Preservation of functioning human thyroid "organoids" in the scid mouse. IV. In vivo selection of an intrathyroidal T cell receptor repertoire.
To study the in vivo influence of thyroid cells on the T cell receptor repertoire in human autoimmune thyroid disease, we mixed lymphocyte-free thyrocytes (approximately 1.2 x 10[6]) from patients with Graves' disease with autologous peripheral blood mononuclear cells (PBMC; approximately 1.5 x 10[6]) and transplanted this mixture sc into scid mice while suspended in a basement membrane gel (approximately 0.4 ml). Controls included mice that received either thyrocytes only or PBMC only. The resulting artificial mixed cell thyroid organoids were explanted after 5 weeks, and their T cell receptor repertoire was examined. Of a total of 63 organoids constructed, 60 were recovered (95.2%). Total RNA was extracted and then analyzed by reverse transcription-PCR primarily for human T cell receptor (hTcR) Vbeta gene expression using 21 hTcR Vbeta amplimers. A restricted pattern of hTcR Vbeta gene expression was found, with 6 Vbeta genes (Vbeta5, 6, 7, 8, 13.1, and 18) predominantly expressed [P < 0.05, by ANOVA on ranks and Student-Newman-Keul's (SNK) test]. PBMC and control organoids showed no preferential selection of particular hTcR V gene-expressing T cells. This reductionist, mixed cell, thyroid model reflected earlier observations in human and murine autoimmune thyroid diseases in which a bias in hTcR V gene family expression had been observed. The model permitted in vivo T cell selection and/or enrichment of potentially disease relevant human T cells.
Martin
A
A
Department of Medicine, Mount Sinai School of Medicine, New York, New York 10029, USA. amartin@smtplink.mssm.edu
Matsuoka
N
N
Zhang
J
J
Zhou
A
A
Nakashima
M
M
Unger
P
P
Schwartz
A E
AE
Friedman
E W
EW
Shultz
L D
LD
Davies
T F
TF
D000818:Animals; D015703:Antigens, CD / Q000494:pharmacology; D000951:Antigens, Neoplasm; D000074301:CD52 Antigen; D015870:Gene Expression; D006023:Glycoproteins; D006111:Graves Disease / Q000097:blood; D006801:Humans; D051379:Mice; D016513:Mice, SCID / Q000502:physiology*; D009000:Monocytes / Q000502:physiology; D009940:Organoids / Q000502:physiology*; D016133:Polymerase Chain Reaction; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000378:metabolism*; D013601:T-Lymphocytes / Q000187:drug effects / Q000502:physiology; D013961:Thyroid Gland / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D014158:Transcription, Genetic
D015703:Antigens, CD; D000951:Antigens, Neoplasm; D000074301:CD52 Antigen; C000613862:CD52 protein, human; D006023:Glycoproteins; D011948:Receptors, Antigen, T-Cell