Table 3. Sixteen nonamers predicted from... | American Society of Hematology

Table 3.

Sixteen nonamers predicted from the results of positional scanning synthetic combinatorial library screen

Position		P1	P2	P3	P4	P5	P6	P7	P8	P9
Amino acid		Y	L	L	P	M	V	V	S	V
					W	Y		T	P
Peptide name	Peptide sequence										Concentration, nM
44	H-	Y	L	L	P	M	V	V	S	V	0.00005
46	H-	Y	L	L	P	M	V	T	S	V	0.0005
54	H-	Y	L	L	W	M	V	T	S	V	0.5
56	H-	Y	L	L	W	Y	V	V	S	V	50
47	H-	Y	L	L	P	M	V	T	P	V	500
52	H-	Y	L	L	W	M	V	V	S	V	500
53	H-	Y	L	L	W	M	V	V	P	V	500
55	H-	Y	L	L	W	M	V	T	P	V	500
58	H-	Y	L	L	W	Y	V	T	S	V	500
45	H-	Y	L	L	P	M	V	V	P	V	> 500
48	H-	Y	L	L	P	Y	V	V	S	V	> 500
49	H-	Y	L	L	P	Y	V	V	P	V	> 500
50	H-	Y	L	L	P	Y	V	T	S	V	> 500
51	H-	Y	L	L	P	Y	V	T	P	V	> 500
57	H-	Y	L	L	W	Y	V	V	P	V	> 500
59	H-	Y	L	L	W	Y	V	T	P	V	> 500
pp65_495-503	H-	N	L	V	P	M	V	A	T	V	0.5
pp65_495-503	H-	N	L	V	P	M	V	A	T	V	0.5

Position		P1	P2	P3	P4	P5	P6	P7	P8	P9
Amino acid		Y	L	L	P	M	V	V	S	V
					W	Y		T	P
Peptide name	Peptide sequence										Concentration, nM
44	H-	Y	L	L	P	M	V	V	S	V	0.00005
46	H-	Y	L	L	P	M	V	T	S	V	0.0005
54	H-	Y	L	L	W	M	V	T	S	V	0.5
56	H-	Y	L	L	W	Y	V	V	S	V	50
47	H-	Y	L	L	P	M	V	T	P	V	500
52	H-	Y	L	L	W	M	V	V	S	V	500
53	H-	Y	L	L	W	M	V	V	P	V	500
55	H-	Y	L	L	W	M	V	T	P	V	500
58	H-	Y	L	L	W	Y	V	T	S	V	500
45	H-	Y	L	L	P	M	V	V	P	V	> 500
48	H-	Y	L	L	P	Y	V	V	S	V	> 500
49	H-	Y	L	L	P	Y	V	V	P	V	> 500
50	H-	Y	L	L	P	Y	V	T	S	V	> 500
51	H-	Y	L	L	P	Y	V	T	P	V	> 500
57	H-	Y	L	L	W	Y	V	V	P	V	> 500
59	H-	Y	L	L	W	Y	V	T	P	V	> 500
pp65_495-503	H-	N	L	V	P	M	V	A	T	V	0.5
pp65_495-503	H-	N	L	V	P	M	V	A	T	V	0.5

Nonamer peptides with the substitutions as shown at each position were synthesized as described in “Materials and methods.” Substitutions predicted by the library screen of TCC 3-3F4 are shown on top of the table, immediately below each residue P of the CTL epitope, and the native sequence of the pp65_495-503 CTL epitope is shown at the bottom of the table. Results of CRA are shown using peptide-sensitized HLA-A

F3-150

0201–restricted EBVLCLs as target cells and TCC 3-3F4 effectors at an E/T ratio of 5. The terminus for each peptide and for pp65_495-503 is −NH2, and the terminus for pp65_495-503 is −OH. The concentration of peptide, in which specific killing was between 15% and 30%, represents the average results from at least 3 independent experiments carried out on different days using different batches of TCC-3F4. Peptides were tested using 10-fold dilutions ranging from 500-0.00005 nM.

CTL indicates cytotoxic T-lymphocyte; CRA, chromium release assay; EBVLCLs, Epstein-Barr virus–transformed B cell lines; E/T, effector-to-target; for other abbreviations, see Table 1.

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