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Biomedical subjects

D M Marcus

Publications and source records attributed to D M Marcus.

At least 127 records · Page 7Linked to original sources

Breast carcinoma-associated antigen. An immunocytochemical study.

An antiserum raised against a high molecular weight fraction of a medullary carcinoma of the breast was used to study breast carcinomas by means of an indirect unlabeled antibody peroxidase-antiperoxidase (PAP) technic. Strong cytoplasmic staining was demonstrated in all of 30 breast cancers tested and in one of three intraductal papillomas. Weak cytoplasmic staining was detected in four of nine fibroadenomas, two malignant melanomas, and one of six carcinomas of the colon. No cytoplasmic staining was seen in 24 other specimens of breast tissue, including 12 specimens from patients who had fibrocystic disease and five fibroadenomas, or in 17 extramammary malignant tumors. The antiserum appears to identify a breast carcinopma-associated antigen.

Antigens, Neoplasm↗

Preparation and properties of antibodies to GD3 and GM1 gangliosides.

Gangliosides GD3 and GM1 were coupled to proteins by their carboxyl groups and antisera were raised against the complexes. Anti-ganglioside antibodies were isolated by affinity chromatography on ganglioside-aminopropyl silica gel columns, and the specificity of the antibodies was determined by a quantitative microcomplement fixation assay. Antibodies to GD3 were highly specific and did not crossreact with GM3, lactosyl ceramide, or other glycolipids. Purified antibodies to GM1, in contrast, crossreacted with asialo-GM1, GD1b, and, to a lesser extent, GM2 and asialo-GM2. A derivative of GM1, containing a C-7 sialic acid residue produced by periodate oxidation, reacted with the ani-GM1 antibodies almost as readily as with GM1. The specificities of anti-GM1 antibodies elicited by the covalent ganglioside-protein complexes were similar to those produced by immunization with noncovalent complexes of GM1 and methylated bovine serum albumin. The ganglioside-protein complexes described here should be useful for preparing antibodies to polysialogangliosides that contain neuraminidase-sensitive linkages.

Antibodies↗

A new Pk phenotype in the P blood group system.

A healthy 22-year-old woman was noted to have erythrocytes of the Pk phenotype: a strong Pk antigen, no detectable P antigen and anti-P antibody in her serum. Her erythrocytes contained four to six times as much Pk glycolipid (globotriaosylceramide or CTH) and approximately half as much P glycolipid (globotetraosylceramide or globoside) as normal red cells. The structures of CTH and globoside were characterized by analysis of permethylated sugars and complement fixation, in addition to chromatographic mobility and sugar composition. Inasmuch as the erythrocytes of two Pk individuals that were analysed previously (Marcus et al., 1976) contained no detectable globoside, these abnormalities appear to represent a new phenotype in the P blood group system.

Adult↗

An improved technique for separation of neutral glycosphingolipids by high-performance liquid chromatography.

We have developed a high-performance liquid chromatographic (HPLC) procedure for separation of O-acetyl-N-p-nitrobenzoyl derivatives of six neutral glycosphingolipids: glucosylceramide, lactosylceramide, globotriaosylceramide, lactotriaosylceramide, globotetraosylceramide, and neolactotetraosylceramide. The recoveries of glucosylceramide and globotetraosylceramide for the derivatization procedure and HPLC analysis were approximately 75%, and one nanomole of glycolipid could be detected. The procedure was used for analysis of human erythrocyte neutral glycolipids.

Chromatography, High Pressure Liquid↗

Characterization of ferritin from human placenta. Implications for analysis of tissue specificity and microheterogeneity of ferritins.

Mammalian ferritins can be resolved into multiple components by isoelectric focusing, and each tissue contains a characteristic subset of isoferritins. Ferritin isolated from human liver was compared to acidic ferritin isolated from mid-gestational human placenta to define a structural basis for ferritin heterogeneity. Placenta ferritin contained several major bands with isoelectric points in the range of pI = 4.7-5.0 which were more acidic than the predominant isoferritins of human liver. Ferritin from each tissue was resistant to denaturation by 10 M urea and appeared to be identical by electron microscopy. Circular dichroism measurements revealed that placenta ferritin had substantially less ordered secondary structure than liver ferritin. Both types of ferritin contained only two subunits when analyzed by electrophoresis in sodium dodecyl sulfate gels, but isoelectric focusing of dissociated subunits in urea revealed 6-7 different components. In this system, placenta ferritin was enriched in the more acidic subunits and it completely lacked the most basic subunits noted in liver ferritin; placental ferritin had no unique components. Differences in isoelectric points among assembled ferritins from these two tissues appear to result from different proportions of these acidic and basic subunits.

Chemical Phenomena↗

Abnormal glycolipid composition of erythrocytes with a weak P antigen.

The erythrocytes of a normal man were agglutinated more weakly than normal cells by several anti-P1PPK sera, and exhibited a decreased capacity to absorb these antibodies. Analysis of his erythrocyte glycosphingolipids revealed that the globoside (P antigen) content was less than 25% of normal, and trihexosyl ceramide (the PK antigen) was 30-40% of normal. The ganglioside content of his erythrocytes was approximately four times normal and sialosylparagloboside was increased about sixfold. It appears that his erythrocytes was unable to synthesize normal quantities of trihexosyl ceramide, and that these serological and chemical features constitute a new phenotype in the P blood group system.

Adult↗

Glycolipid markers of murine lymphocyte subpopulations.

We have shown previously that purified antibodies to ganglioside GM1 react with peripheral T cells and most thymocytes in several strains of mice, independent of Thy-1 phenotype. GM1 and the Thy-1.2 antigen cap independently on C3H thymocytes, which provides additional evidence that GM1 is not the Thy-1.2 antigen. In C3H and nude mice antibodies to GM1 also react with a population of cells, comprising about 25% of lymphocytes from lymph nodes or spleen, that bear surface immunoglobulin. After removal of immunoglobulin from these cells by digestion with proteolytic enzyme, the GM1+ cells regenerate their surface immunoglobulin during 18 hr in culture, which indicates that these double-labeled cells synthesize their surface immunoglobulin. Protease treatment of lymphocytes reveals receptors for antibodies to GM1 on most cells. These data indicate that T and B cells differ in the accessibility of GM1 to antibody, and not necessarily in their content of GM1. Purified antibodies to asialo GM1 react with mature T cells in all strains of mice tested. In contrast to anti-GM1, these antibodies do not react with most thymocytes, with immunoglobulin-bearing lymphocytes of C3H or nude mice, nor with pronase-treated B cells.

Animals↗

Glycosphingolipids of purified human lymphocytes.

Biochemical analysis of the glycosphingolipids (GSLs) of human lymphocytes revealed qualitative and quantitative variations among purified lymphocytes from different tissues. The major neutral GSLs of tonsil lymphocytes are glucosyl ceramide (CMH), lactosyl ceramide (CDH), trihexosyl ceramide (CTH), and globoside. Thymocytes and peripheral blood lymphocytes (PBL) contain only traces of CTH and globoside, and PBL contain more CMH and CDH per cell than tonsil lymphocytes. Thymocytes and PBL contain relatively large amounts of more complex neutral GSLs that are present in only trace amounts in tonsil lymphocytes. Peripheral blood lymphocytes contained three and five times more lipid-bound sialic acid than thymocytes and toncil lymphocytes, respectively. Thymocytes and PBL contained mostly hematoside, whereas tonsil lymphocytes contained more complex gangliosides in addition to hematoside. The observed differences in GSL content among these cells may be related to their content of B cells, which comprise approximately 50% of tonsil lymphocytes, 10% of PBL and 0-2% of thymus cells, and/or the known differences in functional capacities of cells in different lymphoid organs. These findings suggest that cell surface GSLs may serve as markers for identification of functional subpopulations of human lymphocytes.

Carbohydrates↗

Identification of sialosylparagloboside as the erythrocyte receptor for an 'anti-p' antibody.

A human antibody which preferentially agglutinates p erythrocytes is inhibited specifically by the glycolipid sialosylparagloboside, NeuNAc (alpha,2 leads to 3)Gal(beta,1 leads t0 4)GlcNAc(beta,1 leads to 3)Gal(beta,1 leads to 4)Glc-Cer and forms a precipitin band with this compound in agarose gel. Erythrocytes treated with neuraminidase are no longer agglutinated by this antibody, but the agglutination is not affected by papain treatment. Sialosylparagloboside is not a biosynthetic precursor of any of the P antigens, but it contains the lacto-N-neotetraose structure which is also present in the P1 glycolipid.

Binding Sites, Antibody↗

The reactions of antibodies to paragloboside (lacto-N-neotetraosyl ceramide) with human erythrocytes and lymphocytes.

Paragloboside (lacto-N-neotetraosyl ceramide) is a biosynthetic precursor of the ABH and P1 blood group glycosphingolipids and of one class of gangliosides. This study concerns the specificity of rabbit anti-paragloboside antibodies, andtheir reactions with human erythrocytes and lymphocytes. Lacto-N-neotetraose was approximately 10 times as effective as lacto-N-tetraose in inhibiting complement fixation by purified IgG antibodies to paragloboside. The purified antibodies cross-reacted weakly with lactosyl ceramide, asialo GM1, and lacto-N-neotetraosyl polylysine. The purified IgG and IgM antibodies exhibited weak hemagglutinating activity which was enhanced by treatment of erythrocytes with papain or neuraminidase. In contrast to a macroglobulin described by Tsai et al. (J. Immunol. 117:717, 1976), which also binds paragloboside and lacto-N-neotetraose, anti-paragloboside antibodies reacted equally well with normal adult, O1, and umbilical cord erythrocytes. The IgG antibodies to paragloboside reacted with about 60% of B lymphocytes of peripheral blood and 5 to 10% of T cells. The relationship between antibodies to paragloboside and other lectins and antibodies directed against beta-galactosyl groups, including anti-T (Thomsen-Friedenreich) antibodies, is discussed.

Agglutinins↗

Binding of N-acetylgalactosamine-containing compounds by a human IgM paraprotein.

A serum that contains a monoclonal human IgM paraprotein (McG) agglutinates protease-treated human erythrocytes and binds glycosphingolipids that possess a terminal nonreducing N-acetylgalactosaminyl residue in either the alpha or beta anomeric configuration. The approximately equal reactivity of McG with both anomers of N-acetylgalactosamine was unexpected because most immunoglobulins that bind saccharides exhibit a marked preference for one anomeric configuration. The major receptor for this protein in normal group O erythrocytes is globoside, the blood group P antigen: Ga1NAc( beta, 1 leads to 3)Gal(alpha,1 leads to 4)Gal(b,1 lead to 4)G1c-ceramide. Erythrocytes of the rare Pk and p phenotypes lack globoside, and the former are not agglutinated by McG. The McG receptor in p erythrocytes, which are agglutinated as strongly as normal cells, appears to be a crossreactive glycolipid that contains a much larger number of sugar residues than globoside.

ABO Blood-Group System↗

Inhibition of leukocyte migration by human breast-cancer-associated antigens.

The existence of CMI to tumor-associated antigens present in 3 M KCl extracts of breast carcinomas was demonstrated in a group of breast cancer patients by the leukocyte migration inhibition (LMI) assay. When crude KCl extracts were tested, 3 of 5 breast cancer patients gave a positive response to autologous tumor extracts. Eleven of 20 gave a positive response to allogeneic extracts as compared to 3 of 22 controls (including 6 patients with benign breast disease, 7 with non-mammary cancers and 9 normal controls). Extracts of fibrocystic disease tissue gave positive LMI tests in 2 of 5 breast cancer patients, suggesting the presence of antigenic cross-reactivity between benign and malignant breast disease. An extract of a medullary carcinoma of breast was fractionated on Sephadex G-200 and the effluent pooled into three fractions. The high molecular weight fraction produced LMI in 11 of 22 breast cancer patients and in 1 of 19 controls, including patients with benign breast disease, other cancers and normal individuals. The low molecular weight fraction produced LMI in both the benign (4 of 6) and the malignant breast disease (6 of 20) patients, but not in the controls (0 of 12). A simple fractionation technique has thus separated "cancer-specific" from "organ-specific" activity. Sephadex G-200 fractions were active at a much lower protein concentration than the crude 3 M KCl extracts.

Adenofibroma↗