Immunohistochemical analysis indicates that the anatomical location of B-cell non-Hodgkin's lymphoma is determined by differentially expressed chemokine receptors, sphingosine-1-phosphate receptors and integrins.
BACKGROUND
The aim of this study was to elucidate the mechanisms responsible for the location of B-cell non-Hodgkin's lymphoma (B-NHL) at different anatomical sites. We speculated that the malignant B cells in these disorders have the potential for trafficking between blood and secondary lymphoid organs (SLO) or extranodal sites and that their preferential accumulation at different locations is governed by the expression of key molecules that regulate the trafficking of normal lymphocytes.


METHODS
Biopsy or blood samples from 91 cases of B-NHL affecting SLO (n = 27), ocular adnexae (n = 51) or blood (n = 13) were analysed by immunohistochemistry or flow cytometry for the expression of the following molecules: CCR7, CCL21 and αL (required for the entry of normal lymphocytes into SLO); CXCR4, CXCL12 and α4 (required for entry into extranodal sites); CXCR5, CXCL13 and S1PR2 (required for tissue retention); S1PR1 and S1PR3 (required for egress into the blood). The expression of each of these molecules was then related to anatomical location and histological subtype.


RESULTS
The expression of motility/adhesion molecules varied widely between individual patient samples and correlated much more strongly with anatomical location than with histological subtype. SLO lymphomas [comprising 10 follicular lymphoma (FL), 8 diffuse large B-cell lymphoma (DLBCL), 4 mantle-cell lymphoma (MCL) and 5 marginal-zone lymphoma (MZL)] were characterised by pronounced over-expression of S1PR2, suggesting that the malignant cells in these lymphomas are actively retained at the site of clonal expansion. In contrast, the malignant B cells in ocular adnexal lymphomas (10 FL, 9 DLBCL, 4 MCL and 28 MZL) expressed a profile of molecules suggesting a dynamic process of trafficking involving not only tissue retention but also egress via S1PR3 and homing back to extranodal sites via CXCR4/CXCL12 and α4. Finally, leukaemic lymphomas (6 FL, 5 MCL and 2 MZL) were characterised by aberrant expression of the egress receptor S1PR1 and low expression of molecules required for tissue entry/retention.


CONCLUSIONS
In summary, our study strongly suggests that anatomical location in B-NHL is governed by the differential expression of specific adhesion/motility molecules. This novel observation has important implications for therapeutic strategies that aim to disrupt protective micro-environmental interactions.
Department of Molecular and Clinical Cancer Medicine, University of Liverpool, Liverpool, England.
Stephen
S
Middle
Department of Molecular and Clinical Cancer Medicine, University of Liverpool, Liverpool, England.
Sarah E
SE
Coupland
Medical Physics and Clinical Engineering, Royal Liverpool University Hospital, Liverpool, England.
Azzam
A
Taktak
ORLAU, RJAH Orthopaedic hospital NHS Foundation Trust, Oswestry, England.
Victoria
V
Kidgell
Department of Molecular and Clinical Cancer Medicine, University of Liverpool, Liverpool, England.
Joseph R
JR
Slupsky
Department of Molecular and Clinical Cancer Medicine, University of Liverpool, Liverpool, England.
Andrew R
AR
Pettitt
Department of Molecular and Clinical Cancer Medicine, University of Liverpool, Liverpool, England.
Kathleen J
KJ
Till
B-NHL; Chemokine; Egress; Homing; Integrin; Lymphoma; Microenvironment; S1P receptors