|
|
Blood, Vol. 111, Issue 12, 5621-5628, June 15, 2008
Capture of plasma membrane fragments from target cells by trogocytosis requires signaling in T cells but not in B cells
Blood Aucher et al.
111: 5621
Supplemental materials for: Aucher et al
Files in this Data Supplement:
- Document 1. Supplemental materials and methods (PDF, 71 KB)
- Figure S1. The effect of inhibitors on T- and B-cell activation (JPG, 78.2 KB)
-
DO11.10 CD4+ T cells (left panels), OT-I CD8+ T cells (middle panels) and MD4 B cells (right panels) were exposed to their respective target cells expressing the appropriate antigen (open histograms) or not (closed histograms) in the presence (lower panels) or absence (upper panels) of 10 µM PP2. The histograms presented correspond to the levels of CD69 expression on CD4+, CD8+ or B220+ cells for DO11.10, OT-I and MD4 lymphocytes, respectively. Isotypic controls, which gave signals comparable to those of unstained cells, are represented by dotted lines. Percentages indicate the degree of inhibition of the up-regulation. Similar results were obtained in 5 independent experiments.
- Figure S2. The mutagenised form of membrane-bound HEL is still expressed
-
We had two mAbs recognising HEL at our disposal: F10.6.6 is insensitive to the K97A mutation, whereas F9.13.7 recognizes the same epitope as the MD4 BCR on HEL and has about a 300-fold lower binding affinity for the K97A mutant than for WT HEL8. Curves show the specific binding of F10.6.6 (A) and F9.13.7 (B), to various target cells: HEK cells (circles), HEK cells expressing membrane-bound WT HEL (HEKmHELWT; squares), and HEK cells expressing membrane-bound mutant HEL (HEKmHELK97A; triangles). Target cells were incubated with the indicated concentrations of the relevant biotinylated mAb in the absence (total binding) or presence (non specific binding) of 1 µM of the same unlabelled mAb. After three washes, cells were then stained with fluorescent streptavidin. Specific binding to HEL was defined as the difference between the total binding and the non-specific binding (which was negligible, not shown).
|
|
