Introduction of the method of flow cytometry in the diagnosis of chronic lymphoproliferative diseases of the medical center west kazakhstan marat ospanov medical university

Oncohematological pathology has been occupying an important place in the structure of cancer incidence in the Aktobe region for many years. The hematology department, the Department of transplantation and cell therapy functions on the basis of the Medical Center West Kazakhstan Marat Ospanov Medical University. Over the past years, the total number of newly identified and observed hematological oncological patients has been gradually increasing, for 2020, the number of cases of blood disease amounted to 640 cases, of which 92 cases are chronic lymphoproliferative diseases (CLPD) [1].

Over the past 10-15 years, significant progress has been made in the diagnosis of oncohematological diseases, which made it possible to identify new diagnostic markers. Nevertheless, despite the increasing penetration of molecular methods into oncohematology, morphological analysis has its niche and remains an important primary link in the diagnosis of leukemia in the MC of theWKMU . Laboratory medicine is the leading one and forms the basis for evidencebased medicine, so in 2020 it was decided to introduce the flow cytometry method for the diagnosis of leukemia in most cases of CLP, without which it is impossible today to exist oncohematology, transplantology, and clinical decision-making on a larger number of diseases.

Diagnostic tests are carried out on the Beckman Coulter Navios analyzer. The analyzer is capable of measuring small-angle (up to 3 angles) and lateral light scattering, as well as registering fluorescence at ten wavelengths, using 3 different lasers, thus providing the possibility of simultaneous investigation of many parameters.

Today there is a working panel designed in compatibility with the similarity of monoclanal markers and large ones with certain diagnoses. In acute leukemia, a panel for the determination of monoclonal antibodies CD15, CD64, CD44, CD11c, CD34, CD117, CD13, CD 33, CD14, CD56, CD45, chronic lymphoproliferative diseases CD103, CD19, CD200, CD5, SD7, CD3, CD4, CD8, CD79a, CD25, CD38, CD138, CD20.

Flow cytofluorimetry is successfully used in the diagnosis of malignant lymphoproliferative diseases. However, sometimes there are atypical situations that are difficult to interpret, which serves as the basis for the search for new markers with differential diagnostic significance. Therefore, we included in the panel an analysis of the expression of CD200 in patients with malignant B-cell lymphoproliferative diseases [2] for differential diagnosis.

The developed diagnostic algorithms make it possible to successfully differentiate the majority of immunomorphological variants of B-cell CLPD. At the first stage, groups of tumors with CD19+CD5+, CD19+10+ and CD19+CD5–CD10-phenotypes are traditionally distinguished. The next step in patients with CD19+CD5+ expression is to analyze the expression of CD23, CD22, CD79b, CD20 antigens on the surface membrane, which allows diagnosing typical CLL and lymphoma from mantle zone cells (LMZ) [3-6]. However, there are observations with atypical expression when diagnosis is difficult. For example, cases of atypically low expression of CD5, CD23, CD43 and pronounced expression of CD20, CD22, CD79b have been described in CLL. In addition, there are observations with CD23 expression, decrease and absence of CD5 expression in LMZ. It should be noted that, quite often, other lymphomas (not CLL and not LMZ) fall into the group with CD19+CD5+ expression, in particular lymphomas from marginal zone cells (LMZ), lymphoplasmocytic lymphoma, hairy cell leukemia (HCL), in which CD5 expression is uncharacteristic [6]. In such situations, the diagnosis can be verified by a combination of histological, immunohistochemical and cytogenetic studies. Thus, although the diagnosis of B-cell CLP by flow cytometry based on the study of blood and bone marrow lymphoid cells has advantages, in some cases it is not possible to avoid more invasive, time-consuming and timeconsuming methods. It follows from this that the diagnostic panel needs improvement. One of the markers recommended for inclusion in the diagnostic panel by many authors is CD200.

At the moment, research work is underway on the topic: "Analysis of the main and small populations of peripheral blood lymphocytes of patients with lymphoproliferative diseases using the method of multicolored flow cytofluorometry", where the hypothesis of the study is: monoclonal antibodies CD200, CD103, CD 11c in the main multicolored panel are highly informative markers for differential diagnosis of various forms of lymphoproliferative diseases.

Research objectives:

1. To explore the major and minor populations of lymphocytes of patients with lymphoproliferative diseases and to establish the phenotypic features of expression of cell surface markers CD 200, CD103, CD11c on b lymphocytes of chronic lymphocytic leukemia (CLL), hairy cell leukemia (HCL), lymphoma cells of the mantle zone (LMZ), lymphoma marginal zone of the spleen (LMZS).

2. To compare the results of immunophenotyping with morphological research methods for the final diagnosis. list of sources:

  2. Yu.V. Mirolyubova., E.A. Stadnik., T.S. Nikulina., V.V.Strugov., K Andreeva., Yu.V. Virts., R.V. Grozov., Ayu Zaritskaya. Clinical oncohematology. 2017;10(2):169–75
  3. Lugovskaya S.A.Kisilichina D.G., Pochtar M.E. et al. New markers (CD160, CD200, LAIR-1) in the diagnosis of B-cell lymphoproliferative diseases. Clinical oncohematology. 2013;6(1):45-52.
  4. Kupryshina N.A., Tupitsyn N.N. Flow cytometry in hematology. Part II. Fundamentals and innovations in the diagnosis of chronic lymphocytic leukemia. Clinical oncohematology. 2012;5(4):349–54.
  5. Stadnik E.A., Strugov V.V., Virts Yu.V., Zaritsky A.Yu. Chronic lymphocytic leukemia. Recommendations for diagnosis and treatment. Translational medicine. 2012;17:104-15.
  6. Sverdlov Sh., Campo E., Harris N.L. et al., ed. WHO Classifies tumors of hematopoietic and lymphoid tissues. 4th edition. Lyon: IARC Press; 2008. 4. Konke T., Wittmann V.K., Soter D. et al. Proposal For A New Assessment System For The Diagnosis Of CLL. Blood. 2013; 122(21):47-5599 (Abstracts of the plenary sessions).
Year: 2021
City: Shymkent
Category: Medicine