Abstract
Recent literature suggests conventional flow cytometric (FCM) immunophenotyping complemented by Ki-67 FCM assessment as a reliable tool to classify canine lymphomas. Ki-67 expression assessed by FCM is rarely reported in canine lymphoma cases and reference data for normal canine lymph nodes are missing. Moreover, nothing is known about the Ki-67 expression within the occasionally observed remnant cell population within the gates of normal lymphocytes in lymphoma cases. Aim of this study was to compare flow cytometric Ki-67 expression of lymphocyte populations from normal canine lymph nodes, lymphoma cells from World-Health-Organisation (WHO) classified lymphoma patient samples and their neighboring normal remnant cell population. Cryopreserved lymphocyte cell suspensions from normal lymph nodes from eight dogs free of lymphoma served as reference material. Fourteen cases diagnosed by cytology, FCM, clonality testing, histopathology including immunohistochemistry consisting of 10 DLBCL, 1 MZL, 1 PTCL and 2 TZL showed a residual small lymphocyte population and were investigated. The Ki-67 expression in normal canine lymphoid tissue was 3.19 ± 2.17%. Mean Ki-67 expression in the malignant cell populations was 41 ± 24.36%. Ki-67 positivity was 12.34 ± 10.68% in the residual physiologic lymphocyte population, which otherwise exhibited a physiologic immunophenotype pattern. This ratio was equivalent (n = 3) or lower (n = 11) than the Ki-67 expression of the malignant cell population within the sample. This is the first report of FCM derived Ki-67 expression combined with immunophenotype patterns in normal canine lymph nodes, compared with lymphoma cell populations and residual normal cell populations of lymphoma cases diagnosed by state of the art technology.
Highlights
Ki-67 is a nuclear protein strictly associated with cellular proliferation due to its expression during all active phases of the cell cycle (G1, S, G2, and M-phase), and absence in quiescent (G0) cells [1, 2]
For diagnosis of Diffuse large B-cell lymphoma (DLBCL), cytology combined with flow cytometry (FCM) and simultaneous Ki-67 assessment from fine needle aspirates are propagated as diagnostic gold standard allowing prediction of treatment response replacing conventional grading [1]
The cases were classified by cytology, FCM, clonality testing, histopathology including immunohistochemistry and consisted of 10 DLBCL, 1 marginal zone lymphoma (MZL), 1 Peripheral T-cell lymphoma (PTCL) and 2 T-Zone lymphoma (TZL)
Summary
Ki-67 is a nuclear protein strictly associated with cellular proliferation due to its expression during all active phases of the cell cycle (G1, S, G2, and M-phase), and absence in quiescent (G0) cells [1, 2]. A time consuming and error prone task as mitotic activity is unevenly distributed within the tumor so that observation and sampling errors are method inherent [21]. For diagnosis of Diffuse large B-cell lymphoma (DLBCL), cytology combined with flow cytometry (FCM) and simultaneous Ki-67 assessment from fine needle aspirates are propagated as diagnostic gold standard allowing prediction of treatment response replacing conventional grading [1]. This spares the patients a more invasive sampling such as lymph node excision or wedge biopsies and avoids the time consuming, costly and inherently error prone histopathologic procedures
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