumor-specific idiotopes on suppressor factors and suppressor cells revealed by monoclonal anti-idiotope antibodies.

Two monoclonal anti-idiotope antibodies, previously found to induce tumor-specific cell-mediated immunity in mice, were examined for their relationship to tumor-associated suppressor factors (SF), produced in culture by spleen cells from tumor-bearing mice or present in sera from such mice. A leukocyte adherence inhibition assay was used to detect cellular immunoreactivity to tumor antigens and its inhibition by SF, using peritoneal cells from mice bearing tumor or sensitized with anti-idiotope antibody. The SF were specifically absorbed by the corresponding anti-idiotope antibodies coupled to a solid phase and were recovered by elution.

Anti-idiotypic antibodies and the induction of specific tumor immunity.

Immunization with anti-idiotypic antibodies is a strategy which, with variable success, can be used to elicit or amplify antigen-specific immune response. This article discusses the manipulation of specific idiotypes in anti-tumor immunity, emphasizing the appropriate consideration of genetic restriction, the choice of idiotype specificity, and the route of immunization. Two independent pathways are outlined: One uses anti-idiotypic antibodies to select and amplify tumor-specific T and B cells via their preexisting antigen-specific receptors, and the other uses anti-idiotypes as primary internal image immunogens to elicit immune recognition of determinants shared by the anti-idiotype and by tumor-associated antigens.

The melanoma proteoglycan: restricted expression on microspikes, a specific microdomain of the cell surface.

A cell surface chondroitin sulfate proteoglycan associated with human melanomas and defined by mAb's F24.47 and 48.7 has been characterized biochemically and localized by indirect immunogold electron microscopy.

Cholesterol and bile acid metabolism in middle-aged diabetics.

Serum lipoproteins, bile and kinetics and net steroid balance were studied in 22 diet-treated and 5 insulin-treated patients with noninsulin-dependent diabetes mellitus, in 6 patients with insulin-dependent diabetes mellitus and in 15 normoglycemic controls. All subjects were middle-aged and the patients were hyperglycemic. Some of the diet-treated patients suffered from obesity and/or dyslipoproteinaemia mainly characterized by elevated levels of triglycerides and cholesterol in VLDL (very low-density lipoproteins).

Antitumor effects of L6, an IgG2a antibody that reacts with most human carcinomas.

Mouse monoclonal antibody L6 (IgG2a subtype) recognizes a ganglioside antigen expressed at the surface of cells from human non-small-cell lung carcinomas, breast carcinomas, and colon carcinomas. We now show that this antibody can lyse L6 antigen-positive human tumor cells in the presence of Leu-11b-positive human lymphocytes (i.e.

Monoclonal mouse antibodies raised against human lung carcinoma.

We have evaluated approximately 10,000 monoclonal antibodies (MoAb) resulting from 25 hybridizations of spleen cells from mice immunized with cells from human non-small cell lung carcinoma or fetal lung. The spleen cells were hybridized with NS-1 myeloma cells, and the resulting hybridomas were screened for production of MoAb to non-small cell lung carcinoma by binding assays with either cell extracts or cells growing in culture, followed by immunohistology on frozen sections. Fourteen MoAb had relative specificity for non-small cell lung carcinoma versus normal tissues.

Immunoreactivity assay for labeled anti-melanoma monoclonal antibodies.

A convenient, rapid, and reproducible assay was developed to evaluate the immunoreactivity of radiolabeled monoclonal antibodies against three different human melanoma-associated antigens, p97, a proteoglycan and a GD3 ganglioside. A cloned melanoma cell line (M 2669 CL 13) was selected as the target and, when fixed with paraformaldehyde, showed binding as good as or better than that obtained with live cells for the three antigens. Fixed cells retained good binding properties stored at 4 degrees C for over 6 mo.

Identification of a novel serum protein secreted by lung carcinoma cells.

The murine anti-human lung tumor monoclonal antibody L3 recognizes antigens found both in the medium of cultured carcinoma cells and in normal human serum. Sequential immunoprecipitation experiments indicate that the L3 antigen is also recognized by a previously described monoclonal antibody directed against a melanoma-associated antigen [Natali, P. G.

Primary structure of the human melanoma-associated antigen p97 (melanotransferrin) deduced from the mRNA sequence.

p97 is a cell-surface glycoprotein that is present in most human melanomas but only in trace amounts in normal adult tissues. To determine the structure of this tumor-associated antigen and to identify its functional domains, we have purified and cloned p97 mRNA and determined its nucleotide sequence. The mRNA encodes a 738-residue precursor, which contains the previously determined N-terminal amino acid sequence of p97.

Autoradiolysis of iodinated monoclonal antibody preparations.

We investigated the effects of ionizing radiation on the immunointegrity of antibody fragments (Fab) because large amounts of high specific activity 131I may damage the proteins. We found that 1000 Gy of external 137Cs gamma radiation was sufficient to destroy 80-90% of the immunointegrity of the initial preparation. This effect was also produced by internally added [131I]NaI in a quantity sufficient to provide the same radiation absorbed dose.