PMID- 33208049
OWN - NLM
STAT- MEDLINE
DCOM- 20220119
LR  - 20220119
IS  - 1557-9077 (Electronic)
IS  - 1050-7256 (Linking)
VI  - 31
IP  - 6
DP  - 2021 Jun
TI  - Thyrotropin Receptor-Specific Lymphocytes in Adenovirus-TSHR-Immunized Native and 
      Human Leukocyte Antigen-DR3-Transgenic Mice and in Graves' Disease Patient Blood.
PG  - 950-963
LID - 10.1089/thy.2020.0338 [doi]
AB  - Background: Antigen-specific lymphocytes are increasingly investigated in 
      autoimmune diseases and immune therapies. We sought to identify thyrotropin 
      receptor (TSHR)-specific lymphocytes in mouse models of Graves' disease, 
      including Graves' patient-specific immunotype human leukocyte antigen (HLA)-DR3, 
      and in frozen and thawed Graves' patient blood samples. Methods and Results: 
      Splenic lymphocytes of adenovirus (Ad)-TSHR-immunized BALB/c mice were stimulated 
      with TSHR-specific peptides C, D, or J. Furthermore, CD154-expressing cells were 
      enriched, expanded in vitro, and analyzed for binding of peptide-major 
      histocompatibility complex (MHC) II multimers ("tetramers," immunotype H2-IA(d)). 
      Only peptides C and J were able to elicit increased expression/secretion of CD154 
      and interferon-gamma, and tetramers which were loaded with peptide C resulted in 
      antigen-specific signals in splenic lymphocytes from Ad-TSHR-immunized mice. 
      Accordingly, TSHR-specific HLA-DR3-MHC class II tetramers loaded with peptide p10 
      specifically bound to human HLA-DR3-(allele B1*03:01)-transgenic Bl/6 mouse 
      splenic T lymphocytes. In addition, we fine-tuned a protocol to reliably measure 
      thawed human peripheral blood mononuclear cells (PBMCs), which resulted in 
      reliable recovery after freezing and thawing with regard to vitality and B and T 
      cell subpopulation markers including regulatory T cells (CD3, CD4, CD25, FoxP3, 
      CD25(high), CD127(low)). TSHR-specific HLA-DR3-MHC class II tetramers loaded with 
      peptide p10 identified antigen-specific T cells in HLA-DR3-positive Graves' 
      patients' thawed PBMCs. Moreover, stimulation-dependent release of interleukin 
      (IL)-1beta, IL-6, tumor necrosis factor-alpha from thawed PBMCs occurred at the 
      expected levels. Conclusions: Novel MHC II tetramers identified TSHR-specific T 
      lymphocytes in Ad-TSHR-immunized hyperthyroid BALB/c or HLA-DR3-transgenic mice 
      and in thawed human PBMCs from patients with Graves' disease. These assays may 
      contribute to measure both disease severity and effects of novel immune therapies 
      in future animal studies and clinical investigations of Graves' disease.
FAU - Degen, Heidrun
AU  - Degen H
AD  - Procorde-Advancecor, Martinsried, Germany.
FAU - Gavvovidis, Ioannis
AU  - Gavvovidis I
AD  - Max-Delbruck Center for Molecular Medicine in the Helmholtz Association, 
      Berlin-Buch, Germany.
AD  - Department of Immunology, Charite - Universitatsmedizin, Berlin, Germany.
FAU - Blankenstein, Thomas
AU  - Blankenstein T
AD  - Max-Delbruck Center for Molecular Medicine in the Helmholtz Association, 
      Berlin-Buch, Germany.
AD  - Department of Immunology, Charite - Universitatsmedizin, Berlin, Germany.
FAU - Uhland, Kerstin
AU  - Uhland K
AD  - Procorde-Advancecor, Martinsried, Germany.
FAU - Ungerer, Martin
AU  - Ungerer M
AD  - Procorde-Advancecor, Martinsried, Germany.
LA  - eng
PT  - Journal Article
DEP - 20210208
PL  - United States
TA  - Thyroid
JT  - Thyroid : official journal of the American Thyroid Association
JID - 9104317
RN  - 0 (Epitopes)
RN  - 0 (HLA-DR3 Antigen)
RN  - 0 (Histocompatibility Antigens Class II)
RN  - 0 (Interleukin-1beta)
RN  - 0 (Interleukin-6)
RN  - 0 (Peptides)
RN  - 0 (Receptors, Thyrotropin)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - 147205-72-9 (CD40 Ligand)
RN  - 82115-62-6 (Interferon-gamma)
SB  - IM
MH  - Animals
MH  - B-Lymphocytes/immunology
MH  - CD40 Ligand/immunology
MH  - Disease Models, Animal
MH  - Epitopes/immunology
MH  - Graves Disease/*immunology
MH  - HLA-DR3 Antigen/*genetics
MH  - Histocompatibility Antigens Class II/immunology
MH  - Humans
MH  - Hyperthyroidism/*immunology
MH  - Immunization
MH  - Immunophenotyping
MH  - Interferon-gamma/immunology
MH  - Interleukin-1beta/immunology
MH  - Interleukin-6/immunology
MH  - Mice
MH  - Mice, Transgenic
MH  - Peptides
MH  - Receptors, Thyrotropin/*immunology
MH  - T-Lymphocytes, Regulatory/*immunology
MH  - Tumor Necrosis Factor-alpha/immunology
OTO - NOTNLM
OT  - Graves' disease
OT  - HLA-DR3 mice
OT  - T cells
OT  - tetramer assay
EDAT- 2020/11/20 06:00
MHDA- 2022/01/20 06:00
CRDT- 2020/11/19 05:29
PHST- 2020/11/20 06:00 [pubmed]
PHST- 2022/01/20 06:00 [medline]
PHST- 2020/11/19 05:29 [entrez]
AID - 10.1089/thy.2020.0338 [doi]
PST - ppublish
SO  - Thyroid. 2021 Jun;31(6):950-963. doi: 10.1089/thy.2020.0338. Epub 2021 Feb 8.