Flow Cytometry in the Diagnosis of Canine B-Cell Lymphoma

Riondato, Fulvio; Comazzi, S.

doi:10.3389/fvets.2021.600986

Cited by 16 publications

(13 citation statements)

References 56 publications

(54 reference statements)

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“…CD3, CD79acy) including Ki-67 expression ( 30 ). A population of smaller cells could be clearly distinguished from the malignant lymphoma cell population in the forward scatter/ side scatter (FSC/SSC) dot plot as published recently ( 31 ) ( Figures 1B,D ) ( Figures 1B,D,H,J,N,P ).…”

Section: Methodssupporting

confidence: 75%

Flow Cytometric Assessment of Ki-67 Expression in Lymphocytes From Physiologic Lymph Nodes, Lymphoma Cell Populations and Remnant Normal Cell Populations From Lymphomatous Lymph Nodes

et al. 2021

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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.

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Section: Methodssupporting

confidence: 75%

Flow Cytometric Assessment of Ki-67 Expression in Lymphocytes From Physiologic Lymph Nodes, Lymphoma Cell Populations and Remnant Normal Cell Populations From Lymphomatous Lymph Nodes

et al. 2021

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“…For diagnosis of lymphomas, however, cytologic evaluation (e.g., size and morphology of aberrantly expanded cells) is important even if a 9-color panel is utilized. As it has been reported [ 26 ], cell size measured via FSC properties by flow cytometry is useful to distinguish B-cell lymphoma cells that are larger than B cells in normal/reactive lymph node ( S6 Fig ). In case of diagnostic fine-needle aspirates, the advantage of using a 9-colour panel, compared to a standard used 2 to 4 color panel, is that only a single tube is sufficient for diagnosis, which is particularly important if low numbers of cells are aspirated.…”

Section: Discussionmentioning

confidence: 69%

Use of multi-color flow cytometry for canine immune cell characterization in cancer

et al. 2023

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Although immunotherapy is becoming a standard approach of human cancer treatment, only a small but critical fraction of patients responds to the therapy. It is therefore required to determine the sub-populations of patients who will respond to immunotherapies along with developing novel strategies to improve efficacy of anti-tumor immune reactions. Current development of novel immunotherapies relies heavily on mouse models of cancer. These models are important for better understanding of mechanisms behind tumor immune escape and investigation of novel strategies to overcome it. Nevertheless, the murine models do not necessarily represent the complexity of spontaneously occurring cancers in humans. Dogs spontaneously develop a wide range of cancer types with an intact immune system under similar environment and exposure to humans, which can serve as translational models in cancer immunotherapy research. To date though, there is still a relatively limited amount of information regarding immune cell profiles in canine cancers. One possible reason could be that there are hardly any established methods to isolate and simultaneously detect a range of immune cell types in neoplastic tissues. To date only a single manuscript describes characterization of immune cells in canine tumour tissues, concentrating solely on T-cells. Here we describe a protocol for multi-color flow cytometry to distinguish immune cell types in blood, lymph nodes, and neoplastic tissues from dogs with cancer. Our results demonstrate that a 9-color flow cytometry panel enables characterization of different cell subpopulations including myeloid cells. We also show that the panel allows detection of minor/aberrant subsets within a mixed population of cells in various neoplastic samples including blood, lymph node and solid tumors. To our knowledge, this is the first simultaneous immune cell detection panel applicable for solid tumors in dogs. This multi-color flow cytometry panel has the potential to inform future basic research focusing on immune cell functions in translational canine cancer models.

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“…When a lymphoma is suspected the target cells are lymphocytes. Based on this criteria, specific antibodies which detect certain CD ( CD3, CD5, CD4, CD8, CD34, CD21, CD45, MHC-II) are routinely used for the superficial labeling, while CyCD79b and CyCD3 are used for intracellular antigen detection (34,35). Often these antibodies are combined together and placed in the same tube in order to make possible a multiparametric analysis.…”

Section: Antibodiesmentioning

confidence: 99%

Applicability of Flow Cytometry in Identifying and Staging Lymphoma, Leukemia and Mast Cell Tumors in Dogs: An Overview

Sulce

Munga²,

Beqiraj³

et al. 2022

SVR

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Lymphomas, leukemias and mast cell tumors belong to the most important group among all neoplasms affecting dog species. Diagnosis, staging and determining the cell type involved in a specific tumor represent a challenge for researchers and clinicians, and plays a crucial role in treatment efficacy and prognostic purposes. Many different gold standard techniques such as cytology, histopathology, immunohistochemistry and cytochemistry are used to routinely diagnose and stage these tumors. In the recent years flow cytometry is becoming more applicable in veterinary medicine since a wide number of health conditions can be analyzed in a short period of time with a high accuracy. Multiparametric analysis performed by flow cytometry is considered as one of the main advantages of this technique since cell populations can be analyzed for different superficial markers at the same time. Immunophenotyping and staging of tumor cell populations performed by flow cytometry can help in reaching a confirmatory diagnosis and appropriate prognosis of the disease. Moreover, many flow cytometric results have been linked to a high prognostic relevance especially in neoplastic disorders. However, flow cytometry results are compatible and should be interpreted in compliance with data obtained by histopathology, immunohistochemistry and cytology. Key words: flow cytometry; antibodies; diagnosis; lymphoma; leukemia UPORABNOST PRETOČNE CITOMETRIJE PRI PREPOZNAVANJU IN DOLOČANJU STADIJA LIMFOMA, LEVKEMIJE IN TUMORJEV MASTOCITOV PRI PSIH – PREGLED Izvleček: Limfomi, levkemije in tumorji mastocitov so najpomembnejše skupine neoplazem, ki prizadenejo pse. Diagnostika, določanje stopenj tumorja in tipa celic v določenem tumorju predstavljajo izziv za raziskovalce in klinike in igrajo ključno vlogo pri učinkovitosti zdravljenja in postavljanju prognoze. Za rutinsko diagnosticiranje in določanje stopenj teh tumorjev se uporablja veliko različnih temeljnih metod, kot so citologija, histopatologija, imunohistokemija in citokemija. V zadnjih letih je pretočna citometrija vse bolj uporabljana metoda v veterinarski medicini, saj je mogoče v kratkem času in z visoko natančnostjo analizirati veliko število zdravstvenih stanj. Ena izmed najpomembnejših prednosti te tehnike je multiparametrična analiza, s katero lahko v populaciji celic istočasno analiziramo različne površinske označevalce. Določanje površinskih označevalcev in stopenj populacij tumorskih celic s pretočno citometrijo lahko pripomore k potrditvi diagnoze in postavitvi ustrezne prognoze bolezni. Številni rezultati pretočne citometrije so imeli pomemben prognostični pomen zlasti pri neoplastičnih obolenjih. Vendar je rezultate pretočne citometrije potrebno združiti in razlagati v skladu s podatki, pridobljenimi s histopatologijo, imunohistokemijo in citologijo. Ključne besede: pretočna citometrija; protitelesa; diagnoza; limfom; levkemija

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Flow Cytometry in the Diagnosis of Canine B-Cell Lymphoma

Cited by 16 publications

References 56 publications

Flow Cytometric Assessment of Ki-67 Expression in Lymphocytes From Physiologic Lymph Nodes, Lymphoma Cell Populations and Remnant Normal Cell Populations From Lymphomatous Lymph Nodes

Flow Cytometric Assessment of Ki-67 Expression in Lymphocytes From Physiologic Lymph Nodes, Lymphoma Cell Populations and Remnant Normal Cell Populations From Lymphomatous Lymph Nodes

Use of multi-color flow cytometry for canine immune cell characterization in cancer

Applicability of Flow Cytometry in Identifying and Staging Lymphoma, Leukemia and Mast Cell Tumors in Dogs: An Overview

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