Table 3 Antagonist and agonist activity of double substituted or mixtures of single substituted variants of p57–73 QE65.

Double substitution	Antagonism (%) [p value]	Agonism (%)	Single substitution combination	Antagonism (%) [p value]	*Agonism (%)
64P→W,65E→H	20 (8) [0.02]	1 (4)	64P→W+65E→H	28 (5) [0.00008]	25 (11)
64P→W,65E→I	28 (6) [0.0003]	2 (5)	64P→W+65E→I	28 (5) [0.0003]	26 (10)
64P→W,65E→R	13 (9) NS	−1 (2)	64P→W+65E→R	26 (9) [0.01]	3 (3)
64P→W,65E→T	28 (5) [0.0004]	−1 (3)	64P→W+65E→T	30 (4) [0.00003]	20 (7)
64P→W,65E→W	12 (6) [0.04]	1 (4)	64P→W+65E→W	32 (8) [0.001]	16 (6)
64P→W,67P→I	29 (5) [0.0002]	5 (8)	64P→W+67P→I	22 (7) [0.006]	25 (6)
64P→Y,65E→H	7 (7) NS	7 (10)	64P→Y+65E→H	17 (6) [0.008]	22 (7)
64P→Y,65E→I	9 (5) [0.049]	2 (5)	64P→Y+65E→I	25 (5) [0.0003]	25 (4)
64P→Y,65E→R	4 (7) NS	2 (5)	64P→Y+65E→R	26 (5) [0.0003]	7 (2)
64P→Y,65E→T	18 (5) [0.003]	12 (15)	64P→Y+65E→T	13 (5) [0.01]	17 (7)
64P→Y,65E→W	10 (4) NS	3 (6)	64P→Y+65E→W	32 (4) [0.00001]	7 (1)
64P→Y,67P→I	11 (5) [0.04]	2 (5)	64P→Y+67P→I	15 (8) [0.05]	22 (5)
65E→H,67P→I	−2 (5) NS	3 (6)	65E→H+67P→I	12 (5) [0.02]	31 (5)
65E→I,67P→I	−10 (4) NS	4 (7)	65E→I+67P→I	11 (5) [0.02]	17 (6)
65E→R,67P→I	−3 (5) NS	2 (5)	65E→R+67P→I	12 (6) [0.04]	17 (3)
65E→T,67P→I	−1 (5) NS	3 (6)	65E→T+67P→I	6 (6) NS	16 (5)
65E→W,67P→I	−7 (3) NS	1 (4)	65E→W+67P→I	18 (5) [0.002]	11 (6)

NS, not significant by paired Student's t test.

Antagonism is per cent reduction in interferon γ (IFN‐γ) ELISPOT responses to p57–73 QE65 (10 μg/ml) alone compared with coincubation with substituted peptide (50 μg/ml).

Agonism is IFN‐γ ELISPOT response to double‐substituted peptide (50 μg/ml) or two single substituted peptides (each 50 μg/ml) compared with p57–73 QE65 (50 μg/ml). (*Agonism of single substituted peptide combinations was assessed in eight subjects separately from other assays.)

Values are mean (SEM); n = 10 subjects.