Identification of Candidate Tolerogenic CD8⁺ T Cell Epitopes for Therapy of Type 1 Diabetes in the NOD Mouse Model

Table 2

Summary of peptide epitope data. Cells were obtained from mice at ages as indicated and were assayed as described in “Section 2.” For stimulation of proliferation the data was internally controlled for each experiment and scored as “fold stimulation” (over background using control peptide) as determined as described in Section 2. Examples of each type of data are shown in the paper figures.

(a)  

Antigen	aa	Candidate peptides that stimulate			Candidate peptides that stimulate
		pancreatic LN cell proliferation			pancreatic LN cell secretion of IFNγ
		3 weeks	13 weeks	Diabetic	3 weeks	13 weeks	Diabetic
Control	147–155	−	−	−	−	−	−
Insulin	39–47	−	+	+	+	+	+
Gad65	178–186	−	+	−	−	−	−
Gad65	546–554	−	+	+	−	+	+
IGRP	206–214	−	+	+	−	+	+
ICAp69	78–86	−	−	+	−	−	−
ZnT8	158–166	−	+	+	−	−	−
ZnT8	282–290	−	+	+	−	+	+
DβH	233–241	−	+	+	−	+	+

(b)  

Antigen	RMAS	Tetramer binding			Tolerance induction
		3 weeks	13 weeks	Diabetic	Purified peptide	Peptide + Zymosan A	Plasmid
Control	+	−	−	0.04
Insulin	+	0.03	0.72	0.89	nt	+	+
Gad65	−	0.02	0.1	0.1	nt	nt	nt
Gad65	+	0.03	0.39	0.14	nt	ns	nt
IGRP	+	−	0.63	0.57	nt	ns	nt
ICAp69	+	−	−	−	nt	ns	nt
ZnT8158	+	−	0.20	0.26	+	ns	nt
ZnT8282	+	−	0.69	0.60	+	ns	ns
DβH	+	0.09	0.41	0.32	+	nt	ns
Ins + DβH						nt	+
Ins + ZnT8						nt	+
Ins + DβH + ZnT8						nt	+

Control peptide is influenza NP.
“−” and “+” refer to whether peptide surpasses cutoff assignment.
“nt”: not tested.
“ns”: not significant.
a: scoring cutoff is >5-fold over control.
b: scoring cutoff is >500 pg/mL. (control is <100 pg/mL).
c: scoring cutoff is >2-fold difference in MFI.
d: % of pancreatic LN CD8⁺ T cells.
e: “+” indicates scoring cutoff has statistically significant value versus control peptide.