A comparison of two protocols for optimal red blood cell depletion using Sepax-2 device for ABO-major incompatible transplantation in adults.

PURPOSE OF THE STUDY: In ABO-incompatible bone marrow transplantation, an efficient depletion of red blood cells (RBC) within the graft is mandatory to avoid adverse events in transplanted patients. Using non therapeutic products, we evaluated the substitution of the standard density gradient-based separation (DGBS) over Ficoll-Paque with the use of an automated procedure intended for buffy coat only (SmartRedux software) introducing modifications within the settings to achieve a drastic reduction of the initial volume of the product. Both methods were conducted on the Sepax-2 device. SAMPLES AND METHODS: RBC depletion rates and CD34+ cells recoveries from eight procedures with SmartRedux software using "in-house" settings (method A) were compared to those obtained from four procedures using NeatCell software, an automated DGBS over Ficoll-Paque (method B). RESULTS: Median erythrocyte depletion of 95,4% (92,7%-99,0%) and 99,8% (99,0%-99,9%) were observed using methods A and B, respectively. Median residual RBC volumes in the final product were 19 mL (4,4 mL-31,2 mL) and 0,7 mL (0,4 mL-4,7 mL), respectively (p = 0,014). CD34+ cells recoveries of 90,9% (62,7%-102,1%) and 78,4% (64,1%-86,2%) were achieved for methods A and B. Median platelet depletion was 16,6% (10%-42,7%) and 89,8% (88,5%-92,4%) using methods A and B, respectively (p = 0,004). Processing duration was shorter using method A (168 ± 29 min) than method B (295 ± 21 min) (p = 0,004). CONCLUSION: Both methods achieved satisfactory erythrocyte depletion and CD34+ recovery. The use of Sepax-2 device in association with SmartRedux software could be extended to efficiently deplete RBC from large-volume BM in a raw instead of DGBS.

Epstein-Barr Virus-Infected Plasma Cells Infiltrate Erosive Oral Lichen Planus.

Epstein-Barr virus (EBV), in addition to its transforming properties, contributes to the pathogenesis of several inflammatory diseases. Here, we investigated its involvement in oral lichen planus (OLP), a common autoimmune-like disease of unknown etiopathogenesis that can display a malignant potential. EBV-infected cells (EBV+ cells) were sought in a large series of clinically representative OLPs ( n = 99) through in situ hybridization to detect small noncoding EBV-encoded RNAs. Overall, our results demonstrated that EBV was commonly found in OLP (74%), with significantly higher frequency (83%) in the erosive form than in the reticular/keratinized type mild form (58%). Strikingly, many erosive OLPs were massively infiltrated by large numbers of EBV+ cells, which could represent a large part of the inflammatory infiltrate. Moreover, the number of EBV+ cells in each OLP section significantly correlated with local inflammatory parameters (OLP activity, infiltrate depth, infiltrate density), suggesting a direct relationship between EBV infection and inflammatory status. Finally, we characterized the nature of the infiltrated EBV+ cells by performing detailed immunohistochemistry profiles ( n = 21). Surprisingly, nearly all EBV+ cells detected in OLP lesions were CD138+ plasma cells (PCs) and more rarely CD20+ B cells. The presence of EBV+ PCs in erosive OLP was associated with profound changes in cytokine expression profile; notably, the expression of key inflammatory factors, such as IL1-ß and IL8, were specifically increased in OLP heavily infiltrated with EBV+ PCs. Moreover, electron microscopy-based experiments showed that EBV+ PCs actively produced EBV viral particles, suggesting possible amplification of EBV infection within the lesion. Our study thus brings conclusive evidence showing that OLP is commonly infiltrated with EBV+ PCs, adding a further puzzling element to OLP pathogenesis, given that PCs are now considered to be major regulatory immune cells involved in several autoimmune diseases (ClinicalTrials.gov NCT02276573).