Common Histological Features Suggesting Enchondral Ossification Pathways in Calciphylaxis of Various Origins: A Study of Human Subcutaneous Tissue Biopsies.

Calciphylaxis is a rare, yet underdiagnosed condition causing high mortality in patients with severe renal and cardiovascular disease. Since knowledge of the pathophysiology of calciphylaxis is limited, a differential analysis of histological alterations in patient subgroups with various comorbidities might expose different disease phenotypes and allow deeper insights into the pathophysiology of the condition. Histological markers of osteogenesis and calcification were investigated in a group of 18 patients with clinically and histologically verified calciphylaxis, using immunohistochemical staining. Analysis of staining intensity and distribution of marker proteins in histological structures was performed to evaluate distinct patterns between subgroups with different clinical comorbidities in comparison with a control group. In all cases, immunohistochemical staining for bone matrix proteins, bone-morphogenic proteins and matrix-Gla proteins co-localized with subcutaneous vascular and interstitial calcifications. Significant expression of bone-morphogenic protein-7 and active matrix-Gla protein was observed. Mortality was associated with renal comorbidities and increased expression of bone-morphogenic protein-7. However, no distinct histological patterns were found between subgroups with renal disease, warfarin intake or coexisting micro- and macro-angiopathies. The upregulation of osteogenic markers (including bone-morphogenic protein-7) plays a major role in the development of calciphylaxis. Clinical outcome correlates with kidney function and phosphate handling, suggesting different pathophysiological mechanisms. However, biopsy  at late-stage disease shows a common histological phenotype, involving enchondral ossification.

Leukocyte-Reactive Antibodies in Female Blood Donors: The Austrian Experience.

INTRODUCTION: Antibody-mediated transfusion-related acute lung injury (TRALI) is caused by antibodies against human leukocyte antigens (HLAs) or human neutrophil antigens (HNAs), and is one of the most serious complications associated with transfusion medicine. Prevention strategies like testing allo-exposed female blood donors have not yet been introduced nationwide in Austria. To assess the need and feasibility of routine leukocyte antibody testing, the prevalence of leukocyte-reactive antibodies in an Austrian female donor population was been determined using classical cell-based methods which were compared with a high-throughput bead-based method. METHODS: Sera from 1,022 female blood donors were screened using a granulocyte aggregation test (GAT) and a white blood cell immunofluorescence test (WIFT) after retesting and specification of positive samples by granulocyte immunofluorescence test (GIFT) and monoclonal antibody-specific immobilization of granulocyte antigens (MAIGA). Potential HLA reactivities were confirmed using the microbeads assay LabScreenTM Mixed. The results in 142 donor sera and 38 well-defined reference sera were investigated by the microbeads assay LabScreenTM Multi and compared with classical cell-based methods. RESULTS: Reactivity with either granulocytes and/or lymphocytes was detected in 79 sera (7.7%), with the majority being HLA-specific. Antibodies against HNA were obtained in 7 samples (0.7%). The aggregating potential of the detected antibodies was observed in 9 cases (0.9%). Most of the leukocyte-reactive antibodies occurred at a donor age of between 35 and 59 years (n = 61). LabScreen Multi showed good agreement (κ = 0.767) for HNA antibody detection by cell-based assays, but double/multiple specificities (100% of 7 anti-HNA-1b sera) as well as false-negative results (40% of 15 HNA-3-specific sera) occurred. DISCUSSION: Leukocyte-reactive antibody screening is advised in Austrian female donors for safe blood transfusion, including single-donor convalescent plasma treatment of COVID-19 that may be implemented soon. For the introduction of LabScreen Multi, the combination with GAT should be considered to ensure correct anti-HNA-3a detection.