3D analyses reveal T cells with activated nuclear features in T-cell/histiocyte-rich large B-cell lymphoma

Abstract

Nodular lymphocyte-predominant Hodgkin lymphoma (NLPHL) can show variable histological growth patterns and present remarkable overlap with T-cell/histiocyte-rich large B-cell lymphoma (THRLBCL). Previous studies suggest that NLPHL histological variants represent progression forms of NLPHL and THRLBCL transformation in aggressive disease. Since molecular studies of both lymphomas are limited due to the low number of tumor cells, the present study aimed to learn if a better understanding of these lymphomas is possible via detailed measurements of nuclear and cell size features in 2D and 3D sections. Whereas no significant differences were visible in 2D analyses, a slightly increased nuclear volume and a significantly enlarged cell size were noted in 3D measurements of the tumor cells of THRLBCL in comparison to typical NLPHL cases. Interestingly, not only was the size of the tumor cells increased in THRLBCL but also the nuclear volume of concomitant T cells in the reactive infiltrate when compared with typical NLPHL. Particularly CD8+ T cells had frequent contacts to tumor cells of THRLBCL. However, the nuclear volume of B cells was comparable in all cases. These results clearly demonstrate that 3D tissue analyses are superior to conventional 2D analyses of histological sections. Furthermore, the results point to a strong activation of T cells in THRLBCL, representing a cytotoxic response against the tumor cells with unclear effectiveness, resulting in enhanced swelling of the tumor cell bodies and limiting proliferative potential. Further molecular studies combining 3D tissue analyses and molecular data will help to gain profound insight into these ill-defined cellular processes.

Tumour cell characteristics and microenvironment composition correspond to clinical presentation in newly diagnosed nodular lymphocyte‐predominant Hodgkin lymphoma

“…On the one hand, larger tumour cell nuclei were observed in cases with variant growth patterns, possibly reflecting the presence of a higher number of genomic aberrations, mutations and overall genomic complexity 20,21 . This is in line with a previous study, in which a slightly larger nuclear area and volume in the LP cell nuclei were observed in NLPHL cases with growth pattern C 22 . On the other hand, patients with particularly large tumour cell nuclei had more limited disease supporting the hypothesis that larger nuclear size may hamper tumour cell dissemination.…”

“…39 It has been shown that tissue macrophages in T-cell/histiocyte-rich large B-cell lymphoma express PD-L1 40 and thus probably dampen an anti-tumour cytotoxic immune response. 22 In summary, the present study for the first time elucidates tumour cell characteristics and features of the microenvironment in NLPHL in relation to the clinical presentation, expanding our understanding of the development and progression of this lymphoma entity. We have observed a correlation between both LP cell nuclear size as well as amount and interaction of PD1-positive T FH cells with clinical stages.…”

“…Especially the microenvironment of NLPHL presenting with pattern E has a strong histological and molecular similarity to T‐cell/histiocyte‐rich large B‐cell lymphoma 39 . It has been shown that tissue macrophages in T‐cell/histiocyte‐rich large B‐cell lymphoma express PD‐L1 40 and thus probably dampen an anti‐tumour cytotoxic immune response 22 …”

Spatial Phenotyping of Nodular Lymphocyte Predominant Hodgkin Lymphoma and T-cell/Histiocyte-Rich Large B-cell Lymphoma

“…Higher cytotoxic and activated cytotoxic T-cells content in THRLBCL compared to NLPHL of all patterns, particularly pattern E, suggests a potential role for activation status of cytotoxic T-cells in orchestrating architectural dismantling in the transition from NLPHL to THRLBCL. Our data also con rm high CD8 content in THRLBCL, [17,48] and higher cytotoxic T-cell/monocyte interactions in THRLBCL compared with NLPHL, [49] although the functional status of cytotoxic T-cells in driving NLPHL transformation remains unknown. In addition, we show that variant NLPHL patterns, especially pattern E, has stronger cytotoxic T-cell interactions with LP cells compared to typical nodular patterns of NLPHL.…”

Three-dimensional Isotropic Imaging of Live Suspension Cells Enabled by Droplet Microvortices

“…38 Additionally, the study of other diseases, such as Nodular lymphocyte-predominant Hodgkin lymphoma (NLPHL) and T-cell/histiocyte-rich large B-cell lymphoma (THRLBCL), further emphasizes the importance of nuclear 3D analyses in activated T cells. 39 Particularly, whereas 2D optical sections of T cell nuclei showed no significant enlargements, 3D volumetric measurements were necessary to show the significant differences between NLPHL and THRLBCL. We thereby anticipate that ADOPT quantification of multiparametric, immunofluorescence volumetric data of cells could not only potentially assess the functional state of T cells through nuclear measurements, but also help identify cell diseased states.…”