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- Tollemar, Victor, et al.
(författare)
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Histopathological Grading of Oral Mucosal Chronic Graft-versus-Host Disease : Large Cohort Analysis
- 2020
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Ingår i: Biology of blood and marrow transplantation. - : Elsevier. - 1083-8791 .- 1523-6536. ; 26:10, s. 1971-1979
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Tidskriftsartikel (refereegranskat)abstract
- Graft-versus-host disease (GVHD) can manifest as acute or chronic complications in patients after hematopoietic cell transplantation (HCT). Oral chronic GVHD (cGVHD) occurs in approximately 70% of HCT recipients and includes lichenoid-like mucosal reactions, restricted mouth opening, and salivary gland dysfunction. However, the underlying histopathological presentation remains to be validated in large cohorts. We characterized the histopathological features of oral mucosal cGVHD and devised a scoring model in a large patient cohort (n = 112). Oral mucosal biopsy sections (n = 303) with and without oral cGVHD were identified from archived and current HCT recipients with additional healthy controls. Histological screening was performed on hematoxylin and eosin-stained and periodic acid-Schiff-stained sections. A points-based grading tool (0 to 19, grade 0 to IV) was established based on intraepithelial lymphocytes and band-like inflammatory infiltrate, atrophic epithelium with basal cell liquefaction degeneration, including apoptosis, as well as separation of epithelium and pseudo-rete ridges. Validation involved 62 biopsy specimens, including post-HCT (n = 47) and healthy (n = 15) specimens. Remaining biopsy specimens (n = 199) were blindly graded by 3 observers. Histological severity was correlated with clinical diagnostic and distinctive features, demonstrating a spectrum of individual patient severity, including frequent signs of subclinical GVHD in healthy mucosa. However, oral cGVHD presented with significantly higher (P < .001) scores compared with HCT controls, with moderate to high positive likelihood ratios for inflammatory infiltrate, exocytosis, and basal membrane alterations. The grade II-IV biopsy specimens demonstrated a histopathological diagnosis of active mucosal lichenoid-like cGVHD, highlighting the importance of correlating clinical presentation with the dynamic histopathological processes for improved patient stratification. In addition, this tool could be used for assessing treatments, pathological processes, and immune cellular content to provide further insight into this debilitating disease.
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| 2. |
- Tollemar, Victor, et al.
(författare)
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Immunohistopathology of oral mucosal chronic graft-versus-host disease severity and duration
- 2023
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Ingår i: Oral Diseases. - : John Wiley & Sons. - 1354-523X .- 1601-0825. ; 29:8, s. 3346-3359
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Tidskriftsartikel (refereegranskat)abstract
- Objective Chronic graft-versus-host disease (cGVHD) is the main cause of late non-relapse mortality following hematopoietic cell transplantation. Oral mucosal (om-) cGVHD is common, but diagnosis and assessment rely on clinical interpretation and patient-reported symptoms. We investigated immunohistopathological profiles with respect to om-cGVHD severity disease duration. Material and methods Ninety-four transplant patients and 15 healthy controls (n = 212 biopsies) were investigated by quantitative immunohistochemistry for T cells (CD4, CD8, and CD5), B cells (CD19 and CD20), macrophages (CD68), and Langerhans cells (CD1a). Results We found significant increases in T (CD4, CD8) and monocytic (CD68) cells in om-cGVHD, and a notable absence of B (CD19 and CD20) cells. Histopathological activity correlated with increased CD4, CD8 and CD68. However, CD4 was associated with mild om-cGVHD, whereas CD8 and CD68 were found to be elevated in severe om-cGVHD. CD8 and CD68 levels were raised at disease onset, but during late phase, the predominant CD68 population was accompanied by CD4. Conclusion Oral cGVHD is a heterogenous clinical disorder, but our knowledge of the underlying biology remains limited. We highlight the importance of CD4, CD8 and CD68 immune profiling, together with histological grading for the staging of oral cGVHD, to broaden our understanding of the biology and individual disease course.
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| 3. |
- Tollemar, Victor
(författare)
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Oral chronic Graft-versus-host disease : clinical and pathological staging
- 2022
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Allogenic hematopoietic cell transplantation (HCT) is a curative treatment for many patients with immune- hematopoietic disorders, mainly hematopoietic cancers as leukaemia. Chronic Graft-versus-Host Disease (cGVHD) is a major long-term complication, associated with mortality and morbidity following allogenic HCT. Oral cGVHD is common and might manifest as mucosal lichenoid manifestations (om-cGVHD) or with dysfunctional salivary glands (sg-cGVHD). Alloreactive T-cells respond to recipient tissues with pathological reactions of acute inflammation, progressing with chronic inflammation and dysregulated immunity, and subsequent aberrant fibrotic healing. This thesis aimed to investigate diagnostic criteria for oral cGVHD using histopathological, clinical, and immune cell characterisation. A retrospective cohort of 95 HCT-patients and 303 oral biopsies were analysed, including 15 healthy controls. Oral mucosal biopsies with and without minor salivary glands (MSG) were retrieved from Stockholm Medical Biobank. Associated clinical information was gathered from the clinical charts and HCT register data. We applied histological (Haematoxylin and Eosin, Periodic acid Schiff, van Gieson), and immunohistochemical (IHC) staining (CD4 T-helper cells, CD8 T-cytotoxic cells, CD68 macrophages, CD1a Langerhans cells (LCs), CD19 and CD20 B-cells, and CD5 T-/B-cells). Quantitative IHC was performed using CellProfiler image analysis software. In papers I and IV, oral mucosal-, and MSG histopathology were analysed in biopsies prior and post HCT, with and without cGVHD. We used the National Institutes of Health pathology criteria and formalised grading modules to assess pathology scores and grades (NIH cGVHD grading). The oral mucosa was observed with minimal criteria of lichenoid interface inflammation with exocytosis, liquefaction degeneration and apoptosis. Basal membrane alterations were the most specific criteria found. Features detected in the MSG were peri-ductal and acinar inflammation and exocytosis, destruction and fibrosis. We developed severity grades (G)0-IV and verified pathology diagnostics of “possible (GII)” and “likely (GIII-GIV)” cGVHD. In paper IV, we also employed the Greenspan composite MSG grading scheme, which was found with a strong correlation to the NIH cGVHD MSG grading. IHC quantification was performed following established pipelines in CellProfiler, as described in paper II. The methodology was compared to manual counting, with a perfect concordance in detection of positive stained cells, as well as for positive stained regions. The benefit of CellProfiler is to perform standardised and repeatable quantification in a time saving manner. Oral mucosal immune profiles were investigated in paper III. CD4 infiltration was associated with mild and distinctive om-cGVHD but were found with frequent stable levels over time. CD8 was elevated in clinical and pathologically severe om-cGVHD, particularly during cGVHD onset and progression. Immunolocalisation of CD68 revealed significant staining in various clinical groups, particularly at onset, but the association with severity was interesting especially during late stages of disease. CD1a LCs were significantly reduced in pathological GII at onset and during progression, but otherwise non-significant compared to healthy. CD19 and CD20 were rarely observed. In paper IV, we quantified the immune profiles in the MSG and found an altered pathology with significant increase of CD4- and CD8-cells. However, levels of B-cells and LCs were considerably low. The association between oral mucosal and MSG immunopathology, was investigated on the whole cohort and with respect to cGVHD duration. Overall, a moderate correlation was observed for pathology scores, CD4 and CD8 infiltrate. Interestingly, at the time of cGVHD onset, the correlation between the oral mucosal disease and MSG was stronger but with progression no further association was found. In conclusion, om- and sg-cGVHD are two heterogenous complications that display associated immune-pathology profiles during cGVHD onset, but progression appears to be tissue- dependent. We developed histopathological grading modules to facilitate severity diagnostics, which were significantly associated with CD4, CD8 and CD68 immunostaining. om-cGVHD clinical and pathological characterisation was found associated to changes in the immune profile. CD8 was found to drive the severe disease reaction during onset and progression but diminished over time. However, long duration of the disease correlated with elevated CD68 and persistent CD4 cells. This highlights the need for improved clinical and pathological characterisation in combination with biological disease classification.
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