Supplementary Figure 1: Long term appearance of accepted lung grafts. (a) Gross
appearance
of
Balb/c
lung
allograft
13
months
after
transplantation
into
immunosuppressed (MR1, CTLA4-Ig) B6 recipient (Tx depicts transplanted graft). (b)
Histological appearance of native B6 right lung and (c) BALT in Balb/c allograft 7
months after transplantation into immunosuppressed (MR1, CTLA4-Ig) B6 recipient
(100x). (d) represents a zoomed view of BALT shown in (c) (200x).
Supplementary Figure 2: Appearance of immunosuppressed heart grafts. Two-photon
image of dendritic cells in (a) heart explants derived from resting B6 CD11c+ EYFP mice
and (b) Balb/c heart graft explants 30 days after transplantation into immunosuppressed
(MR1, CTLA4-Ig) B6 CD11c+ EYFP recipients. Scale bar = 20 µm. (c) Immunostaining
(100x)
for
PNAd
for
B6
heart
grafts
30
days
after
transplantation
into
immunosuppressed (MR1, CTLA4-Ig) CBA hosts.
Supplementary Figure 3: PD-L1 expression on dendritic cells in BALT of accepted lung
allografts. (a) Flow cytometric analysis of PD-L1 expression on myeloid dendritic cells
(CD11c+CD11bhi) in accepted B6 lung allografts 30 days after transplantation into
immunosuppressed (MR1, CTLA4-Ig) CBA recipients (n=3) Shaded histogram depicts
staining with isotype control antibody. (b) Immunofluorescent staining showing
expression of PD-L1 (red) on CD11c+ dendritic cells (green) within BALT of accepted
Balb/c lung allografts 30 days after transplantation into immunosuppressed (MR1,
CTLA4-Ig) B6 CD11c-EYFP hosts.
1
Supplementary Movie 1: Intravital two-photon imaging of dendritic cells (green) within
BALT
in
accepted
Balb/c
lung
grafts
30
days
after
transplantation
into
immunosuppressed (MR1 250 µg i.p. day 0, CTLA4-Ig 200 µg i.p. day 2) B6 CD11c+
EYFP recipients. Relative time is displayed in hrs:min:sec:msec.
Supplementary Movie 2:
Intravital two-photon imaging of adoptively transferred
CMTMR-labeled B6 T lymphocytes (yellow) with dendritic cells (green) within BALT in
accepted Balb/c lung grafts 30 days after transplantation into immunosuppressed (MR1
250 µg i.p. day 0, CTLA4-Ig 200 µg i.p. day 2) B6 CD11c+ EYFP recipients. Collagen
and alveolar macrophages appear blue (second harmonic generation) and blood
vessels
appear
red.
Relative
time
2
is
displayed
in
hrs:min:sec:msec.
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