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. 2024 Jan;36(1):86-94.
doi: 10.1177/10406387231204873. Epub 2023 Oct 13.

Characterization of CD3+/CD20+ canine large-cell lymphoma

Cory M Howard  1   2 , Steffanie Anderson  2 , Bonnie Harrington  3
Affiliations

Characterization of CD3+/CD20+ canine large-cell lymphoma

Cory M Howard et al. J Vet Diagn Invest. 2024 Jan.

Abstract

Immunophenotyping of canine large-cell lymphoma (LCL) for B-cell and T-cell surface antigens is commonly performed to better predict the clinical outcome. Expression of surface antigen CD3 is associated with T-cell malignancies; surface antigen CD20 is expressed on B cells. However, a small subset of canine LCLs expresses both CD3 and CD20 (CD3+/CD20+); this form of lymphoma remains poorly defined at the molecular level. In a retrospective study, we aimed to better characterize immunophenotypic properties and antigen receptor clonality of CD3+/CD20+ LCL. We selected formalin-fixed, paraffin-embedded tissues from 10 cases of CD3+/CD20+ LCL and breed-matched controls of peripheral large T-cell lymphoma (PTCL) and diffuse large B-cell lymphoma (DLBCL). Using PCR for antigen receptor rearrangement (PARR), we identified monoclonal T-cell receptor gamma (TCRγ) rearrangements in all CD3+/CD20+ cases. Three of 10 cases had monoclonal rearrangements in the immunoglobulin heavy chain (IgH), supportive of cross-lineage rearrangement. There was no significant difference in the frequency of antigen receptor rearrangement between CD3+/CD20+ and PTCL cases. In comparison with DLBCL, CD3+/CD20+ LCL had TCRγ rearrangement more frequently and IgH rearrangement less frequently, respectively. Immunolabeling of the B-cell marker PAX5 occurred less frequently in all CD3+/CD20+ LCL cases compared to the DLBCL controls. Immunolabeling for BCL-2 was robust, regardless of immunophenotype. Nuclear Ki67 positivity was variable in CD3+/CD20+ cases, indicating a heterogeneity in proliferation. Overall, cases of canine CD3+/CD20+ LCL had properties similar to PTCL, suggesting a similar histogenesis of these 2 subsets.

Keywords: BCL-2; CD20; CD3; DLBCL; PARR; PAX5; PTCL; canine; large-cell lymphoma.

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Conflict of interest statement

Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Figures

Figure 1.
Figure 1.
Representative H&E stains and CD3 and CD20 immunolabeling images from CD3+/CD20+ large-cell lymphoma (LCL) and control cases. Case 10, CD3+/CD20+ LCL: A = H&E; B = CD3; C = CD20. Case 20, peripheral large T-cell lymphoma: D = H&E; E = CD3; F = CD20. Case 28, diffuse large B-cell lymphoma: G = H&E; H = CD3; I = CD20. Bars = 50 µm.
Figure 2.
Figure 2.
Antigen receptor clonality in CD3+/CD20+ canine large-cell lymphoma. A. Representative images from the amplification of immunoglobulin heavy chain (IgH). C1–8 and E10–12 correspond to the lane identification in the QIAxcel advanced capillary electrophoresis cartridge. B. Representative images from the amplification of T-cell receptor gamma (TCRγ). A1–A12 correspond to the lane identification in the QIAxcel advanced capillary electrophoresis cartridge.
Figure 3.
Figure 3.
Expression of PAX5 and BCL-2 in CD3+/CD20+ canine large-cell lymphoma (LCL). Representative images of immunolabeling of PAX5 and BCL-2. Case 10, CD3+/CD20+ LCL: A = PAX5; B = BCL-2. Case 28, peripheral large T-cell lymphoma: C = PAX5; D = BCL-2. Case 11, diffuse large B-cell lymphoma: E = PAX5; F = BCL-2. Bars = 50 µm.
Figure 4.
Figure 4.
Ki67 immunolabeling index in CD3+/CD20+ canine large-cell lymphoma (LCL). Representative images of immunolabeling of Ki67 from CD3+/CD20+ lymphoma: A = case 1; CD3+/CD20+ LCL; B = case 4, CD3+/CD20+ LCL; C = case 11, peripheral large T-cell lymphoma; D = case 28, diffuse large B-cell lymphoma. Bars = 50 µm. E. Quantitation of % Ki67 positivity. Results were compared using a one-way ANOVA and a Tukey HSD (*p ≤ 0.05). The box-and-whisker plot indicates the minimum, first quartile, median, x– (“x”), third quartile, and maximum of each dataset.
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