LEGEND TO FIGURES
Figure 1. Effect of CD8 Treg (CD183+CCR7+CD45RA-CD8+) on B
cell proliferation. Purified CFSE labeled B cells were co-cultured in
the presence or absence of various concentrations of CD183+CCR7+CD45RA-
CD8 Treg and activated with anti-CD40 and CpG for 5 days. Cell division
was measured by flow cytometry. (A) shows a representative flow
cytometry histogram. (B) shows cumulative data from 6 different
normal donors as mean ± sem. Inhibition was observed only at 1:1 B:CD8
Treg ratio.
Figure 2. Effect of CD8 Treg (CD183+CCR7+CD45RA-CD8+) on
immunoglobulin production. Purified B cells were co-cultured in the
presence or absence of CD183+CCR7+CD45RA- CD8 Treg at 1:1 ratio and
activated with anti-CD40 and CpG for 14 days. Supernatants were
collected and immunoglobulins were measured by ELISA assay. Data are
expressed for 6 subjects as mean ± sem. CD8 Treg inhibited significantly
IgM ( P<0.005) and IgG (P< 0.039 ) production. No
significant effect was observed on IgA production (P>0.7).
Figure 3. Further characterization of CD8 Treg. Activated CD8+ T cells were stained with various monoclonal antibodies
and isotype controls. FoxP3+ cells were gated and examined for the
expression of different antigens. (A) expression of molecules
on % of CD8+ T cells, and (B) mean flourescence intensity
(MFI) of the different molecules. Data are shown for 5 different
subjects and expressed as mean ± sem. A significant (*) increased
(<P.005) expression of CD25, CD183, CD278, CTLA-4 , and
LAP-1was observed on FoxP3+ CD8+ T cells as compared to FoxP3- cells.
Figure 4. Effect of CD8 Treg
(CD183+CD25highCD278+CD8+) on B cell proliferation.Purified CFSE labeled B cells were co-cultured in the presence or
absence of various concentrations of
CD183+CD25highCD278+ CD8 Treg and activated with
anti-CD40 and CpG for 5 days. Cell division was measured by flow
cytometry. (A) shows a representative flow cytometry histogram.(B) shows cumulative data from 6 different normal donors as
mean ± sem. In a concentration-dependent manner CD8 Treg inhibited B
cell proliferation.
Figure 5. Effect of CD8 Treg
(CD183+CD25highCD278+CD8+) on immunoglobulin
production. Purified B cells were co-cultured in the presence or
absence of CD183+CD25highCD278+ CD8 Treg at various
ratios and activated with anti-CD40 and CpG for 14 days. Supernatants
were collected and immunoglobulins were measured by ELISA assay. Data
are shown from 5 separate subjects (mean ± sem). CD8 Treg markedly
inhibited IgM, IgG, and IgA production at all B cell:CD8 Treg ratios.