Expansion of effector memory Vδ2neg γδ T cells associates with cytomegalovirus reactivation in allogeneic stem cell transplant recipients.

Background: Cytomegalovirus (CMV) reactivation is a significant concern following allogeneic stem cell transplantation. While previous research has highlighted the anti-CMV reactivation effect of γδ T cells in immunocompromised transplant patients, their characterization in recipients at high risk of CMV reactivation remains limited. Methods: This study focused on D+/R+ recipients (where both donor and recipient are CMV seropositive) at high risk of CMV reactivation. We analyzed 28 patients who experienced CMV recurrence within 100 days post-allogeneic hematopoietic stem cell transplantation, along with 36 matched recipients who did not experience CMV recurrence. Clinical data from both groups were compared, and risk factors for CMV reactivation were identified. Additionally, CMV viral load was measured, and flow cytometric analysis was conducted to assess changes in peripheral blood γδ T cell proportions, subpopulation distribution, and differentiation status. We also analyzed the CDR3 repertoire of the TCR δ chain in different γδ T cell subsets. Functional analysis was performed by measuring the lysis of CMV-infected cells upon stimulation. Results: CMV reactivation post-transplantation was associated with acute graft-versus-host disease (aGvHD) and reactivation of non-CMV herpesviruses. Notably, CMV reactivation led to sustained expansion of γδ T cells, primarily within the Vδ2neg γδ T cell subpopulation, with a trend toward differentiation from Naive to effector memory cells. Analysis of the δ chain CDR3 repertoire revealed a delay in the reconstitution of clonal diversity in Vδ2neg γδ T cells following CMV reactivation, while Vδ2pos T cells remained unaffected. Upon stimulation with CMV-infected MRC5 cells, the Vδ2neg γδ T cell subpopulation emerged as the primary effector cell group producing IFN-γ and capable of lysing CMV-infected cells. Moreover, our findings suggest that NKG2D is not necessary involved in Vδ2neg γδ T cell-mediated anti-CMV cytotoxicity. Conclusion: This study provides novel insights into the role of γδ T cells in the immune response to CMV reactivation in transplantation recipients at high risk of CMV infection. Specifically, the Vδ2neg γδ T cell subpopulation appears to be closely associated with CMV reactivation, underscoring their potential role in controlling infection and reflecting CMV reactivation in HSCT patients.

Impact of Corticosteroids on the Proportions of Circulating Tfh Cell Subsets in Patients With Systemic Lupus Erythematous.

Objectives: This study aimed to analyze the distribution of T follicular helper (Tfh) cells in lupus patients, and the effects of steroids on circulating Tfh cells. Methods: Circulating Tfh cell subsets were defined by multicolor flow cytometry as Tfh17, Tfh2 or Tfh1 subpopulations of CXCR5+CD45RA-CD4+ T cells in the peripheral blood of SLE patients and healthy controls. To test the effects of corticosteroid on Tfh cells, PBMC harvested from both SLE and healthy controls were cocultured with dexamethasone, and then analyzed by Flow cytometry. Results: The proportion of Tfh17 cells in SLE patients was increased significantly compared with healthy controls. Additionally, patients with an active disease had reduced Tfh1 subsets than those with an inactive disease and healthy controls. The frequency of Tfh2 cells was associated with the proportion of circulating plasmablasts and the amount of anti-dsDNA. Dexamethasone reduced the percentage of Tfh2 cells while increased the proportion of Tfh17 subset in gated CXCR5+CD45RA-CD4+ T cells. Conclusion: Our study investigated the distribution of circulating Tfh subsets in lupus patients. Corticosteroids treatment not only down-regulated the proportion of circulating Tfh cells, but also altered the distribution of Tfh subsets in vivo and in vitro.

Performance evaluation of the new platelet measurement channel on the BC-6800 Plus automated hematology analyzer.

OBJECTIVE: To evaluate the performance of new optical platelet measurement channel on the BC-6800 Plus automated blood cell analyzer. METHODS: The basic PLT count performance of the BC-6800 Plus was evaluated according to the requirements of the Clinical Laboratory and Standards Institute (CLSI) Document H26-A2. In addition, low-PLT-value specimens, red blood cell debris specimens, small red blood cell specimens, and giant PLT specimens were detected with the blood cell analyzer and a flow cytometer. Whole-blood specimens in ethylenediaminetetraacetic acid (EDTA) or sodium citrate anticoagulant tubes from 20 patients with EDTA-dependent PLT aggregation were determined in CDR mode of the analyzer. RESULTS: Blank counting and the carryover contamination rate of PLTs using the BC-6800 Plus both met the technical requirements. For abnormal PLT specimens, PLT-O 8× and PLT-I both exhibited high comparability with flow cytometry. The comparability of PLT-O 8× with flow cytometry was better than that of PLT-I. In EDTA-anticoagulated blood specimens from 20 patients with EDTA-dependent PLT aggregation, the results of PLT-O were significantly higher than those for PLT-I using samples from the same tubes (P < .001). However, the PLT counts were similar between these two methods for sodium citrate-anticoagulated blood specimens (P = .263). CONCLUSION: The performance of PLT-O 8× in the BC-6800 Plus met the technical requirements. PLT-O 8× exhibited better reproducibility than did PLT-I for low-PLT-value samples. Reexamination of abnormal PLT specimens using PLT-O 8× yielded more accurate results. PLT-O performed significantly better than PLT-I in the detection of EDTA-dependent PLT-aggregation specimens.

Comparison of Ultrasound-Guided Fine-Needle Cytology Quality in Thyroid Nodules with 22-, 23-, and 25-Gauge Needles.

OBJECTIVE: To compare the cytology quality of ultrasound-guided fine-needle biopsy in thyroid nodules with 22-, 23-, and 25-gauge (G) needles prospectively. METHODS: A total of 240 consecutive nodules underwent ultrasound-guided fine-needle aspiration (USG-FNA) and 240 nodules underwent ultrasound-guided fine-needle capillary (USG-FNC) were included in this prospective study from October 2014 to February 2016. Each nodule was sampled using 22 G, 23 G, and 25 G needle according to designed orders, and 1240 smears were finally obtained. Cytology quality was scored by a cytologist blinded to needle selection. RESULTS: In USG-FNA, the average scores and standard deviations were 5.50 ± 2.87 for 25 G needles, 4.82 ± 2.95 for 23 G needles, and 5.19 ± 2.81 for 22 G needles. In USG-FNC, the average scores and standard deviations of each group were 5.12 ± 2.69 for 25 G, 4.60 ± 2.90 for 23 G, and 4.90 ± 2.90 for 22 G needles. The specimen quality scores of 25 G group were significantly higher than that of 23 G group (P < 0.017) in both USG-FNA and USG-FNC. However, the differences were not statistically significant in nondiagnostic rate using different gauge of needles (P > 0.017 for all). CONCLUSIONS: 25 G needles obtained the highest scores of sample quality in thyroid FNA and FNC comparing with 22 G and 23 G needles. 25 G needle should be first choice of thyroid FNA and FNC in routine work.

The thromboelastography G parameter as a potential biomarker of acute coronary syndrome.

The most prominent event that defines acute coronary syndrome (ACS) is the formation of an intra-arterial thrombus, usually resulting from activation of platelet and fibrinogen at the ruptured plaque. Usually, conventional coagulation tests (CCTs) are used to estimate the hemostatic properties of patients. However, CCTs have significant limitations because they each assess individual aspects of the coagulation cascade, which is a complex multifaceted process. And CCTs are performed with platelet-poor plasma, while the contribution of platelets to clot formation is not measured. In contrast, thromboelastography (TEG) is a test for global hemostasis with whole blood, from the beginning of coagulation through clot formation to the ending with fibrinolysis. The aim of this study was to investigate whether TEG parameters could be surrogate biomarkers of thrombus formation process and diagnosis of ACS. Receiver operating characteristic（ROC）curve was used to evaluate the diagnosis performance of each index. Logistic regression analysis was utilized to define the independent risk factors of ACS. The results showed that the shear elastic modulus parameter (G) was an independent diagnostic indicator for ACS (odds ratio [OR], 2.600; 95% confidence interval [CI], 2.035-3.322). The area under ROC curve of G was 0.866. The optimal cut-off value for the diagnosis of ACS was 10.55 dyne/cm2, while the sensitivity was 66.2% and the specificity was 92.4%. In conclusion, G could be used as an optimal indicator of activation of platelet and fibrinogen, which is eligible to be a useful biomarker for early diagnosis of ACS.

Size and Ultrasound Features Affecting Results of Ultrasound-Guided Fine-Needle Aspiration of Thyroid Nodules.

OBJECTIVES: Our goal was to assess the diagnostic efficacy of ultrasound (US)-guided fine-needle aspiration (FNA) of thyroid nodules according to size and US features. METHODS: A retrospective correlation was made with 1745 whole thyroidectomy and hemithyroidectomy specimens with preoperative US-guided FNA results. All cases were divided into 5 groups according to nodule size (≤5, 5.1-10, 10.1-15, 15.1-20, and >20 mm). For target nodules, static images and cine clips of conventional US and color Doppler were obtained. Ultrasound images were reviewed and evaluated by two radiologists with at least 5 years US working experience without knowing the results of pathology, and then agreement was achieved. RESULTS: The Bethesda category I rate was higher in nodules larger than 15 mm (P < .05). The diagnostic accuracy was best in nodules of 5 to 10 mm in diameter. The sensitivity, accuracy, PPV, and LR for negative US-guided FNA results were better in nodules with a size range of 5 to 15 mm. The specificity, negative predictive value (NPV), and LR for positive results and the Youden index rose with increasing nodule size. Seventeen false-positive and 60 false-negative results were found in this study. The false-negative rate rose with increasing nodule size. However, the false-positive rate was highest in the group containing the smallest nodules. Nodules with circumscribed margins and those that were nonsolid and nonhypoechoic and had no microcalcifications correlated with Bethesda I FNA results. Nodules with circumscribed margins and those that were nonsolid, heterogeneous, and nonhypoechoic and had increased vascularity correlated with false-negative FNA results. Borders correlated with Bethesda I false-negative and false-positive FNA results. CONCLUSIONS: Tiny nodules (≤5 mm) with obscure borders tended to yield false-positive FNA results. Large nodules (>20 mm) with several US features tended to yield false-negative FNA results.