Immunohistopathology of oral mucosal chronic graft‐versus‐host disease severity and duration

Abstract

Post-transplant, patients treated with allogeneic hematopoietic cell transplantation (HCT) commonly develop acute(a) and/or chronic(c) graft-versus-host disease (GVHD) (Lee, 2017). cGVHD, an autoimmune-like disorder, can develop at multiple sites, and 45%-83% of patients develop oral manifestations (

Grading of minor salivary gland immuno-histopathology post-allogenic hematopoietic cell transplantation

“…The patients analyzed in this study are from a retrospective cohort of allogenic-HCT patients treated at Karolinska University Hospital (KUH) between 1977 and 2011, with biopsies retrieved from Stockholm′s Medicine Biobank (SMB) [ 26 ]. The cohort has been described previously for the studies of the om-cGVHD histopathological profile [ 26 , 27 ]. Biopsies had been routinely obtained as standard of care at the Oral and Maxillofacial Surgery Clinic (OMSC) KUH over the first year post-HCT at 3-month intervals, and thereafter on an individual basis for later time points (typically obtained at 3-, 6-, 9-, 12-, and ≥24-months), which in some cases resulted in more multiple biopsies per patient.…”

“…Slides were stained with H&E (Histolab Products AB, Gothenberg, Sweden) for routine pathology, as well as PAS (Merck KGaA, Germany) for mucopolysaccharides and van Gieson (Sigma-Aldrich, Germany) for fibrosis according to the manufacturer’s instructions (Malkusch et al, 1995). Immunohistochemistry (IHC) was performed using the monoclonal antibodies, CD1a (1:15,000), CD5 (1:300), CD8 (1:1000), CD19 (1:150), CD20 (1:400) and CD68 (1:30,000) (DAKO)) according to the protocol described in Tollemar et al, 2022 [ 27 ]. All glass slides were digitalized at 40x magnification using 3D Histech Scanner and viewed in CaseViewer (Histolab Products AB).…”

“…Quantitative-IHC used CellProfiler version 3.1.9 ( www.cellprofiler.org ) and pipelines were established for each antibody [ [27] , [28] , [29] ]. Images were segmented into 1000 × 1000 pixel tiles (>15 kb) using ImageMagick version 7.0.8 ( www.imagemagick.org ).…”

“…Images with folds and artefacts were excluded. The remaining tiles were recombined to give total stained pixel area/total pixel area, and the average determined per biopsy for each antibody [ [27] , [28] , [29] ].…”

“…As a result, we defined guided pathological diagnostics of “possible” and “likely” cGVHD [ 8 , 18 , 19 ]. In addition, MSG immune cell phenotyping with the histopathological grading was compared for disease severity [ 27 ]. We hypothesized that MSG immunopathological staging will increase our understanding of oral cGVHD pathophysiology’s [ 26 , 27 ].…”