Detection of thyroid stimulating (TSAB)‐ and thyrotropin stimulation blocking (TSBAB) antibodies with CHO cell lines expressing different TSH‐receptor numbers

Abstract

Improvement of CHO bioassay conditions for the determination of TSH receptor antibodies leads to increased bioassay sensitivity which is most pronounced for the detection of TSBAB. Whether this increased sensitivity for the determination of TSBAB is of prognostic significance for detecting possible sequential changes in the individual TSHR AB profile of patients with Graves' disease, remains to be determined by further follow up studies.

Comparação entre diferentes métodos para avaliar a presença de auto-anticorpos dirigidos ao receptor de TSH em pacientes com moléstia de Graves-Basedow

“…No entanto, o uso de células tireóideas em cultura, quer provenientes de glândulas obtidas em cirurgia, quer em sistema celular permanente (FRTL-5), é trabalhoso e sujeito a contaminação, com resultados discordantes e contraditórios. Recentemente passou-se a usar células CHO que expressam o TSHR clonado (3,(7)(8)(9)(10). Muito recentemente outro sistema celular foi introduzido por Costagliola e cols.…”

“…De forma genérica o teste de CHO-rhTSHR/AMPc é mais sensível e específico do que o TRAb (7,8,10) atingindo cerca de 94% de positividade em pacientes com DGB não tratada. Por outro lado, o teste de CHO-rhTSH/AMPc declina, temporalmente, com o progredir da terapêutica clínica, atingindo positividade em apenas 36% dos pacientes eutireóideos após 12 meses de droga antitireóidea (7).…”

99mTc-Labeled-rhTSH Analogue (TR1401) for Imaging Poorly Differentiated Metastatic Thyroid Cancer

“…Chinese hamster ovary (CHO) JP09 and JP02 cell lines (25,26) were cultured in complete Ham's F12 medium (Eurobio Laboratories) with 10% fetal calf serum (FCS; Gibco), 100 IU/mL penicillin, 100 lg/mL streptomycin, and 2 mM l-glutamine at 37°C in a 5% CO 2 atmosphere. JP09 cells were transfected with the cDNA encoding the human TSHR and were reported to express up to 100,000 receptors per cell (25,26).…”

“…JP09 cells were transfected with the cDNA encoding the human TSHR and were reported to express up to 100,000 receptors per cell (25,26). JP02 transfected with an empty vector were used for negative controls (25,26).…”

“…JP09 cells were transfected with the cDNA encoding the human TSHR and were reported to express up to 100,000 receptors per cell (25,26). JP02 transfected with an empty vector were used for negative controls (25,26). Human ML-1 cells (27), deriving from differentiated thyroid cancer, were cultured in complete Dulbecco's modified Eagle's medium (DMEM) containing 100 lM sodium pyruvate, 2 mM l-glutamine, 1 mg/mL glucose, and 3.7 g/L NaHCO 3 , supplemented with 10% FCS, 100 IU/mL penicillin, and 100 lg/mL streptomycin.…”