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Table 1 Characterization of the tolerogenic phenotype of mRNA-electroporated tolDC

From: Clinical and immunological control of experimental autoimmune encephalomyelitis by tolerogenic dendritic cells loaded with MOG-encoding mRNA

 

iDC

mDC

tolDC

tolDC mock EP

tolDC mRNA EP

A. Immune phenotype

Gated on viable cells

CD11c

% positive cells

45.2 (42.9–53.5)

42.1 (27.3–56.2)

73.5 (71.9–76.8)AA,BB

68.7 (66.0–80.1)A,B

68.0 (65.2–73.8)A

CD169

% positive cells

12.0 (6.0–23.6)

22.5 (18.4–28.5)

57.5 (23.4–70.3)A

36.4 (20.5–65.7)

42.5 (19.6–69.5)

F4/80

% positive cells

24.1 (16.4–31.4)

17.3 (11.8–27.6)

76.3 (30.7–82.2)B

70.4 (39.7–82.5)B

75.5 (39.7–82.5)A,B

Gated on viable CD11c+ cells

CD86

MFI

13.9 (9.0–24.9)B

55.1 (37.3–59.4)A

21.4 (16.8–26.5)

37.0 (29.2–52.0)

60.8 (46.6–71.2)AAA,CC

Fold change in MFI compared to iDC

–

3.8 (1.7–4.8)

1.9 (0.8–2.6)D

2.3 (1.6–4.5)

3.3 (2.3–7.1)

MHC-II

MFI

17.3 (9.3–28.8)

19.85 (10.6–27.0)

4.8 (2.1–5.6)A,B

4.7 (2.3–5.3)A,B

3.6 (2.5–5.6)A,B

Fold change in MFI compared to iDC

–

1.2 (0.9–1.5)

0.3 (0.1–0.5)DD,BB

0.3 (0.1–0.4)B

0.3 (0.1–0.4)B

CD40

MFI

2.5 (1.6–2.8)BBB

12.8 (11.0–21.3)AAA

7.5 (5.7–10.1)A

6.0 (4.7–9.4)

7.3 (5.2–9.1)A

Fold change in MFI compared to iDC

–

6.0 (4.4–7.8)

2.8 (2.3–5.0)D,B

2.9 (2.2–3.6)BB

3.2 (2.8–3.9)B

B. Cytokine secretion profile

IL-12p70 (pg/mL)

2.3 (1.5–5.8)BBB

356.3 (193.2–409.6)AAA

73.7 (61.9–132.9)BBB

74.3 (43.2–98.4)BBB

99.0 (48.1–122.3)BBB

  1. A. Flow cytometric analysis of the expression of hallmark DC/macrophage and costimulatory markers by non-electroporated iDC, mDC, tolDC, and mock- or mRNA-electroporated tolDC (n = 8). mRNA-electroporated tolDC were electroporated with eGFP mRNA or full-length MOG mRNA. Results are shown as median (1st quartile-3rd quartile) of percentage of positive cells (compared to isotype control) or of mean fluorescent intensity (MFI). A, statistically significant when compared to iDC; B, statistically significant when compared to mDC; C, statistically significant when compared to tolDC, all using Kruskal-Wallis test with Dunn’s post-hoc test; D, statistically significant when compared to mDC using Mann-Whitney U test; A/B/C/Dp < 0.05; AA/BB/CC/DDp < 0.01; AAA/BBB/CCC/DDDp < 0.001. B. IL-12p70 secretion in the culture supernatant of iDC, mDC, and tolDC (n = 6). For this, DC were electroporated on day 6 of the cell culture protocol and stimulated on day 7 with a maturation cocktail consisting of 1 μg/mL LPS and 1000 IU/mL IFN-γ. Cell culture supernatant was harvested 24 h after addition of the maturation stimulus. Results are shown as mean ± standard deviation; A, statistically significant when compared to iDC; B, statistically significant when compared to mDC, using one-way ANOVA with Bonferonni’s post-hoc test; AAA/BBBp < 0.001