Abstract：
:T cells utilize the vascular adhesion molecule E-selectin to enter inflamed skin. T cells identified by the mAb HECA-452 [cutaneous lymphocyte-associated antigen (CLA) T cells] are enriched in E-selectin ligand expressing cells. However, the proportion of CLA(+) T cells reactive with an E-selectin/Fc chimeric construct, as determined by flow cytometry, can vary considerably between studies. One important variable in these studies has been the E-selectin/Fc chimera used to assess ligand expression. We therefore compared the reactivity of mouse, rat, and human E-selectin/Fc from the same widely used commercial source with peripheral blood CLA(+) CD4(+) T cells, neutrophils, and the promyelocytic cell line HL-60 by flow cytometry and by shear flow assays. We observed that the binding activities of the different E-selectin/Fc chimeras were considerably different. Mouse E-selectin/Fc demonstrated the highest binding activity with human neutrophils and HL-60 cells by both assay approaches, whereas human E-selectin/Fc demonstrated the lowest binding activity. In addition, mouse E-selectin/Fc binding increased essentially in a linear manner with increasing expression of CLA by CD4(+) T cells, whereas human and rat E-selectin/Fc binding occurred with only a subset of CLA(+) CD4(+) T cells. We conclude that there is substantial variability in the reactivity of different E-selectin/Fc constructs, thus caution should be used when assessing E-selectin ligand expression with these reagents. For instance, the discordance in expression of CLA and E-selectin ligands by T cells may in part be due to the E-selectin/Fc construct being used.
journal_name
Immunol Lettjournal_title
Immunology lettersauthors
Ni Z,Walcheck Bdoi
10.1016/j.imlet.2006.12.002subject
Has Abstractpub_date
2007-02-15 00:00:00pages
179-82issue
2eissn
0165-2478issn
1879-0542pii
S0165-2478(06)00279-3journal_volume
108pub_type
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journal_title：Immunology letters
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journal_title：Immunology letters
pub_type： 杂志文章
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journal_title：Immunology letters
pub_type： 杂志文章
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pub_type： 杂志文章
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