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Supplemental Figure legends following an allogeneic HSCT ... · PDF file Table 1.: Summay of outcomes comparing TrED recipients with PTCy recipients and GVHD controls following an

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Text of Supplemental Figure legends following an allogeneic HSCT ... · PDF file Table 1.: Summay of...

  • 27

    Figure 7.: Significantly higher frequencies of recent thymic/marrow emigrants (RTEs/RMEs) in TrED

    recipients versus PTCy treated animals and GVHD controls early post-HSCT. A HSCT utilizing a B6

    à BALB/c donor/recipient mouse model involving a complete MHC mismatch was performed on day 0.

    Lethally irradiated (8.5 Gy) BALB/c mice received 5x106 TCD B6-RAG2p-GFP BM cells and spleen cells

    from expanded (TL1A-Ig/IL-2; TrED group) or untreated B6-FoxP3rfp (GVHD and PTCy group) donor mice

    adjusted to contain 1.1x106 total T cells. Cyclophosphamide was given on day 3 and 4 post-HSCT at 80

    mg/kg ip. RTEs/RMEs were analyzed in PB by flow cytometry three weeks post HSCT (BM: n = 2; GVHD:

    n = 5; PTCy and TrED n = 8). (A) Significantly higher frequencies of CD4+ and CD8+ RTEs as well as CD19+

    B cells (RMEs) are detected in TrED recipients compared to PTCy treated animals and GVHD controls.

    Data are expressed as means ± SEM and were analyzed by a two-tailed unpaired t test. *p

  • 28

    Supplemental Figure 3.: Treg assessment in blood three and four weeks post-HSCT. (A) Percent

    Foxp3+ Tregs out of total CD4+ T cells (upper graphs) as well as Treg / CD4 ratios (lower graphs) are

    shown for day 21 (left) and 30 (right). Data are shown as mean ± SEM; ANOVA with Bonferroni correction

    was applied for multiple comparisons on day 21. Data are expressed as means ± SEM and were analyzed

    by a two-tailed unpaired t test on day 30. *p

  • 29

    are significant differences detectable in DN3 (CD44-CD25+) and DN4 (CD44-CD25-) subtypes between

    the TrED and the PTCy group (n = 6). Data are expressed as means ± SEM and were analyzed by a

    two-tailed unpaired t test. *p

  • 0

    10

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    % T

    re g

    / C D

    4+

    d d

    0

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    4+

    No n-

    ex pa

    nd ed

    Ex pa

    nd ed No

    n-

    ex pa

    nd ed

    Ex pa

    nd ed

    ******A B

    Supplemental Figure 1: Wolf et al.

  • 6 14 22 30 38 46 54 62 70 78 86 94

    60

    70

    80

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    110

    120

    W ei

    gh t i

    n %

    6 14 22 30 38 46 54 62 70 78 86 94

    0

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    C lin

    ic al

    S co

    re

    0 20 40 60 80 10 0

    12 0

    0

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    Pe rc

    en t s

    ur vi

    va l

    GVHD PTCy TrED

    ns

    Supplemental Figure 2: Wolf et al.

    A B C

    Days post-HSCT Days post-HSCT Days post-HSCT

  • Supplemental Figure 3: Wolf et al. Day 21 Day 30

    GV HD

    PT Cy

    Tr ED

    0

    20

    40

    60

    80

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    120

    Tr eg

    GV HD

    PT Cy

    Tr ED

    0

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    120

    Tr eg

    BM donor T cell donor Recipient

    *

    GV HD

    PT Cy

    Tr ED

    0

    5

    10

    15

    20

    % T

    re g

    / T ot

    al C

    D 4+

    GV HD

    PT Cy

    Tr ED

    0.0

    0.2

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    1.0

    Tr eg

    / To

    ta l C

    D 4+

    ra tio

    GV HD

    PT Cy

    Tr ED

    0

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    % T

    re g

    / T ot

    al C

    D 4+

    GV HD

    PT Cy

    Tr ED

    0.0

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    0.4

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    1.0

    Tr eg

    / To

    ta l C

    D 4+

    ra tio

    ns

    ns ***

    *** ***

    **

    ns

    ns

    A

    B

  • Supplemental Figure 4: Wolf et al.

    GVHD PTCy TrED

    Lu ng

    GV HD

    PT Cy

    Tr ED

    0

    1

    2

    3

    4

    P at

    ho lo

    gi ca

    l S co

    re **

    ****

  • C

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    % C

    D 8

    / L ym

    ph oc

    yt es

    %

    C D

    4 / L

    ym ph

    oc yt

    es

    Day 30 Day 60 Day 90

    Naïve cells (CD62hiCD44lo) Memory cells (CD62loCD44hi) Remaining cells

    Supplemental Figure 5: Wolf et al.

    ***

    ****

    *

    **

    ns

    **

    Stats shown for naïve cells only

    GV HD

    PT Cy

    TrE D

    PT Cy

    TrE D

    PT Cy

    TrE D

  • DN 1 (CD44+/CD25-) DN 2 (CD44+/CD25+) DN 3 (CD44-/CD25+) DN 4 (CD44-/CD25-)

    0

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    D N

    1 - D

    N 4

    *

    ** A B

    Supplemental Figure 6: Wolf et al.

    PT Cy

    TrE D

    PT Cy

    TrE DD

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    120

    D N

    1 - D

    N 4

    ns

    ns

  • BM

    GV HD

    PT Cy

    Tr ED

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    % C

    D 4

    R TE

    s

    BM

    GV HD

    PT Cy

    Tr ED

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    % C

    D 8

    R TE

    s

    BM

    PT Cy

    Tr ED

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    % C

    D 8

    R TE

    s

    CD4 CD8 CD19A

    B

    BM

    PT Cy

    Tr ED

    0

    20

    40

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    80

    % C

    D 4

    R TE

    s

    **

    Supplemental Figure 7: Wolf et al.

    BM

    GV HD

    PT Cy

    Tr ED

    0

    20

    40

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    % C

    D 19

    R M

    E s

    BM

    PT Cy

    Tr ED

    0

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    40

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    % C

    D 19

    R M

    Es

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