[en] H2-relaxin (RLN2) is a two-chain peptide hormone structurally related to insulin with a therapeutic potential in multiple indications. However, multiple injections of human RLN2 induced anti-RLN2 Abs in patients, hampering its clinical development. As T cell activation is required to produce Abs, we wondered whether T cells specific for RLN2 might be already present in the human blood before any injection. We therefore quantified the RLN2-specific T cell repertoire using PBMCs collected from healthy donors. CD4 T cells were stimulated in multiple replicates by weekly rounds of stimulation by dendritic cells loaded with RLN2, and their specificity was assessed by IFN-γ ELISPOT. The number of specific T cell lines was used to estimate the frequency of circulating T cells. In vitro T cell response was demonstrated in 18 of the 23 healthy donors, leading to the generation of 70 independent RLN2-specific T cell lines. The mean frequency of RLN2-specific CD4 T cells was similar to that of T cells specific for known immunogenic therapeutic proteins. Using overlapping peptides, we identified multiple T cell epitopes hosted in the N-terminal parts of the α- and β-chains and common to multiple donors, in agreement with their capacity to bind to multiple HLA-DR molecules. Our results provide important clues to the immunogenicity of RLN2 and highlight the weak central immune tolerance induced against this self-hormone.
Disciplines :
Immunology & infectious disease
Author, co-author :
Azam, Aurélien ; Biologics Research, Sanofi Research and Development, 94400 Vitry sur Seine, France ; Commissariat à l'Energie Atomique et aux Energies Alternatives (CEA)-Saclay, Université Paris-Saclay, Service d'Ingénierie Moléculaire des Protéines, 91191 Gif-sur-Yvette, France
Gallais, Yann; Commissariat à l'Energie Atomique et aux Energies Alternatives (CEA)-Saclay, Université Paris-Saclay, Service d'Ingénierie Moléculaire des Protéines, 91191 Gif-sur-Yvette, France
Mallart, Sergio ; Integrated Drug Discovery, Sanofi Research and Development, 91380 Chilly Mazarin, France, and
Illiano, Stephane; Cardiovascular Diseases and Metabolism, Sanofi Research and Development, 91380 Chilly Mazarin, France
Duclos, Olivier ; Integrated Drug Discovery, Sanofi Research and Development, 91380 Chilly Mazarin, France, and
Prades, Catherine; Biologics Research, Sanofi Research and Development, 94400 Vitry sur Seine, France
Maillère, Bernard; Commissariat à l'Energie Atomique et aux Energies Alternatives (CEA)-Saclay, Université Paris-Saclay, Service d'Ingénierie Moléculaire des Protéines, 91191 Gif-sur-Yvette, France, bernard.maillere@cea.fr
Language :
English
Title :
Healthy Donors Exhibit a CD4 T Cell Repertoire Specific to the Immunogenic Human Hormone H2-Relaxin before Injection.
Publication date :
2019
Journal title :
Journal of Immunology
ISSN :
0022-1767
eISSN :
1550-6606
Publisher :
American Association of Immunologists, Bethesda, United States
This work was supported by Sanofi and CEA. This work was also supported by the Laboratory of Excellence in Research on Medication and Therapeutic Innovation (to B.M.).
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