A GM-CSF-neuroantigen tolerogenic vaccine elicits inefficient antigen recognition events below the CD40L triggering threshold to expand CD4+ CD25+ FOXP3+ Tregs that inhibit experimental autoimmune encephalomyelitis (EAE)

Table 1 The tolerogenic activity of GMCSF-MOG was commensurate with levels of pre-existing Tregs

Exp.	Strain	Induction of EAE	Group	Treatment	Incidence of EAE	Mean cumulative score	Mean maximal score	% Maximal weight loss
1	C57BL/6	Active	a	GMCSF-MOG	2 of 8	2.1 ± 1.5	0.3 ± 0.2	− 0.8 ± 2.1
1	C57BL/6	Active	b	GMCSF-OVA	7 of 7	81.1 ± 6.9	3.9 ± 0.1	17.4 ± 4.1
2	2D2-FIG-Rag1^−/−	Vaccine	c	GMCSF-MOG	3 of 6	49.0 ± 22.4	1.7 ± 0.8	6.4 ± 4.3
2	2D2-FIG-Rag1^−/−	Vaccine	d	GMCSF-NFM	5 of 6	48.3 ± 16.9	2.6 ± 0.5	4.4 ± 1.6
2	2D2-FIG-Rag1^−/−	Vaccine	e	Saline	0 of 5	0.0 ± 0.0	0.0 ± 0.0	− 2.7 ± 0.9
2	2D2-FIG	Vaccine	f	GMCSF-MOG	0 of 6	0.0 ± 0.0	0.0 ± 0.0	− 4.0 ± 2.8
2	2D2-FIG	Vaccine	g	GMCSF-NFM	0 of 6	0.0 ± 0.0	0.0 ± 0.0	− 2.4 ± 1.8
2	2D2-FIG	Vaccine	h	GMCSF-MOG + GMCSF-NFM	0 of 6	0.0 ± 0.0	0.0 ± 0.0	− 0.7 ± 2.2
2	2D2-FIG	Vaccine	i	Saline	0 of 6	0.0 ± 0.0	0.0 ± 0.0	− 1.6 ± 0.9

Experiment 1: C57BL/6 mice were treated with 2 nmol GMCSF-MOG (n = 8) or GMCSF-OVA (n = 7) on days − 21, − 14, and − 7. On day 0, all mice were immunized with 200 μg MOG^35–55 in CFA and 400 ng of Pertussis toxin i.p. on days 0 and 2. The % maximum weight loss between days 1 and 40 was analyzed with a two-tailed Student’s t test. (a) versus (b), (p ≤ 0.001). Negative values reflect weight gain. Mean cumulative and maximal scores were analyzed with a Mann-Whitney U test (a) versus (b), (p ≤ 0.001)
Experiment 2: 2D2-FIG (n = 6) or 2D2-FIG-Rag1^−/− (n = 5–6) mice were vaccinated with 4 nmol GMCSF-MOG, GMCSF-NFM, “GMCSF-MOG + GMCSF-NFM”, or with saline on days 0 and day 20. The mean cumulative and maximal scores were analyzed with a Kruskal-Wallis Test and adjusted by the Bonferroni correction. Mean cumulative scores (d) versus (e–i), (p < 0.01) and mean maximal scores (c) versus (f–i) (p < 0.05) and (d) versus (e–i), (p < 0.01)

ISSN: 1742-2094