Fluorescence in situ hybridization (FISH) analysis of primary ocular adnexal MALT lymphoma

BACKGROUND: It remains unknown whether primary ocular adnexal extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma) is a homogeneous entity, as there are few reports of the results of cytogenetic or molecular analyses of these tumors. METHODS: We performed interphase fluorescence in situ hybridization (FISH) analysis to detect translocations and aneuploidy in 34 cases of primary ocular adnexal MALT lymphoma, and reviewed the histopathological findings. Correlations between the results of FISH analysis, the histopathological features and the clinical data were also analyzed. RESULTS: Among the 34 cases, FISH analysis revealed t(14;18)(q32;q21) in one case, trisomy 3 in 21 cases (62%), and trisomy 18 in 16 cases (47%). The cases with trisomy 18 had significantly more prominent lymphoepithelial lesions (LELs) and less nodularity in the tumors. In regard to the clinical correlations, tumors with trisomy 18 were observed predominantly in females and younger patients; also, in the majority of the cases, the tumor was of conjunctival origin. All the cases with recurrence showed trisomy 18 in the tumor. CONCLUSION: Primary ocular adnexal MALT lymphoma is a significantly heterogeneous entity. Cases with trisomy 18 may have unique clinicopathological features

Immunostaining using an anti-F4/80 antibody.

<p>(A) Representative images of fracture hematoma at day 3. Scale bar = 200 μm. <i>DAP12</i>^{<i>–/–</i>} mice showed less fluorescence from F4/80-positive cells in fracture hematoma. (B) Quantitative analysis of F4/80-positive cells. The number of F4/80-positive cells was significantly decreased in <i>DAP12</i>^{<i>–/–</i>} mice than in B6 mice. *<i>p</i><0.05.</p

mRNA expression of osteogenic, chondrogenic and proinflammatory genes.

<p>(A) Expression levels of <i>Col1a1</i>, <i>Col2a1</i>, and <i>Col10a1</i> in the fracture callus. <i>DAP12</i>^{<i>–/–</i>} mice demonstrated a delayed increase in <i>Col1a1</i> gene expression. <i>DAP12</i>^{<i>–/–</i>} mice showed significantly lower expression of <i>Col1a1</i> at day 7 and significantly higher expression at day 14. In <i>DAP12</i>^{<i>–/–</i>} mice, the expression of <i>Col2a1</i> and <i>Col10a1</i> was significantly lower at day 5 and significantly higher at day 14. <i>DAP12</i>^{<i>–/–</i>} mice showed delayed decreases in <i>Col2a1</i> and <i>Col10a1</i> gene expression. (B) Expression levels of <i>Tnf</i>, <i>Il1b</i>, and <i>Il6</i> in fracture hematomas. The expression of <i>Tnf</i> was significantly lower in <i>DAP12</i>^{<i>–/–</i>} mice at days 3 and 5. There were no significant differences in the expression of <i>Il1b</i> or <i>Il6</i>. *<i>p</i><0.05, **<i>p</i><0.01, ***<i>p</i><0.001.</p

Micro-CT image analysis of fracture callus at day 14 in B6 and <i>DAP12</i>^{<i>–/–</i>} mice.

<p>(A) Representative axial images of fracture callus in B6 and <i>DAP12</i>^{<i>–/–</i>} mice. Dense trabecular structure and deteriorated cortical shell formation on the surface in <i>DAP12</i>^{<i>–/–</i>} mice compared with B6 mice. (B) Structural parameter analysis of fracture callus of B6 and <i>DAP12</i>^{<i>–/–</i>} mice. Trabecular thickness was significantly larger in <i>DAP12</i>^{<i>–/–</i>} mice than in B6 mice, and the percentage of new cortical shell area was significantly lower in <i>DAP12</i>^{<i>–/–</i>} mice. There were no significant differences in TV, BV/TV, Tb.N or Tb.Sp. *<i>p</i><0.05.</p

Histological images of fracture healing.

<p>(A) Alcian blue-stained sections. Scale bar = 500 μm. <i>DAP12</i>^{<i>–/–</i>} mice showed more residual cartilaginous tissue and delayed woven bone formation (arrow) at day 10. Substantially impaired cortical shell formation on the callus surface was observed in <i>DAP12</i>^{<i>–/–</i>} mice at day 14 (arrowhead). <i>DAP12</i>^{<i>–/–</i>} mice showed residual cartilage matrix in the peripheral callus, less original cortex absorption and delayed bone remodeling at day 28. (B) Quantitative histomorphometric analysis of the fractured callus in B6 and <i>DAP12</i>^{<i>–/–</i>} mice. There was no significant difference in callus volume between <i>DAP12</i>^{<i>–/–</i>} and B6 mice. Hematoma area was significantly larger in <i>DAP12</i>^{<i>–/–</i>} mice than in B6 mice at day 7. The newly formed bone area was significantly smaller in <i>DAP12</i>^{<i>–/–</i>} mice than in B6 mice at days 7 and 10. Cartilage area was significantly larger in <i>DAP12</i>^{<i>–/–</i>} mice than in B6 mice at day 10. *<i>p</i><0.05, **<i>p</i><0.01.</p

Radiographic images of fractured tibiae in B6 and <i>DAP12</i>^{<i>–/–</i>} mice.

<p><i>DAP12</i>^{<i>–/–</i>} mice showed lower callus volume and radiolucency at days 21 and 28 compared with B6 mice (arrow head).</p