Significance of CD10 protein expression in the diagnostics of follicular lymphoma: A comparison of conventional immunohistochemistry with flow cytometry supported by the establishment of BCL2 and BCL6 rearrangements.

INTRODUCTION: Histopathological examination and immunohistochemistry (IHC) with a crucial role of CD10 expression remain a standard diagnostic tool in follicular lymphoma (FL). The results of IHC CD10 detection with different primary antibodies are not fully reproducible, but some reports show that flow cytometry (FCM) can be a reliable method of CD10 identification. METHODS: The aim of the study was to compare results of CD10 expression in FL by IHC and FCM including immunophenotypic features in the context of the BCL2 and BCL6 alterations. RESULTS: Out of 76 histopathologically diagnosed FL, a group of 25 cases had simultaneously FCM. Immunohistochemically 77.6% of cases were CD10-positive with comparable and reproducible results to FCM. Differences between the FCM expression of CD5/CD10/CD11c/CD25/CD43 and BCL2 overexpression (BCL2(+)higher ) correlated with the BCL2 and BCL6 rearrangements (R) status. Lack of CD10 expression corresponded with the absence of BCL2R and higher MUM1 expression by IHC results but had no clinical impact on the long-time outcomes. CONCLUSIONS: Immunohistochemistry staining is a comparable method to FCM assessment in the evaluation of CD10 expression and diagnosis of FL. Fine-needle aspiration biopsy/FCM (FNAB/FCM) could be a useful tool for verifying FL diagnosis and CD10 detection. Despite its heterogeneity, FL has a characteristic immunophenotype. BCL2R and BCL6R FL cases differ mainly in levels of BCL2 and CD10 with CD43 co-expression; BCL2(+)higher by FCM correlates with BCL2R. Moreover, FNAB plays an important role in material provision for supportive karyotyping and BCL2R, BCL6R assessed by FISH.

The distinct clinical features and prognosis of the CD10⁺MUM1⁺ and CD10⁻Bcl6⁻MUM1⁻ diffuse large B-cell lymphoma.

Using an immunohistochemistry (IHC) based method, diffuse large B-cell lymphoma (DLBCL) can be classified into germinal center B-cell (GCB) and non-GCB subtypes. However, the prognostic value of Hans algorithm was contradictory in the literature. Using IHC and fluorescence in situ hybridization, we analyzed the antibodies applied in Hans algorithm and other genetic factors in 601 DLBCL patients and prognostic value of Hans algorithm in 306 cases who were treated with chemoimmunotherapy. The results showed that patients with GCB subtype have better overall survival (OS) and progression-free survival (PFS) than non-GCB cases. However, to some extent, double positive (CD10(+)MUM1(+), DP) and triple negative (CD10(-)Bcl6(-)MUM(-), TN) showed different clinical characteristics and prognosis to others that were assigned to the same cell-of-origin group. The DP group showed similar OS (median OS: both not reached, P = 0.3650) and PFS (median PFS: 47.0 vs. 32.7 months, P = 0.0878) with the non-GCB group while the TN group showed similar OS (median OS: both not reached, P = 0.9278) and PFS (median PFS: both not reached, P = 0.9420) with the GCB group. In conclusion, Recognition of specific entities in Hans algorithm could help us to accurately predict outcome of the patients and choose the best clinical management for them.

MiR-346 regulates CD4⁺CXCR5⁺ T cells in the pathogenesis of Graves' disease.

Follicular helper T (Tfh) cells are increasingly recognized as participants in various autoimmune diseases, including Graves' disease. Although many transcription factors and cytokines are known to regulate Tfh cells, the role of noncoding RNA in Tfh cells development and function is poorly understood. Twenty-three patients with GD, eleven patients with remitting GD, and twenty-four healthy controls were enrolled in the current study. The interaction of miRNA and target gene was predicted through software analysis and then validated by luciferase assay and Western blot. The levels of miR-346 in circulating CD4(+) T cells and plasma were measured by qRT-PCR. The correlation of miR-346 levels with the percentages of CD4(+)CXCR5(+)T cells and autoantibody levels were also analyzed. Up-regulation of Bcl-6 and down-regulation of miR-346 in GD patients were observed, and miR-346 could inhibit Bcl-6 at both transcriptional and translational levels. Overexpression of miR-346 led to attenuating CD4(+)CXCR5(+) T cells. The abnormal expression of miR-346 restored in GD patients after treatment. A negative correlation between levels of miR-346 and percentages of CD4(+)CXCR5(+) T cells was confirmed in GD patients. Additionally, negative correlations between the levels of miR-346 in circulating CD4(+) T cells and serum concentrations of TR-Ab, TG-Ab, and TPO-Ab were also revealed in GD patients. MiR-346 regulates CD4(+)CXCR5(+) T cells by targeting Bcl-6, a positive regulator of Tfh cells, and might play an important role in the pathogenesis of Graves' disease.