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Table 2 T hybridoma response to single site (alanine or aspartic acid) substituted altered peptide ligands of HC gp-39(263–275)

From: Identification of an altered peptide ligand based on the endogenously presented, rheumatoid arthritis-associated, human cartilage glycoprotein-39(263–275) epitope: an MHC anchor variant peptide for immune modulation

    

Hybridoma response (SI)

Compound

HC gp-39

Sequence

HLA-DR binding IC50 (μM)

5G11

8B12

14G11

20H5

1

263–275

RSFTLASSETGVG

0.08

17

39

30

5

10

R263A1

ASFTLASSETGVG

0.10

23

42

28

5

11

S264A2

RAFTLASSETGVG

0.10

21

43

32

5

12

F265A3

RSATLASSETGVG

>35

2

6

2

4

13

T266A4

RSFALASSETGVG

0.06

7

14

1

5

14

L267A5

RSFTAASSETGVG

0.08

4

1

1

4

15

A268D6

RSFTLDSSETGVG

0.24

1

1

1

2

16

S269A7

RSFTLAASETGVG

0.05

2

1

1

2

17

S270A8

RSFTLASAETGVG

0.05

21

29

27

5

18

E271A9

RSFTLASSATGVG

0.04

1

1

1

2

19

T272A10

RSFTLASSEAGVG

0.04

1

4

1

1

20

G273A11

RSFTLASSETAVG

0.05

7

32

6

5

21

V274A12

RSFTLASSETGAG

0.06

26

40

30

5

22

G275A13

RSFTLASSETGVA

0.05

17

30

20

5

23

A3/A8

RSATLASAETGVG

>36

ND

ND

ND

ND

  1. Binding of peptides to HLA-DR4 (B1*0401) was determined in a competition binding assay. IC50 = 50% inhibitory concentration. IC50 < 1 μM = high affinity binder; IC50 between 1 and 10 μM = good binder; IC50 between 10 and 100 μM = intermediate binder. The HLA-DRB1*0401 expressing B lymphoblastoid cell line was used as a source of antigen-presenting cells. Background values for hybridomas 5G11, 8B12, 14G11 and 20H5 were 17,238, 8,773, 10,131 and 290,666 mean fluorescence units/counts, respectively. The standard deviation of measurements did not exceed 15%. Hybridoma IL-2 response is expressed as stimulation index (SI) calculated as the ratio of mean fluorescence counts of antigen stimulated cultures and control cultures. Values in bold are considered positive (SI > 2). Data are representative of at least three independent experiments. ND, not determined.
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Arthritis Research & Therapy

ISSN: 1478-6362

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