Increased circulating Tfh17 and PD-1+Tfh cells are associated with autoantibodies in Hashimoto's thyroiditis.

OBJECTIVE: Hashimoto's thyroiditis (HT) is characterized by autoantibodies targeting the thyroid. Abnormal CD4+CXCR5+T cell levels were previously shown to be associated with HT. However, Tfh cells consist of heterogeneous subpopulations, and which T follicular helper (Tfh) cell subpopulation participates in the pathogenesis of HT remains poorly understood. METHODS: Thirty healthy controls (HCs) and 52 HT patients were enrolled in the study. The percentages of Tfh, ICOS+Tfh, PD1+Tfh, Tfh1, Tfh2, Tfh17, effector Tfh, resting Tfh, effector memory Tfh, central memory Tfh, and naïve Tfh cells in the peripheral blood were all determined via flow cytometry, and the associations between the percentages of these cells and thyroid function indices were also investigated. RESULTS: The percentage of Tfh cells was significantly higher in HT patients than in HCs. Examination of the Tfh cell subsets revealed that the percentages of Tfh1, Tfh2, and resting Tfh cells were significantly decreased, while those of the ICOS+Tfh, PD1+Tfh, Tfh17, and effector Tfh cells were significantly increased in HT patients. No significant differences in effector memory, central memory or naïve Tfh cell percentages were noted between the HC and HT groups. Furthermore, the percentage of PD1+Tfh cells was positively correlated with anti-thyroglobulin antibody levels. Most importantly, only Tfh17 cell percentages were positively correlated with anti-thyroglobulin and anti-thyroid peroxidase antibody levels and were negatively correlated serum free T3 and free T4 levels in HT patients. CONCLUSIONS: Increased circulating Tfh17 cell and PD1+Tfh percentages are associated with higher autoantibody levels in HT patients, which imply that Tfh17 or PD1+Tfh cells may play a pathogenic role in the development of HT.

A New Method of Using Polyethylene Glycol (PEG) Precipitation of Macroprolactin to Detect Genuine Hyperprolactinemia.

BACKGROUND: The most commonly used method of polyethylene glycol (PEG) precipitation for macroprolactinemia (MP) screening has some significant drawbacks. The aim of this study was to establish a new method using PEG for precipitation of macroprolactin (macroPRL) to detect genuine hyperprolactinemia (genuine HP). METHODS: The optimal PEG concentration for precipitation and the effect of PEG on the precipitation of PRL were analyzed to establish and optimize our PEG precipitation method. The PRL recovery rate and genuine HP detection rate were compared between our method and MP screening method. RESULTS: About 25% PEG6000 was determined to be the optimal PEG concentration for precipitation. Along with an increase in protein concentration in the PRL calibration solution, the PRL recovery rate after precipitation decreased gradually. The PRL recovery rate increased when the precipitation was carried out with diluted PRL calibration solution; the recovery rate reached greater than 90% after a 5-fold dilution of the calibration solution. The genuine HP detection rate and PRL recovery rate using our diluted serum PEG precipitation method were significantly higher than those obtained with the MP screening method. Our method successfully detected 31 cases of genuine HP, which was significantly higher than the detection rate obtained using the MP screening method (25 cases; P < 0.001). CONCLUSION: Precipitation using 5-fold diluted serum with 25% PEG6000 can effectively reduce the macroPRL concentration, increasing the PRL recovery rate and detection rate of genuine HP after precipitation, which is an effective and convenient method for the detection of genuine HP.