Systematic determination of TCR-antigen and peptide-MHC binding kinetics among field variants of a Theileria parva polymorphic CTL epitope

CTLs are known to contribute to immunity toward Theileria parva, the causative agent of East Coast fever. The Tp967-75 CTL epitope from the Muguga strain of T. parva is polymorphic in other parasite strains. Identifying the amino acids important for MHC class I binding, as well as TCR recognition of...

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Main Authors: Svitek, Nicholas, Saya, Rosemary, Houshuang Zhang, Nene, Vishvanath M., Steinaa, Lucilla
Format: Journal Article
Language:Inglés
Published: Oxford University Press 2022
Subjects:
Online Access:https://hdl.handle.net/10568/117535
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author Svitek, Nicholas
Saya, Rosemary
Houshuang Zhang
Nene, Vishvanath M.
Steinaa, Lucilla
author_browse Houshuang Zhang
Nene, Vishvanath M.
Saya, Rosemary
Steinaa, Lucilla
Svitek, Nicholas
author_facet Svitek, Nicholas
Saya, Rosemary
Houshuang Zhang
Nene, Vishvanath M.
Steinaa, Lucilla
author_sort Svitek, Nicholas
collection Repository of Agricultural Research Outputs (CGSpace)
description CTLs are known to contribute to immunity toward Theileria parva, the causative agent of East Coast fever. The Tp967-75 CTL epitope from the Muguga strain of T. parva is polymorphic in other parasite strains. Identifying the amino acids important for MHC class I binding, as well as TCR recognition of epitopes, can allow the strategic selection of Ags to induce cellular immunity toward T. parva In this study, we characterized the amino acids important for MHC class I binding and TCR recognition in the Tp967-75 epitope using alanine scanning and a series of variant peptide sequences to probe these interactions. In a peptide-MHC class I binding assay, we found that the amino acids at positions 1, 2, and 3 were critical for binding to its restricting MHC class I molecule BoLA-1*023:01. With IFN-γ ELISPOT and peptide-MHC class I Tet staining assays on two parasite-specific bovine CTL lines, we showed that amino acids at positions 5-8 in the epitope were required for TCR recognition. Only two of eight naturally occurring polymorphic Tp9 epitopes were recognized by both CTLs. Finally, using a TCR avidity assay, we found that a higher TCR avidity was associated with a stronger functional response toward one of two variants recognized by the CTL. These data add to the growing knowledge on the cross-reactivity of epitope-specific CTLs and specificities that may be required in the selection of Ags in the design of a wide-spectrum vaccine for East Coast fever.
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spelling CGSpace1175352025-10-26T13:00:35Z Systematic determination of TCR-antigen and peptide-MHC binding kinetics among field variants of a Theileria parva polymorphic CTL epitope Svitek, Nicholas Saya, Rosemary Houshuang Zhang Nene, Vishvanath M. Steinaa, Lucilla immunology vaccines east coast fever CTLs are known to contribute to immunity toward Theileria parva, the causative agent of East Coast fever. The Tp967-75 CTL epitope from the Muguga strain of T. parva is polymorphic in other parasite strains. Identifying the amino acids important for MHC class I binding, as well as TCR recognition of epitopes, can allow the strategic selection of Ags to induce cellular immunity toward T. parva In this study, we characterized the amino acids important for MHC class I binding and TCR recognition in the Tp967-75 epitope using alanine scanning and a series of variant peptide sequences to probe these interactions. In a peptide-MHC class I binding assay, we found that the amino acids at positions 1, 2, and 3 were critical for binding to its restricting MHC class I molecule BoLA-1*023:01. With IFN-γ ELISPOT and peptide-MHC class I Tet staining assays on two parasite-specific bovine CTL lines, we showed that amino acids at positions 5-8 in the epitope were required for TCR recognition. Only two of eight naturally occurring polymorphic Tp9 epitopes were recognized by both CTLs. Finally, using a TCR avidity assay, we found that a higher TCR avidity was associated with a stronger functional response toward one of two variants recognized by the CTL. These data add to the growing knowledge on the cross-reactivity of epitope-specific CTLs and specificities that may be required in the selection of Ags in the design of a wide-spectrum vaccine for East Coast fever. 2022-02-01 2022-01-17T14:52:15Z 2022-01-17T14:52:15Z Journal Article https://hdl.handle.net/10568/117535 en Open Access Oxford University Press Svitek, N., Saya, R., Zhang, H., Nene, V. and Steinaa, L. 2022. Systematic determination of TCR-antigen and peptide-MHC binding kinetics among field variants of a Theileria parva polymorphic CTL epitope. Journal of Immunology 208(3): 549–561.
spellingShingle immunology
vaccines
east coast fever
Svitek, Nicholas
Saya, Rosemary
Houshuang Zhang
Nene, Vishvanath M.
Steinaa, Lucilla
Systematic determination of TCR-antigen and peptide-MHC binding kinetics among field variants of a Theileria parva polymorphic CTL epitope
title Systematic determination of TCR-antigen and peptide-MHC binding kinetics among field variants of a Theileria parva polymorphic CTL epitope
title_full Systematic determination of TCR-antigen and peptide-MHC binding kinetics among field variants of a Theileria parva polymorphic CTL epitope
title_fullStr Systematic determination of TCR-antigen and peptide-MHC binding kinetics among field variants of a Theileria parva polymorphic CTL epitope
title_full_unstemmed Systematic determination of TCR-antigen and peptide-MHC binding kinetics among field variants of a Theileria parva polymorphic CTL epitope
title_short Systematic determination of TCR-antigen and peptide-MHC binding kinetics among field variants of a Theileria parva polymorphic CTL epitope
title_sort systematic determination of tcr antigen and peptide mhc binding kinetics among field variants of a theileria parva polymorphic ctl epitope
topic immunology
vaccines
east coast fever
url https://hdl.handle.net/10568/117535
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