(A) Bulk CD8+ T cells from healthy individuals showed
an increase in proliferation upon anti-CD3/28 stimulation and additional
2B4 cross-linking as compared to anti-CD3/28 stimulation alone. This,
however, was only the case for cells with a low 2B4 expression level
ex vivo (left plots), while cells with high 2B4
expression level ex vivo could not be further enhanced
by 2B4 cross-linking (right plots). Ex vivo 2B4
expression levels are indicated above the respective plot. Frequencies
of CFSE-low CD8+ T cells are given, representative plots are shown.
(B) Additionally, the anti-CD3/28-induced degranulation
(as analyzed by CD107a/b expression, left plot) and the IFNγ
production (right plot) of bulk CD8+ T cells from healthy
individuals with low ex vivo 2B4 expression levels
could be enhanced by 2B4 cross-linking in several individuals.
Stimulation indices (SI) are shown referring to anti-CD3/28 stimulation
alone. (C) Expansion of antigen-specific CD8+ T cells
upon peptide-specific stimulation could not be enhanced by additional
2B4-cross-linking in the case of (2B4 high expressing) CMV- or
EBV-specific CD8+ T cells (left plot). However, expansion of (low
2B4 expressing) Flu-specific T cells increased upon additional 2B4
cross-linking in most individuals analyzed (right plot). Stimulation
indices are shown referring to peptide stimulation alone.
(D) Expansion of Flu-specific CD8+ T cells induced
by peptide stimulation and additional 2B4 cross-linking is due to an
increased proliferation of antigen-specific T cells as seen by a higher
frequency of CFSE-low cells. In contrast, CMV- or EBV-specific T cells
did not respond to additional 2B4 cross-linking. (E)
Degranulation (squares) and IFNγ production (triangles) of CMV- or
EBV-specific T cells from healthy individuals did not increase upon 2B4
cross-linking in addition to peptide stimulation (upper panels). In
contrast, these effector functions increased upon peptide stimulation
and simultaneous 2B4 cross-linking in the case of Flu-specific T cells
(lower panels).