No major monoclonal lymphocyte population in the thyroid of patients with Graves' disease: study of gene rearrangement by restriction fragment length polymorphism and polymerase chain reaction.
The present study aimed at determining the mono-, oligo-, or polyclonal nature of intrathyroid lymphocytes at the DNA level in patients with Graves' disease. Two techniques were used to seek monoclonal rearrangement in DNA derived from intrathyroidal lymphocytes obtained from six patients. The first was restriction fragment length polymorphism using two specific probes from the B-chain of T-cell receptor and the other from the heavy chain immunoglobulin gene; the second was polymerase chain reaction using a couple of specific primers from the variable and joining regions of heavy chain immunoglobulins. The results for the patients with Graves' disease were compared with those obtained for circulating T-and B-lymphocytes, granulocytes (negative controls), and T- and B-leukemic cells (positive controls). The results with restriction fragment length polymorphism favored a polyclonal origin for the lymphocytes in all cases, since no rearrangement was visualized. The results with polymerase chain reaction were analogous, and the technique was 10 times more sensitive in the detection of rearrangement.
Department of Endocrinology, Hopital Fontenoy, Rennes, France.
C
C
Petit-Simon
G
G
Semana
F
F
Quillivic
R
R
Fauchet
B
B
Genetet
H
H
Allannic
D000328:Adult; D001402:B-Lymphocytes / Q000276:immunology*; D001483:Base Sequence; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D015245:Deoxyribonuclease BamHI; D015247:Deoxyribonuclease HindIII; D005260:Female; D015321:Gene Rearrangement; D006111:Graves Disease / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D011947:Receptors, Antigen, B-Cell / Q000235:genetics*; D011948:Receptors, Antigen, T-Cell / Q000235:genetics*; D013601:T-Lymphocytes / Q000276:immunology*; D013961:Thyroid Gland / Q000276:immunology*
D009838:Oligodeoxyribonucleotides; D011947:Receptors, Antigen, B-Cell; D011948:Receptors, Antigen, T-Cell; D004247:DNA; D015245:Deoxyribonuclease BamHI; D015247:Deoxyribonuclease HindIII