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Biomedical subjects

D M Marcus

Publications and source records attributed to D M Marcus.

At least 91 records · Page 5Linked to original sources
PubMed1989

Measurement and significance of antibodies against GM1 ganglioside. Report of a workshop, 18 April 1989, Chicago, IL, U.S.A.

Twelve laboratories from the United States, Canada, France, Italy and Switzerland participated in a workshop to compare assays used to measure anti-GM1 antibodies, and to discuss the clinical significance of these antibodies. A panel of test samples containing varying amounts of anti-GM1 antibody was prepared by mixing varied proportions of normal serum with a serum containing a monoclonal IgM antibody that bound GM1 ganglioside. Enzyme-linked immunosorbent assay (ELISA) data were supplied by eight laboratories and ten laboratories classified the sera as negative, weakly or strongly positive. Most laboratories correctly identified the two samples that contained the highest quantities of antibody, but there was considerable disagreement on the classification of the three samples with moderate or small amounts of antibody. The sensitivity of the assays varied considerably. The more sensitive assays did not use detergent in the washing buffers, and incubated the human serum with the antigen at 4 degrees C overnight. Several investigators have identified a subset of patients with lower motor neuron disease or multifocal neuropathy who have high titers of anti-GM1 antibodies. Many patients with neurological and non-neurological diseases have low to moderate levels of anti-GM1 antibodies, and the significance of these antibodies is unclear. There was general agreement that standardization of the ELISA assays is urgently required, and that distribution of a reference high-titered antiserum would facilitate this process.

Antibodies↗
PubMed1988

Structure of a ganglioside with Cad blood group antigen activity.

The Cad antigen is a rare erythrocyte blood group antigen expressed on both sialoglycoprotein and ganglioside structures. It is related both serologically and biochemically to the Sda blood group antigen expressed on over 90% of Caucasian erythrocytes. We reported previously that Cad erythrocytes contain a novel ganglioside that binds Helix pomatia lectin and inhibits human anti-Sda antibody. We have now purified the Cad ganglioside and determined its structure. The ganglioside contained Glc-Gal-GlcNAc-GalNAc-NeuAc in a molar ratio of 1.00:1.94:0.95:0.93:1.05. Its chromatographic mobility was between that of GM1 and GD3. After treatment with beta-hexosaminidase (human placenta Hex A), the product migrated with 2-3-sialosylparagloboside (IV3NeuAcnLc4OseCer), it no longer bound H. pomatia lectin, and it acquired the ability to bind an antibody to sialosylparagloboside. Treatment of this material with neuraminidase (Vibrio cholerae) yielded a product with the mobility of paragloboside (nLc4OseCer) that bound monoclonal antibody 1B2, which is specific for terminal N-acetyllactosaminyl structures. Treatment of the Cad ganglioside with Arthrobacter ureafaciens neuraminidase yielded a product reactive with monoclonal antibody 2D4, which is specific for terminal GalNAc beta (1-4)Gal structures. These data provide strong evidence that the Cad ganglioside structure is GalNAc beta (1-4)[NeuAc alpha (2-3)]Gal beta (1-3)Gal beta (1-4)GlcCer. 1H NMR analysis also supports the conclusion that the terminal GalNAc is linked beta (1-4) to Gal. High-performance thin-layer chromatographic ganglioside patterns from three blood group Cad individuals showed a direct correlation between the quantity of Cad ganglioside and the strength of Cad antigen expression on the erythrocytes, as measured by hemagglutination.(ABSTRACT TRUNCATED AT 250 WORDS)

Blood Group Antigens↗
PubMed1988

Monoclonal antibodies that bind to galactosylgloboside (SSEA-3 antigen).

The 3-fucosyllactosamine (3-FL) antigenic determinant, Gal(beta 1-4)[Fuc(alpha 1-3)]-GlcNAc-R, is very immunogenic in BALB/c mice. A panel of monoclonal antibodies was raised against a novel glycosphingolipid that contains a terminal 3-FL structure (structure 1): (formula: see text) Surprisingly, most of these antibodies reacted with the internal galactosylgloboside structure rather than with the terminal 3-FL epitope. The specificities of two of these antibodies, 5A3 and 8A7, were analyzed by enzyme-linked immunosorbent assay and immunostaining procedures, and compared with MC631 (anti-SSEA-3), the only other monoclonal antibody known to bind to galactosylgloboside. 5A3 and 8A7 bound equally well to structure 1 and to galactosylgloboside. These antibodies also bound to structure 1 from which the terminal galactose, or the galactose and fucose, had been removed, but not to globoside or sialylated galactosylgloboside. In contrast, MC631 did not bind to structure 1 but, as described previously, did bind to globoside and sialylated galactosylgloboside. Galactosylgloboside is a developmentally regulated antigen of mouse embryos, human teratocarcinomas, and mouse embryonic brain. Antibodies 5A3 and 8A7 will complement MC631 in the analysis of the distribution and regulation of galactosylgloboside.

Antibodies, Monoclonal↗
PubMed1988

Sequences of the VH and VL regions of murine monoclonal antibodies against 3-fucosyllactosamine.

Many mAb that bind the carbohydrate antigenic determinant 3-fucosyl-lactosamine (3-FL), Gal beta 1-4[Fuc alpha-3]GlcNAc-R have been raised in BALB/c mice, and we are studying the structure and regulation of these antibodies. In this report, we present the first information about their amino acid sequences and the Ig gene segments used to encode them. V regions of the H and L chains of three anti-3-FL antibodies, PMN6, PMN29, and PM81, were sequenced by a combination of mRNA and amino acid sequencing. The L chain sequences of PMN6 and PM81 antibodies indicate that their VK and JK regions are encoded by VK24B and JK1 germ-line genes, respectively. The nucleotide and amino acid sequences of the H chains suggest that the three anti-3-FL antibodies are encoded by the VH441 gene segment of the X24 VH family, and this conclusion was supported by Southern filter hybridization with VH441 and JH3-JH4 probes. PMN29 has at least 11 amino acid substitutions, which is an unusually large amount of somatic mutation for an IgM antibody. Previous analyses of BALB/c genomic libraries with VHX24 and VH441 probes make it unlikely that this VH family contains additional germ-line genes, but this possibility cannot be excluded. All three antibodies use the DQ52 and JH4 gene segments. The single VH and VL gene segments used to encode the anti-3-FL antibodies is in contrast to the multiple VH and VL segments used by antibodies against other carbohydrate Ag such as alpha 1-6 dextran and group A streptococcal carbohydrate. VH441 also encodes the VH regions of antibodies against galactan and levan (beta 2-6 fructosan). The similarities among VH segments of antibodies against 3-FL, levan, and galactan, and the striking differences in their CDR3 sequences, suggest that CDR3 plays an important role in the formation of the Ag binding site. The use of a single VH segment from the smallest VH gene family by antibodies against at least three different carbohydrate determinants is noteworthy. It raises the possibility that the amino acid sequence encoded by VH441 has some general structural features that make it particularly well adapted for binding to carbohydrate sequences.

Amino Acid Sequence↗
PubMed1988

Zoster-related bilateral acute retinal necrosis syndrome as presenting sign in AIDS.

The acute retinal necrosis (ARN) syndrome has recently been associated with intraocular infections with one or more members of the herpesvirus family. There have been 14 cases in the literature linking ARN with a preceding or subsequent herpetic dermatitis. We report the development of bilateral ARN (BARN) after unilateral Herpes zoster ophthalmicus as the presenting sign of acquired immunodeficiency syndrome (AIDS) in a previously healthy man. The development of BARN after diffuse Herpes simplex dermatitis in AIDS patients is also discussed. These cases further illustrate the central role of the herpes-virus family in the etiology of ARN and alert the clinician to a new presenting sign for AIDS.

Acquired Immunodeficiency Syndrome↗
PubMed1987

Glycosphingolipids of human umbilical vein endothelial cells and smooth muscle cells.

Glycosphingolipids (GSLs) represent an important class of immunogens and receptors. Although cell surface antigens and receptors of endothelial cells (ECs) have been the subject of extensive biochemical investigation, no information is available about their GSLs. We report here the characterization by chromatographic and immunological techniques of GSLs of cultured human umbilical vein ECs and, for comparison, umbilical vein smooth muscle cells (SMCs). The most abundant neutral GSLs of both cell types were lactosylceramide, Gb3, and Gb4, and both cells contained complex lacto and globo series compounds. Immunostaining revealed that ECs, but not SMCs, contained long chain GSLs bearing a type 2 blood group H determinant. ECs also contained more long chain GSLs bearing an unsubstituted terminal lactosamine structure than SMCs. Labeling with galactose oxidase/NaB3H4 demonstrated that neutral glycolipids that contained three or more sugars were accessible on the cell surface. The major gangliosides of both cell types were GM3 and IV3NeuAcnLc4. Immunostaining following neuraminidase treatment revealed that most of the long chain gangliosides in both types of cells contained a lacto core structure, and that ganglio series compounds were more abundant in SMCs than ECs. Gangliosides that contain a polyfucosyllactosamine core and a globo core were also present in both cell types. These results demonstrate that endothelial and smooth muscle cells contain a large diversity of GSL structures, and provide the basis for investigation of the role of these GSLs as cell surface antigens and receptors for blood components.

Antibodies↗
PubMed1987

Studies of the mechanism by which gangliosides inhibit the proliferative response of murine splenocytes to concanavalin A.

Gangliosides are known to inhibit the proliferative response of murine and human lymphocytes to antigens and mitogens in vitro. In this study the response of murine spleen cells to concanavalin A (Con A) was used as a model system. Analysis of the cellular events by flow cytometry revealed that during the first 24 hr of culture the effect of gangliosides on Con A-treated cells was minimal. At 48 hr, however, more of the ganglioside-treated cells were in G0/G1, the cells contained more RNA, and fewer cells were in S phase. These data indicate that gangliosides inhibit the transition of the cells from G0/G1 into the S phase of the cell cycle. Expression of the interleukin 2 (IL-2) receptor, as measured by the binding of a monoclonal antibody to the receptor, was not inhibited by the gangliosides. Binding of 125I-labeled recombinant IL-2 to cells cultured for 48 hr with Con A was inhibited by ganglioside GD1a but not by asialo GM1. Inhibition was much more effective if the gangliosides were preincubated with IL-2 before addition of cells, but no inhibition was observed if the cells were preincubated with gangliosides and the unbound gangliosides were washed out prior to addition of the IL-2. These data suggest that interference with the binding of IL-2 to the high-affinity IL-2 receptor of activated T lymphocytes plays an important role in the inhibition of Con A-induced proliferation.

Animals↗
PubMed1987

New abnormalities in the morphology, cell surface receptors, and electrolyte metabolism of In(Lu) erythrocytes.

The In(Lu) phenotype is inherited as an autosomal dominant trait and is characterized by suppression of the Lutheran, P1, i, and Aua erythrocyte blood group antigens. We have developed a monoclonal antibody (L21) that strongly agglutinates all erythrocytes except In(Lu), and we have identified eight In(Lu) individuals among 42,000 blood donors tested. Studies of two families confirmed the dominant mode of inheritance and revealed several new features of this phenotype. The erythrocytes of all five affected individuals from the two families exhibited diminished hemagglutination by the lectin concanavalin A, although they reacted normally with several other lectins. The erythrocytes of two affected individuals in one family exhibited marked acanthocytosis. The erythrocytes of the proposita of the other family exhibited a mild degree of poikilocytosis, but the cells of the other two affected individuals in this family had normal morphology. The osmotic fragility of fresh In(Lu) erythrocytes was normal, but after incubation for 24 hours at 37 degrees C in plasma the In(Lu) cells exhibited a marked increase in resistance to osmotic lysis. During the incubation period the erythrocytes lost K+ and their total cation content was diminished. These data indicate that in addition to the suppression of blood group antigens noted previously, the In(Lu) phenotype includes a variety of morphological abnormalities and a defect in electrolyte metabolism. The use of L21 and similar monoclonal antibodies provides a more sensitive means of detecting In(Lu) erythrocytes than typing with human anti-Lub antisera.

Antibodies, Monoclonal↗
PubMed1986

The occurrence of anti-3-fucosyllactosamine antibodies and their cross-reactive idiotopes in preimmune and immune mouse sera.

The carbohydrate determinant 3-fucosyllactosamine (3-FL), Gal(beta 1-4)[Fuc alpha 1-3]GlcNAc-R, which is also known as SSEA-1 or as the X determinant, is very immunogenic in BALB/c mice. Many monoclonal antibodies directed against this structure have been obtained by immunization of mice with murine teratocarcinomas and human carcinomas and myeloid cells. We have undertaken an analysis of the regulation of these antibodies and of their idiotypic, structural, and genetic diversity. We have described previously the preparation of syngeneic monoclonal anti-idiotypic antibodies (6C4 and 6B1) that reacted with 50% of a panel of monoclonal anti-3-FL antibodies. In this study, we have examined the occurrence of anti-3-FL antibodies and their cross-reactive idiotopes in the sera of unimmunized and immunized mice. All BALB/c sera examined had more naturally occurring antibodies against 3-FL than against four other glycolipid antigens, and the 6C4 and 6B1 idiotopes were also detected in these sera. Approximately 25% of the anti-3-FL antibodies could be removed from a pool of BALB/c sera by a 6C4 affinity column, and the eluate from this column exhibited strong binding to 3-FL antigens. After a single i.v. injection of a 3-FL-positive glycolipid coated on Salmonella minnesota, anti-3-FL titers rose in BALB/c mice. The level of 6B1 idiotope rose in most mice, but the idiotope level showed no correlation with anti-3-FL titers. Naturally occurring antibodies against 3-FL were also noted in the sera of AKR, C3H/He, DBA/2, BALB/c-nu/nu, and CBA/CaHN mice but not in C57BL/6, SM, or CBA/N mice. A single i.v. injection of antigen elicited an antibody response in C3H/He mice but not in C57BL/6, SM, or DBA/2 mice. These data indicate that several strains of mice have more naturally occurring IgM antibodies against the 3-FL structure than against other glycolipids, and that this response may be genetically regulated. The 6C4 and 6B1 cross-reactive idiotopes that we have identified previously on monoclonal antibodies are also present in preimmune and immune sera. The existence of a population of B lymphocytes that are primed against the 3-FL determinant accounts in part for the immunogenicity of this structure.

Animals↗
PubMed1986

Idiotypic determinants of monoclonal antibodies that bind to 3-fucosyllactosamine.

Many monoclonal antibodies that react with the lacto-N-fucopentaose III (LNF III) antigenic determinant, Gal beta 1-4(Fuc alpha 1-3)GlcNAc beta 1-3Gal beta 1-4Glc, have been described recently. The terminal trisaccharide of this determinant, fucosyllactosamine, is present on glycolipids and glycoproteins and on the surface of granulocytes, monocytes, and other cells. To study the structural and genetic diversity of these antibodies, syngeneic anti-idiotypic monoclonal antibodies were produced in BALB/c mice against PMN 6, a monoclonal antibody directed against this sequence. Anti-idiotypic antibodies 6B1 and 6C4 reacted with 50% of a panel of 20 anti-LNF III monoclonal antibodies, whereas 6A3 reacted strongly only with PMN 6. This indicates that the determinants recognized by 6C4 and 6B1 represent major cross-reactive idiotopes of this family of antibodies. The binding of idiotypic antibodies to a glycolipid bearing this antigenic determinant was completely inhibited by the three anti-idiotypic antibodies, 6A3, 6B1, and 6C4. The idiotopes could be demonstrated on the heavy chain of the monoclonal antibodies by an antibody transfer technique when mild reducing conditions were employed, but a high concentration of reducing agent destroyed the idiotypic determinants. This suggests that the anti-idiotypic antibodies recognize conformational structures expressed on the heavy chain molecules. The binding of 18 monoclonal antibodies to two glycolipid antigens and to a fucosyllactosamine-bovine serum albumin conjugate was compared. Antibodies that possessed the 6C4 cross-reactive idiotope bound to fucosyllactosamine-bovine serum albumin more weakly than idiotype-negative antibodies (p = 0.001). This suggests that the 6C4-positive antibodies might represent germline structures.

Animals↗
PubMed1985

Spontaneous and plasma factor-mediated transfer of pyrenyl cerebrosides between model and native lipoproteins.

A series of pyrenyl glucocerebrosides was synthesized by reacylation of psychosine with pyrene-labeled fatty acids having 3-11 methylene units. When incorporated into model high-density lipoproteins consisting of dimyristoylphosphatidylcholine-apolipoprotein A-II complexes and incubated with unlabeled complexes, these lipids exhibited spontaneous transfer. Half times of transfer varied from 1.5 min to 365 min at 37 degrees C. The logarithm of the rate of transfer was linearly related to the number of fatty acyl methylene units and HPLC retention time. Transfer occurred by passage of lipid monomers through the aqueous phase. Spontaneous transfer of the glycolipids also occurred when they were incorporated into native high-density lipoproteins. Rates of transfer between native high-density lipoprotein particles were higher than those observed between model high-density lipoprotein particles. A partially purified lipid exchange protein from plasma, as well as unfractionated lipoprotein-deficient serum, stimulated the transfer of fluorescent glycolipid between model high-density lipoprotein or native high-density lipoprotein and low-density lipoprotein 2-24 fold. The protein also stimulated the transfer of tritiated ganglioside GM3 between native low-density lipoprotein and high-density lipoprotein. This protein may play a role in glycolipid exchange in vivo.

Apolipoprotein A-II↗
PubMed1985

Glycosphingolipids of human plasma.

A number of glycosphingolipids, including 10 gangliosides, not previously identified in human plasma have been characterized. The plasma contains 2 micrograms of lipid-bound sialic acid/ml plasma and 54% of the gangliosides are monosialo, 30% disialo, 10% trisialo, and 6% tetrasialo. Individual glycosphingolipids were purified by high-performance liquid chromatography and thin-layer chromatography, and were characterized on the basis of their chromatographic mobility, carbohydrate composition, hydrolysis by glycosidases, methylation analysis, and immunostaining with anti-glycosphingolipid antibodies. The monosialogangliosides were identified as GM3, GM2, sialosyl(2-3)- and sialosyl(2-6)lactoneotetraosylceramides, sialosyllacto-N-nor-hexaosylceramide, and sialosyllacto-N-isooctaosylceramide. The major gangliosides in the polysialo fractions contained a ganglio-N-tetraose backbone and were identified as GD3, GD1a, GD1b, and GQ1b. The most abundant neutral glycosphingolipids were glucosyl, lactosyl, globotriaosyl, globotetraosyl and lactoneotetraosylceramides. The other neutral glycosphingolipids, tentatively identified by immunostaining with monoclonal antibodies, contained H1, Lea, Leb, and lacto-N-fucopentose III (X hapten) structures.

Carbohydrate Sequence↗
PubMed1984

Cytotoxic anti-T cell antibodies in children with juvenile rheumatoid arthritis.

The object of this investigation was to determine the prevalence of anti-T cell antibodies in 66 children with various connective tissue diseases. Anti-T cell antibodies were found in 43/44 juvenile rheumatoid arthritis (JRA) patients (mean cytotoxicity 15.0%) and in 10/10 children with systemic lupus erythematosus (mean cytotoxicity 20.0%), but in only 1/15 normal controls and in none of 12 children with other arthritides. There was no significant difference in mean percent cytotoxicity among the JRA subclasses. In the JRA patients, the percent cytotoxicity was positively correlated with the erythrocyte sedimentation rate (P = 0.01), but not with the presence or absence of rheumatoid factor, antinuclear antibodies, or immune complexes. The sera of 3 JRA patients repeatedly inhibited the stimulation of normal lymphocytes by mitogens and antigens by 47-99% (measured by the incorporation of 3H-thymidine into DNA) when added to the culture system in the first 24 hours; normal sera did not. Sera from patients with JRA have increased reactivity with mitogen-activated lymphocytes and T cells compared with unstimulated cells as determined by flow cytometry. The expression of the "JRA antigen" requires protein synthesis but not DNA synthesis or cell division. We conclude that the majority of patients with active JRA have cytotoxic anti-T cell antibodies and that in selected patients, these antibodies may play a role in regulation of the immune response.

Antibodies↗