Detection of a sulfotransferase (HEC-GlcNAc6ST) in high endothelial venules of lymph nodes and in high endothelial venule-like vessels within ectopic lymphoid aggregates: relationship to the MECA-79 epitope.
The interaction of L-selectin on lymphocytes with sulfated ligands on high endothelial venules (HEVs) of lymph nodes results in lymphocyte rolling and is essential for lymphocyte homing. The MECA-79 monoclonal antibody reports HEV-expressed ligands for L-selectin by recognizing a critical sulfation-dependent determinant on these ligands. HEC-GlcNAc6ST, a HEV-localized sulfotransferase, is essential for the elaboration of functional ligands within lymph nodes, as well as the generation of the MECA-79 epitope. Here, we use an antibody against murine HEC-GlcNAc6ST to study its expression in relationship to the MECA-79 epitope. In lymph nodes, the enzyme is expressed in the Golgi apparatus of high endothelial cells, in close correspondence with luminal staining by MECA-79. In lymph node HEVs of HEC-GlcNAc6ST-null mice, luminal staining by MECA-79 is almost abolished, whereas abluminal staining persists although reduced in intensity. HEV-like vessels in several examples of inflammation-associated lymphoid neogenesis, including nonobese diabetic mice, also exhibit concomitant expression of the sulfotransferase and luminal MECA-79 reactivity. The correlation extends to ectopic lymphoid aggregates within the pancreas of RIP-BLC mice, in which CXCL13 is expressed in islets. Analysis of the progeny of RIP-BLC by HEC-GlcNAc6ST-null mice establishes that the enzyme is responsible for the MECA-79 defined luminal ligands.
Bistrup
Annette
A
Department of Anatomy, University of California, San Francisco, California 94143-0452, USA.
Tsay
Durwin
D
Shenoy
Priti
P
Singer
Mark S
MS
Bangia
Naveen
N
Luther
Sanjiv A
SA
Cyster
Jason G
JG
Ruddle
Nancy H
NH
Rosen
Steven D
SD
D000818:Animals; D000954:Antigens, Surface / Q000737:chemistry* / Q000378:metabolism; D015153:Blotting, Western; D015815:Cell Adhesion Molecules; D018076:DNA, Complementary / Q000378:metabolism; D004727:Endothelium / Q000201:enzymology*; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry; D008024:Ligands; D008198:Lymph Nodes / Q000201:enzymology* / Q000473:pathology; D008214:Lymphocytes / Q000201:enzymology; D008565:Membrane Proteins; D051379:Mice; D008856:Microscopy, Fluorescence; D009088:Mucoproteins; D015238:Sulfotransferases / Q000096:biosynthesis*; D000097811:Carbohydrate Sulfotransferases
D000954:Antigens, Surface; D015815:Cell Adhesion Molecules; D018076:DNA, Complementary; D000939:Epitopes; D008024:Ligands; D008565:Membrane Proteins; D009088:Mucoproteins; D015238:Sulfotransferases; D000097811:Carbohydrate Sulfotransferases; C069886:L-selectin counter-receptors; C489082:Madcam1 protein, mouse