Bella Blaher, PhDa, Cenk Suphioglu, PhDb, R. Bruce Knox, DScb, Mohan B

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Presentation transcript:

Identification of T-cell epitopes of Lol p 9, a major allergen of ryegrass (Lolium perenne) pollen Bella Blaher, PhDa, Cenk Suphioglu, PhDb, R.Bruce Knox, DScb, Mohan B. Singh, PhDb, James McCluskey, MB, BSc, Jennifer M. Rolland, PhDa Journal of Allergy and Clinical Immunology Volume 98, Issue 1, Pages 124-132 (July 1996) DOI: 10.1016/S0091-6749(96)70234-8 Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 1 Allergen specificity of the Lol p 9 T-cell line. Cells were cultured with Lol p 9 and crude ryegrass and Bermuda grass pollen extracts for 4 days. Reactivity was assessed by tritiated thymidine incorporation, and the mean cpm ± SD for triplicate wells are shown. The maximum SI response to these allergens was 7.8 for Lol p 9, 3.5 for ryegrass pollen extract, and 1.8 for Bermuda grass pollen extract. The background count in the absence of allergen was 11,300 ± 1246 cpm. [3H]TdR, Tritiated thymidine. Journal of Allergy and Clinical Immunology 1996 98, 124-132DOI: (10.1016/S0091-6749(96)70234-8) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 2 Proliferative response of the Lol p 9 T-cell line to Lol p 9 synthetic peptides. The peptides were tested at 10 μg/ml, and results are expressed as the mean cpm of duplicate cultures. The line responded to Lol p 9 (105-116) (SI = 4.5) and weakly to Lol p 9 (193-204) (SI = 2.3). The mean background count for cells cultured without peptide was 4849 cpm. Journal of Allergy and Clinical Immunology 1996 98, 124-132DOI: (10.1016/S0091-6749(96)70234-8) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 3 Proliferative responses of T-cell clones to the Lol p 9 peptide series (10 μg/ml). Mean cpm of duplicate cultures are shown. A, T-cell clone A12 showed strong reactivity to only Lol p 9 (105-116) (SI = 137). Mean background count was 358 cpm. Note: the y axis is shown on a log scale. B, T-cell clones C2 (filled bars) and C3 (open bars) responded to Lol p 9 (193-204) (SI = 11 and 8.0, respectively) and weakly to the neighboring peptide Lol p 9 (201-212) (SI = 3.2 and 3.0, respectively). Mean background count for T-cell clone C2 was 1063 cpm, and for T-cell clone C3, 2519 cpm. C, T-cell clone A3 showed no response to any of the Lol p 9 peptides. Mean background count was 252 cpm. Journal of Allergy and Clinical Immunology 1996 98, 124-132DOI: (10.1016/S0091-6749(96)70234-8) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 3 Proliferative responses of T-cell clones to the Lol p 9 peptide series (10 μg/ml). Mean cpm of duplicate cultures are shown. A, T-cell clone A12 showed strong reactivity to only Lol p 9 (105-116) (SI = 137). Mean background count was 358 cpm. Note: the y axis is shown on a log scale. B, T-cell clones C2 (filled bars) and C3 (open bars) responded to Lol p 9 (193-204) (SI = 11 and 8.0, respectively) and weakly to the neighboring peptide Lol p 9 (201-212) (SI = 3.2 and 3.0, respectively). Mean background count for T-cell clone C2 was 1063 cpm, and for T-cell clone C3, 2519 cpm. C, T-cell clone A3 showed no response to any of the Lol p 9 peptides. Mean background count was 252 cpm. Journal of Allergy and Clinical Immunology 1996 98, 124-132DOI: (10.1016/S0091-6749(96)70234-8) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 3 Proliferative responses of T-cell clones to the Lol p 9 peptide series (10 μg/ml). Mean cpm of duplicate cultures are shown. A, T-cell clone A12 showed strong reactivity to only Lol p 9 (105-116) (SI = 137). Mean background count was 358 cpm. Note: the y axis is shown on a log scale. B, T-cell clones C2 (filled bars) and C3 (open bars) responded to Lol p 9 (193-204) (SI = 11 and 8.0, respectively) and weakly to the neighboring peptide Lol p 9 (201-212) (SI = 3.2 and 3.0, respectively). Mean background count for T-cell clone C2 was 1063 cpm, and for T-cell clone C3, 2519 cpm. C, T-cell clone A3 showed no response to any of the Lol p 9 peptides. Mean background count was 252 cpm. Journal of Allergy and Clinical Immunology 1996 98, 124-132DOI: (10.1016/S0091-6749(96)70234-8) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 4 Effect of anti-HLA monoclonal antibodies on the response to Lol p 9 (105-116) by T-cell clone A12 (145 days old). T cells were cultured at 1 × 105 cells/ml with 1 × 105 cells/ml irradiated autologous PBMCs and 10 μg/ml Lol p 9 (105-116) in the presence or absence of 5 μl/ml anti-HLA antibody. Proliferation was assessed by the incorporation of tritiated thymidine ([3H]TdR), and results are expressed as mean cpm for triplicate cultures + SD. Mab, Monoclonal antibody. Journal of Allergy and Clinical Immunology 1996 98, 124-132DOI: (10.1016/S0091-6749(96)70234-8) Copyright © 1996 Mosby, Inc. Terms and Conditions