Table 1.

Anti-human TSHR antibody production, thyroid function and thyroid pathology in HLA-DR3 transgenic mice after immunization with human TSHR plasmid DNA only or in conjunction with IL-4 or GM-CSF plasmids

Immunogen†	Mouse No.	Sex	TT4 nmol/l‡	TSAbs fmol/ml§	TSBAbs % cAMP release¶	TBII %125I-TSH inhibition††	Flow cytometric analysis‡‡	mTSH mU/l§§	% thyroid infiltration¶¶
Vector + IL-4 control group	19	F	56	760	<30	<10	–	<16	0
	20	F	57	860	<30	<10	–	<16	0
	21	F	63	400	<30	<10	–	<16	>0–10
	22	M	66	900	<30	<10	–	31	0
	23	M	46	1500	<30	<10	–	52	0
Mean + 3SD	–		81	2080					–
TSHR	1	F	59	1240	44	43*	+ +	<16	0
	2	F	81	1130	<30	11	–	<16	0
	3	F	15	1110	<30	<10	+	<16	0
	4	M	96	4080	<30**	36*	+ +	48	>10–20
	5	M	85	6360*	<30	<10	–	36	0
	6	M	76	2440*	<30	<10	–	24	0
TSHR + IL-4	7	F	69	1160	<30	<10	+ +	ND	0
	9	F	86	1400	<30	<10	+ +	<16	>0–10
	10	F	62	960	<30	<10	+ +	ND	>0–10
	11	M	67	1500	<30	<10	+ +	ND	0
	12	M	62	1570	<30	<10	–	ND	>20–40
TSHR + GM-CSF	13	F	52	460	<30	<10	+ +	ND	0
	14	F	78	680	<30	<10	+	ND	0
	15	F	76	1240	<30	<10	+	ND	0
	16	F	79	840	<30	28	+	<16	0
	17	M	59	1720	<30	<10	+	ND	>0–10
	18	M	64	1810	<30	<10	–	60	0

^†HLA-DR3 transgenic Ab°/NOD mice were immunized with 50 µg each of the indicated DNA plasmids on weeks 0, 3 and 6. Sera were collected on weeks 8, 11 and 14 (when mice were sacrificed). The presented data were derived from week 14. All values considered positive were represented in bold.

^‡Total T₄ assessed using a RIA kit. Values above mean + 3SD of control group were considered positive.

^§Thyroid-stimulating Abs were measured by cAMP release from human TSHR⁺ JP09 cells after 4 h. Values above mean + 3SD of control group were considered positive;

^*denotes week 8 and 11 sera also positive.

^¶Thyroid-stimulating blocking Abs were measured by cAMP release from human TSHR⁺ JP09 cells in the presence of 40 µU/ml bTSH. Values ≥30% were considered positive;

^**denotes week 8 and 11 sera positive.

^††TSH-binding inhibitory immunoglobulins were determined with TRAK II kits. Values ≥10% of control group were considered positive;

^*denotes week 8 and 11 sera also positive.

^‡‡Serum-labelled GPI9-5 cells expressing human TSHR-ECD. Negative (–), positive (+) or strongly positive (+ +) denote fluorescent intensity.

^§§Mouse TSH serum level determined by double-antibody precipitation RIA.

^¶¶Extent of mononuclear cell infiltration determined from 50 to 60 sections of thyroid gland. ND, not determined.