Identification of T-cell epitopes on the Rhesus polypeptides in autoimmune hemolytic anemia

Robert Norman Barker, Andrew Michael Hall, G R Standen, Jeff Jones, C J Elson

Research output: Contribution to journalArticle

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Abstract

We have shown previously that the Rhesus (Rh) polypeptides are the commonest targets for pathogenic anti-red blood cell (RBC) autoantibodies in patients with autoimmune hemolytic anemia (AIHA). The aim of the current work was to determine whether activated T cells from such patients also mount recall responses to epitopes on these proteins. Two panels of overlapping 15-mer peptides, corresponding to the sequences of the 30-kD Rh proteins associated with expression of the D and Cc/Ee blood group antigens, were synthesized and screened for the ability to stimulate the in vitro proliferation of mononuclear cells from the peripheral blood or spleen of nine AIHA cases. Culture conditions were chosen that favor recall proliferation by previously activated T cells, rather than primary responses. In seven of the patients, including all four cases with autoantibody to the Rh proteins, two or more peptides elicited proliferation, but cells from eight of nine patients with other anemias and seven of nine healthy donors failed to respond to the panels. Multiple peptides were also stimulatory in two positive control donors who had been alloimmunized with Rh D-positive RBCs. Six different profiles of peptides elicited responses in the AIHA patients, and this variation may reflect the different HLA types in the group. Stimulatory peptides were identified throughout domains shared between, or specific to, each of the related 30-kD Rh proteins, but T cells that responded to nonconserved regions did not cross-react with the alternative sequences. Anti-major histocompatibility complex class II antibodies blocked the responses and depletion experiments confirmed that the proliferating mononuclear cells were T cells. Notably, splenic T cells that proliferated against multiple Rh peptides also responded to intact RBCs. We propose that pathogenic autoantibody production in many cases of AIHA is driven by the activation of T-helper cells specific for previously cryptic epitopes on the Rh proteins.
Original languageEnglish
Pages (from-to)2701-2715
Number of pages15
JournalBlood
Volume90
Issue number7
Publication statusPublished - 1 Oct 1997

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Autoimmune Hemolytic Anemia
T-Lymphocyte Epitopes
T-cells
Peptides
T-Lymphocytes
Autoantibodies
Proteins
Epitopes
Blood
Cell Proliferation
Tissue Donors
Cell proliferation
Blood Group Antigens
Helper-Inducer T-Lymphocytes
Major Histocompatibility Complex
Antibody Formation
Anemia
Blood Cells
Spleen
Erythrocytes

Keywords

  • Adult
  • Aged
  • Amino Acid Sequence
  • Anemia
  • Anemia, Hemolytic, Autoimmune
  • Autoantibodies
  • Autoimmune Diseases
  • Cells, Cultured
  • Epitopes
  • Female
  • Humans
  • Immunity, Cellular
  • Immunologic Memory
  • Male
  • Middle Aged
  • Molecular Sequence Data
  • Rh Isoimmunization
  • Rh-Hr Blood-Group System
  • T-Lymphocyte Subsets
  • T-Lymphocytes, Helper-Inducer

Cite this

Barker, R. N., Hall, A. M., Standen, G. R., Jones, J., & Elson, C. J. (1997). Identification of T-cell epitopes on the Rhesus polypeptides in autoimmune hemolytic anemia. Blood, 90(7), 2701-2715.

Identification of T-cell epitopes on the Rhesus polypeptides in autoimmune hemolytic anemia. / Barker, Robert Norman; Hall, Andrew Michael; Standen, G R; Jones, Jeff; Elson, C J.

In: Blood, Vol. 90, No. 7, 01.10.1997, p. 2701-2715.

Research output: Contribution to journalArticle

Barker, RN, Hall, AM, Standen, GR, Jones, J & Elson, CJ 1997, 'Identification of T-cell epitopes on the Rhesus polypeptides in autoimmune hemolytic anemia', Blood, vol. 90, no. 7, pp. 2701-2715.
Barker RN, Hall AM, Standen GR, Jones J, Elson CJ. Identification of T-cell epitopes on the Rhesus polypeptides in autoimmune hemolytic anemia. Blood. 1997 Oct 1;90(7):2701-2715.
Barker, Robert Norman ; Hall, Andrew Michael ; Standen, G R ; Jones, Jeff ; Elson, C J. / Identification of T-cell epitopes on the Rhesus polypeptides in autoimmune hemolytic anemia. In: Blood. 1997 ; Vol. 90, No. 7. pp. 2701-2715.
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abstract = "We have shown previously that the Rhesus (Rh) polypeptides are the commonest targets for pathogenic anti-red blood cell (RBC) autoantibodies in patients with autoimmune hemolytic anemia (AIHA). The aim of the current work was to determine whether activated T cells from such patients also mount recall responses to epitopes on these proteins. Two panels of overlapping 15-mer peptides, corresponding to the sequences of the 30-kD Rh proteins associated with expression of the D and Cc/Ee blood group antigens, were synthesized and screened for the ability to stimulate the in vitro proliferation of mononuclear cells from the peripheral blood or spleen of nine AIHA cases. Culture conditions were chosen that favor recall proliferation by previously activated T cells, rather than primary responses. In seven of the patients, including all four cases with autoantibody to the Rh proteins, two or more peptides elicited proliferation, but cells from eight of nine patients with other anemias and seven of nine healthy donors failed to respond to the panels. Multiple peptides were also stimulatory in two positive control donors who had been alloimmunized with Rh D-positive RBCs. Six different profiles of peptides elicited responses in the AIHA patients, and this variation may reflect the different HLA types in the group. Stimulatory peptides were identified throughout domains shared between, or specific to, each of the related 30-kD Rh proteins, but T cells that responded to nonconserved regions did not cross-react with the alternative sequences. Anti-major histocompatibility complex class II antibodies blocked the responses and depletion experiments confirmed that the proliferating mononuclear cells were T cells. Notably, splenic T cells that proliferated against multiple Rh peptides also responded to intact RBCs. We propose that pathogenic autoantibody production in many cases of AIHA is driven by the activation of T-helper cells specific for previously cryptic epitopes on the Rh proteins.",
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AB - We have shown previously that the Rhesus (Rh) polypeptides are the commonest targets for pathogenic anti-red blood cell (RBC) autoantibodies in patients with autoimmune hemolytic anemia (AIHA). The aim of the current work was to determine whether activated T cells from such patients also mount recall responses to epitopes on these proteins. Two panels of overlapping 15-mer peptides, corresponding to the sequences of the 30-kD Rh proteins associated with expression of the D and Cc/Ee blood group antigens, were synthesized and screened for the ability to stimulate the in vitro proliferation of mononuclear cells from the peripheral blood or spleen of nine AIHA cases. Culture conditions were chosen that favor recall proliferation by previously activated T cells, rather than primary responses. In seven of the patients, including all four cases with autoantibody to the Rh proteins, two or more peptides elicited proliferation, but cells from eight of nine patients with other anemias and seven of nine healthy donors failed to respond to the panels. Multiple peptides were also stimulatory in two positive control donors who had been alloimmunized with Rh D-positive RBCs. Six different profiles of peptides elicited responses in the AIHA patients, and this variation may reflect the different HLA types in the group. Stimulatory peptides were identified throughout domains shared between, or specific to, each of the related 30-kD Rh proteins, but T cells that responded to nonconserved regions did not cross-react with the alternative sequences. Anti-major histocompatibility complex class II antibodies blocked the responses and depletion experiments confirmed that the proliferating mononuclear cells were T cells. Notably, splenic T cells that proliferated against multiple Rh peptides also responded to intact RBCs. We propose that pathogenic autoantibody production in many cases of AIHA is driven by the activation of T-helper cells specific for previously cryptic epitopes on the Rh proteins.

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KW - Anemia, Hemolytic, Autoimmune

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KW - Autoimmune Diseases

KW - Cells, Cultured

KW - Epitopes

KW - Female

KW - Humans

KW - Immunity, Cellular

KW - Immunologic Memory

KW - Male

KW - Middle Aged

KW - Molecular Sequence Data

KW - Rh Isoimmunization

KW - Rh-Hr Blood-Group System

KW - T-Lymphocyte Subsets

KW - T-Lymphocytes, Helper-Inducer

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VL - 90

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EP - 2715

JO - Blood

JF - Blood

SN - 0006-4971

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ER -