HLA-DR inhibits granulocytic differentiation without inducing apoptosis of CD34 cells.

Hematopoietic progenitors express HLA-DR molecules. However the significance of HLA-class II molecules on CD34+ cells remains unknown. The primary role of HLA-class-II molecules is antigen presentation although a second role, that of signal transduction, has been established in B cells. The role of HLA-DR in hematopoiesis was examined by determining the ability of CD34+ progenitor cells to differentiate to "Colony Forming Unit Granulocyte-Macrophage" (CFU-GM) and "Burst Forming Unit Erythrocyte" (BFU-E) in the presence of anti-HLA-DR monoclonal antibody. We observed a reduction in the number of CFU-GM which was due in part to down regulation of granulocyte rather than monocyte differentiation. These observations suggest that HLA-DR signals can regulate myelopoiesis. We point out especially the role of the HLA-DR molecule in the switch of CFU-GM between granulocyte or monocyte lineages. Although HLA-DR mediated apoptosis has been described in mature B lymphocytes apoptosis of CD34+ cells was excluded as a mechanism.

Lewis X structure increases cell substratum adhesion in L cells.

cDNAs of alpha-1,3-fucosyltransferase as well as alpha-1,3/4-fucosyltransferase were placed under the control of a beta-actin promoter and cytomegalovirus enhancer and were introduced into L cells. The transfected cells expressing Le(x) antigen showed increased cell substratum adhesion as compared to the antigen-negative cells, when they were cultured for 2 to 4 h in Dulbecco-modified minimum essential medium containing 0.05% bovine serum albumin. The increased cell substratum adhesion was completely inhibited by cycloheximide and anti-integrin antiserum, and partly by an RGD peptide and EGTA. These findings indicate that Le(x) structure promotes cell adhesion to substratum-bound material secreted by cells, and that the increased adhesion is mediated by integrin. Western blotting experiments have revealed an 85 kDa protein and a 50-60 kDa protein as carriers of Le(x) antigen in transfected cells. The latter is likely to be basigin, which is a member of the immunoglobulin superfamily and is considered to be an integrin-associated protein. We hypothesize that fucosylation of basigin enhances integrin-mediated cell substratum adhesion.

1H-and 13C-n.m.r. spectroscopy of 2-oxo-3-deoxy-D-glycero-D- galactononulosonic acid-containing oligosaccharide-alditols bearing Lewis X, Lewis Y and A-Lewis Y determinants isolated from the jelly coat of Pleurodeles waltl eggs.

Three acidic oligosaccharide-alditols carrying Lewis X, Lewis Y and A-Lewis Y determinants were isolated from the jelly coat of Pleurodeles waltl eggs. These compounds possess the following structures. Gal beta 1-4(Fuc alpha 1-3)GlcNAc beta 1-3[2-oxo-3-deoxy-D- glycero-D-galactononulosonic acid (KDN)alpha 2-6] GalNAc-ol; Fuc alpha 1-2Gal beta 1-4(Fuc alpha 1-3)GlcNAc beta 1-3(KDN alpha 2-6) GalNAc-ol and Fuc alpha 1-2(GalNAc alpha 1-3)Gal beta 1-4(Fuc alpha 1-3)GlcNAc beta 1-3(KDN alpha 2-6)GalNAc-ol. The complete 1H-n.m.r.-spectrum assignment for the three compounds and the 13C-n.m.r. analysis of the A-Lewis Y determinant-containing heptasaccharide are reported.

Sialyl-dimeric Lewis-X antigen expressed on mucin-like glycoproteins in colorectal cancer metastases.

Colorectal primary carcinomas and metastases from 20 Dukes' stage C or D patients were examined for the immunohistochemical localization and contents of various fucosylated N-acetyl-lactosamine oligomers by specific monoclonal antibodies (MAbs). MAbs used were SH1, specific for Lewis X antigen; FH4, specific for dimeric Lewis X antigen; FH6, specific for sialyl-dimeric Lewis X antigen; and KH1, specific for Lewis Y-Lewis X antigen. The distribution of the carbohydrate antigens identified by these MAbs was heterogeneous within the primary tumor as well as within the metastatic lesion. Examinations of serial sections indicated that areas within an individual tumor which were stained with one MAb were not always reactive with the other MAbs, although these four MAbs identify closely related structures. The degree of MAb reactivity with carcinoma sections was classified by percentage positive carcinoma cells, and primary tumors and metastases from the same patients were compared. An equivalent or higher proportion of carcinoma cells in the metastatic lesions were reactive with MAb FH6 than in the primary colon carcinomas, but each correlation was not seen with the other MAbs. Electrophoretic separation of tumor tissue extracts followed by staining with these MAbs revealed that a component having an approximate molecular weight of 1,000,000 is the major site for the binding of MAbs, FH6, FH4, and KH1. The electrophoretic mobility of the antigenic molecule on polyacrylamide gels as shown by direct MAb bindings was slightly different from that of a major sialomucin revealed by wheat germ agglutinin in the same tissues. MAb FH6 binding to a high molecular weight component was eliminated by prior treatment of the glycoprotein with mild acid or sialidase to remove sialic acid. Simultaneously, binding of MAb SH2, specific for dimeric Lex antigen, to this component increased. An extract was prepared from a liver metastasis, and high molecular weight components were isolated by gel filtration and then fractionated by DEAE-cellulose ion exchange chromatography. A fraction eluted from DEAE-cellulose between 0.10-0.25 M sodium chloride contained most of the MAb FH6 reactivity, as shown by antibody affinity chromatography. These results support a hypothesis that high molecular weight glycoproteins produced by colorectal carcinoma tissues are heterogeneous with regard to their carbohydrate chains and their antigenic structures may change during tumor progression.