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Isolating And Immunostaining Lymphocytes and Dendritic Cells from Murine Peyer's Patches
Published on: March 17, 2013
Dendritic reticulum cell-related immunostaining for laminin in follicular and diffuse B-cell lymphomas
1Division of Pathology, Centro di Riferimento Oncologico, Aviano, Italy.
Insights
This study reveals that laminin and follicular dendritic reticulum cells (DRCs) are closely associated in B-cell lymphomas. Their presence correlates with nodularity, suggesting a trapping function in lymphoma microenvironments.
Area of Science:
- Immunohistochemistry
- Oncology
- Cell Biology
Background:
- Follicular dendritic reticulum cells (DRCs) and laminin play roles in lymph node microenvironments.
- Non-Hodgkin's lymphomas (NHLs) exhibit altered microenvironmental structures.
Purpose of the Study:
- To comparatively analyze the distribution of laminin and DRCs in B-cell NHLs.
- To investigate the relationship between laminin, DRCs, and lymphoma nodularity.
Main Methods:
- Immunohistocytochemistry and immunofluorescence techniques were used.
- Analysis included 29 B-cell NHL cases and 5 reactive lymph nodes.
- Single and double labeling assays with anti-laminin and DRC-1 antibodies were performed.
Main Results:
- Laminin showed a punctate-granular pattern in reactive germinal centers and neoplastic nodules, paralleling DRC distribution.
- In diffuse lymphoma areas, laminin was present only where DRC nests were observed.
- Laminin and DRC-1 positivity suggested co-localization on the same cells.
Conclusions:
- A rich DRC meshwork and associated laminin correlate with nodularity in reactive and neoplastic conditions.
- Reduced DRCs and laminin are linked to a loss of nodularity in lymphoma.
- DRCs may exert a trapping function in the presence of laminin within the lymphoma microenvironment.
Abstract:
We performed a comparative immunohistocytochemical study of the distribution patterns of laminin and follicular dendritic reticulum cells (DRCs) within their follicular microenvironment in both nodular or diffuse B-cell non Hodgkin's lymphomas (NHLs). Twenty nine cases of immunophenotypically diagnosed B-cell NHLs (19 of follicular center cell origin-FCCL- and 10 of the diffuse well differentiated lymphocytic type-WDLL-) and five reactive lymph nodes with follicular hyperplasia were analyzed by immunoperoxidase and immunofluorescence techniques. Serial frozen sections and cytospin preparations were tested either with single antibodies anti laminin and DRC-1, or paired reagents in double labeling immunofluorescence. Our results indicated consistently that within both the reactive germinal centers and the neoplastic nodules of FCCL laminin immunostaining visualized a punctate-granular pattern apart from the linear vascular basement membrane positivity. Double immunofluorescence assay demonstrated that there was a close parallelism between this laminin staining pattern and DRC-1 distribution showing a well developed DRCs meshwork; in the diffuse tumour areas of both FCCL and WDLL, laminin immunoreactivity was found only in those cases in which nests of DRCs were observed. Double immunofluorescence studies performed on cytospin preparations demonstrated that the groups of cells containing DRC-1 positive cells, contained a positivity for laminin, although within the cell the staining for DRC-1 was intense and diffuse, while that for laminin was granular and more sparse. Our results suggested that these laminin and DRC-1 positive reactive sites may be present on the same cells. Since the reduction in number or loss of both DRCs and their related immunostaining for laminin within the microenvironment was consistently associated with a loss of nodularity by lymphoma cells, whereas nodularity in reactive and neoplastic conditions was associated with a rich DRCs meshwork and the related laminin immunostaining, a trapping function of DRCs exercised in the presence of laminin should be considered.
