Relationship of CD146 expression to activation of circulating T cells: exploratory studies in healthy donors and patients with connective tissue diseases
Version of Record online: 8 SEP 2013
© 2013 British Society for Immunology
Clinical & Experimental Immunology
Volume 174, Issue 1, pages 73–88, October 2013
How to Cite
Hadjinicolaou, A. V., Wu, L., Fang, B., Watson, P. A., Hall, F. C. and Busch, R. (2013), Relationship of CD146 expression to activation of circulating T cells: exploratory studies in healthy donors and patients with connective tissue diseases. Clinical & Experimental Immunology, 174: 73–88. doi: 10.1111/cei.12151
- Issue online: 8 SEP 2013
- Version of Record online: 8 SEP 2013
- Accepted manuscript online: 6 JUN 2013 03:17AM EST
- Manuscript Accepted: 31 MAY 2013
- Pathological Society of Great Britain and Ireland
- Actelion Pharmaceuticals
- Cambridge Biomedical Research Centre of the National Institute for Health Research
- Elmore Fund at Sidney Sussex College and Arthritis Research UK
Fig. S1. Similar patterns of CD146 co-expression with other markers after distinguishing CD3+ T cell subsets by either CD4 or CD8 staining. Peripheral blood mononuclear cells (PBMCs) from a systemic lupus erythematosus (SLE) patient were stained for CD146 and a panel other markers (‘Antigen X’). (a) CD4 T cells were gated either as CD3+CD4+ or CD3+CD8− lymphocytes. Frequencies of CD146+ CD4 cells with or without Antigen X were then enumerated. (b) The same analysis performed for CD8 T cells, which were gated either as CD3+CD4− or CD3+CD8+ lymphocytes. In both subsets, closely similar expression patterns were obtained with either gating procedure.
Fig. S2. No effect of cryopreservation on patterns of CD146 versus CD45RO expression on T cells. Analysis of three systemic lupus erythematosus (SLE) patients. (a) Representative dot-plots from one patient, gated on CD4+ or CD4− T cells. (b) Percentages of indicated subpopulations in three patients. The CD4+/CD4− ratio was also unaffected by cryopreservation.
Fig. S3. Surface CD146 versus intracellular forkhead box protein 3 (FoxP3) expression in gated CD4+ and CD8 peripheral blood T cells from a representative HD (of five analysed).
Fig. S4. CD146 versus human leucocyte antigen D-related (HLA-DQ) [major histocompatibility complex (MHC) class II] expression in peripheral blood CD4+ and CD4− (CD8) T cells from healthy donors (HDs) and connective tissue disease (CTD) patients, analysed as in Fig. 4 of the main paper.
Fig. S5. CD146 versus CD70 expression, analysed as in Fig. 4 of the main paper.
Fig. S6. CD146 versus CD45RA expression in T cells from healthy donors (HDs) and systemic lupus erythematosus (SLE) patients, analysed as in Fig. 4 of the main paper. #Indicates a single donor in whom carryover of CD3− antigen-presenting cells (APC) from an adjacent well caused 100% of T cells to be aberrantly positive for CD146.
Fig. S7. CXCR3 expression in total versus CD146+ CD4 and CD8 T cells from healthy donors (HDs) and systemic lupus erythematosus (SLE) patients; paired analysis as in Fig. 4b,c of the main paper (P > 0·05, not significant).
Fig. S8. CD146 versus CD31 expression, analysed as in Fig. 4 of the main paper.
Fig. S9. CD146 versus CD54/intercellular adhesion molecule 1 (ICAM-1) expression, analysed in healthy donors (HDs) and systemic lupus erythematosus (SLE) patients, as in Fig. 4 of the main paper.
Fig. S10. CD146+ lymphocytes greatly outnumber CD146+ circulating endothelial cells. Peripheral blood mononuclear cells (PBMCs) from a healthy donor were co-stained for CD45 (leucocyte common antigen), CD146 and CD34 (the latter is expressed both on haematopoietic progenitors and on endothelial cells). Numbers represent percentages or frequencies. In the CD45+ leucocyte gate a proportion of cells stained for either CD146 or CD34, but not both. In the CD45− gate, a small number of CD34+CD146+ double-positive events were detected, which may be circulating endothelial cells (versus one event detected in isotype control).
Table S1. Clinical characteristics of patients.
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