Epitope mapping, expression and post-translational modifications of two isoforms of CD33 (CD33M and CD33m) on lymphoid and myeloid human cells.

Pérez Oliva, Ana B.; Vicente Fernández, José J.; Corral San Miguel, Rubén; García Peñarrubia, Pilar; Hernández Caselles, Trinidad; Martínez-Esparza Alvargonzález, María Concepción

Publication:
Epitope mapping, expression and post-translational modifications of two isoforms of CD33 (CD33M and CD33m) on lymphoid and myeloid human cells.

dc.contributor.author	Pérez Oliva, Ana B.
dc.contributor.author	Vicente Fernández, José J.
dc.contributor.author	Corral San Miguel, Rubén
dc.contributor.author	García Peñarrubia, Pilar
dc.contributor.author	Hernández Caselles, Trinidad
dc.contributor.author	Martínez-Esparza Alvargonzález, María Concepción
dc.contributor.department	Bioquímica y Biología Molecular B e Inmunología
dc.date.accessioned	2024-07-03T07:51:16Z
dc.date.available	2024-07-03T07:51:16Z
dc.date.issued	2011-01-28
dc.description	© The Author 2011. This document is the Published version of a Published Work that appeared in final form in Glycobiology. To access the final edited and published work see https://doi.org/10.1093/glycob/cwq220
dc.description.abstract	We have tested the usefulness of several commercial anti-CD33 monoclonal antibodies (mAb) to determine the expression and localization of the two CD33 isoforms on several hematopoietic cell lines. The expression of the isoform CD33m, a CD33 transmembrane splice variant lacking the ligand-binding V immunoglobulin (Ig)-like domain, was detected by RT-polymerase chain reaction, western blot, confocal microscopy and flow cytometry on the membrane of several human cell types. CD33m was only detected by the anti-CD33 mAb HIM3-4 on the cell surface, whereas WM53, P67.6, 4D3, HIM3-4, WM54, D3HL60.251 or MY9 detected the CD33M isoform, indicating that HIM3-4 is the only mAb recognizing CD33 C(2) Ig domain. Accordingly, HIM3-4 binding to CD33 did not interfere with the binding of other antibodies against the CD33 V-domain. P67.6 mAb interfered with recognition by the rest of antibodies specific for the V domain. HIM3-4 staining could be increased after the sialidase treatment of all CD33(+) cells. However, this increase was stronger in activated T cells, suggesting a CD33 masking state in this cell population. Confocal microscopy analysis of CD33m HEK 293T-transfected cells revealed that this protein is expressed on the cell membrane and also detected in the Golgi compartment. CD33 is constitutively located outside the lipid raft domains, whereas cross-linked CD33 is highly recruited to this signaling platform. The unique ability of HIM3-4 mAb to detect the masking state of CD33 on different cell lineages makes it a good tool to improve the knowledge of the biological role of this sialic acid-binding Ig-like lectin.	es
dc.format	application/pdf	es
dc.format.extent	14	es
dc.identifier.citation	Glycobiology, 2011, Vol. 21, Issue 6, pp. 757–770
dc.identifier.doi	https://doi.org/10.1093/glycob/cwq220
dc.identifier.issn	Print: 0959-6658
dc.identifier.issn	Electronic: 1460-2423
dc.identifier.uri	http://hdl.handle.net/10201/142811
dc.language	eng	es
dc.publisher	Oxford University Press	es
dc.relation	This work was supported by projects 03112/PI/05 and 11926/ PI/09 from Fundación Séneca (CARM, Spain).	es
dc.relation.publisherversion	https://academic.oup.com/glycob/article/21/6/757/1988816	es
dc.rights.accessRights	info:eu-repo/semantics/restrictedAccess
dc.subject	Anti-CD33 mAb	es
dc.subject	CD33 epitopes	es
dc.subject	CD33m isoform	es
dc.title	Epitope mapping, expression and post-translational modifications of two isoforms of CD33 (CD33M and CD33m) on lymphoid and myeloid human cells.	es
dc.type	info:eu-repo/semantics/article	es
dspace.entity.type	Publication	es
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