METHODS: Two tissue cohorts containing 98 HHs and 13 HEHEs were used to study entity-specific and disease stage-specific endothelial cell (EC) phenotype and immune cell abundance. Using semiquantitative assessment, annotation-based cell classifiers, digital cell detection on whole slides, and tissue microarrays, we quantified 23 immunologic and vascular niche-associated markers and correlated this with clinicopathologic data.
RESULTS: Both HH and HEHE ECs were characterized by a CD31high, CD34high, FVIII-related antigenhigh expression phenotype with entity-specific expression differences of sinusoidal EC markers Stabilin1, Stabilin2, CD32, and Lymphatic Vessel Endothelial Hyaluronan Receptor 1 (LYVE-1). Cell detection identified an HH margin-prevailing immunologic response dominated by Myeloperoxidase+ (MPO+) macrophages, CD3+ and CD8+ T cell subsets, and B cells (CD20+, CD79A+). In HEHE, increased CD68+ and CD20+ cell demarcation of lesion margins was observed, while CD3+ and CD8+ T cells were equally detectable both marginally and intralesionally. Stage-specific pairwise correlation analysis of HH and HEHE revealed disease entity-specific immunologic infiltration patterns as seen by high CD117+ cell numbers in HH, while HEHE samples showed increased CD3+ T cell infiltration.
CONCLUSIONS: ECs in HH and HEHE share a continuous EC expression phenotype, while the expression of sinusoidal EC markers is more highly retained in HEHE. These phenotypic differences are associated with a unique and disease-specific immunovascular landscape.
方法:使用包含98个HHs和13个HEHE的两个组织队列来研究实体特异性和疾病阶段特异性内皮细胞(EC)表型和免疫细胞丰度。使用半定量评估,基于注释的单元格分类器,整个幻灯片上的数字细胞检测,和组织微阵列,我们量化了23项免疫和血管生态位相关标志物,并将其与临床病理数据相关联.
结果:HH和HEHEECs均以CD31高,CD34高,FVIII相关抗原高表达表型与正弦EC标记物Stabilin1、Stabilin2、CD32和淋巴管内皮透明质酸受体1(LYVE-1)的实体特异性表达差异。细胞检测确定由髓过氧化物酶+(MPO+)巨噬细胞主导的HH边缘占优势的免疫应答,CD3+和CD8+T细胞亚群,和B细胞(CD20+,CD79A+)。在赫赫,病变边缘CD68+和CD20+细胞分界增加,而CD3+和CD8+T细胞在边缘和细胞内同样可检测到。HH和HEHE的阶段特异性成对相关性分析揭示了疾病实体特异性免疫浸润模式,如HH中的高CD117细胞数所示,而HEHE样本显示CD3+T细胞浸润增加。
结论:HH和HEHE中的EC具有连续的EC表达表型,而正弦EC标志物的表达在HEHE中保留得更高。这些表型差异与独特的疾病特异性免疫血管景观有关。