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4CLINICAL AND LABORATORY- DIAGNOSTIC INDICATORS BY DINT OF FLOW CYTOFLUOROMETRY AND THE FREQUENCY PREVALENCE OF ACUTE LEUKEMIA AMONG
PATIENTS CHILDREN
Baratova D.,
ANMSO "Eurasian Center of Oncohematology, Immunology and Therapy"
Saint-Petersburg, Russia ANO "National register of hematopoietic stem cells Kirghizia"
Saint-Petersburg, Russia Mamatysaeva U.
ANO "National register of hematopoietic stem cells Kirghizia" Saint-Petersburg, Russia
SI "National Center of Oncology and Hematology" at the Ministry of Health Kyrgyz Republic(Kirghizia), Bishkek
DOI: 10.5281/zenodo.7672995
Abstract
The article presents the results of immunotyping (IFT) studies by dint of the flowing cytofluorimetry method and determined frequency the of elicitation of acute leukemia among sick children in the Kyrgyz Republic( Kirghizia).
In the diagnosis of acute leukemia with by dint of flow cytofluorometry, it is necessary to evaluate the im-munophenotypical characteristics of tumor (blast) cells and determine their directions of myeloid, B and T linear or rare variants of vaguely linearity.
According to the results of our studies, in typing immunophenotyping by dint of flow cytofluorimetry in prevalence disease revealed acute lymphoblastic leukemia in 68% of cases, acute myeloid leukemia in 29% of cases and acute leukemia of unclear linearity in 3% of cases.
It is noted that, to the frequency of prevalence of acute leukemia among sick children of residents of the Kyrgyz Republic((Kirghizia), elicitation the incidence of acute myeloid leukemia among child females in 38% of cases, among child males in 23% of cases, and acute lymphoblastic leukemia occurs among child females in 62% of cases and in child males in 77% of cases.
However, with timely diagnosis of acute leukemia with the determination of the linearity variant and the selection of PCTs, and with the presence of an HLA-identical healthy donor, haploidentical, closely related hem-atopoietic stem cell transplantation (brother, sister, parent or placental blood), in particular in children, gives the best results.
Keywords: flow cytofluorometry, acute leukemia, children's, kirghiz nationality.
INTRODUCTION:
Today, acute leukemia occurs in all developing and developed countries, and remains one of the global health problems. With the correct installation diagnosis, it is important to achieve a good result from targeted therapy.
Acute leukemia - is acute malignant disease of the blood system, with defeats at the level of deterministic genus-parent stem cells or early cellular predecessor, characterized by the availability of blast cells in the bone marrow puncture or in peripheral blood from 20% and more [1].
As the authors note[1], when diagnosing acute leukemia and at other oncohematological diseases, cytogenetic, morphological, molecular genetic studies remain a powerful tool for apply in a wide clinical practice, but also for a detailed study of bone marrow cells and determining the linear the direction variant, flow cytofluometry , undoubtedly, has its great advantages.
Immunophenotyping using flow cytofluometry makes it possible to evaluate the immunophenotypic characteristics of tumor cells, the direction of myeloid, B and T-linear or rare variants of unclear linearity and after each stage of therapy to determine the minimumresidual disease (MRD), when diagnosing acute leukemia and other oncohematological malignant dis-eases[1;6;7].
However, for the diagnostic purpose of determining the linear affiliation of tumor cells, sets of antibodies recommended by International Consensus are used Bethesda[10], the Euroflow Consortium[9], the International Expert Group of the European Leukemia Network (ELN)[5], as well as for the differential diagnosis of the classification of the European Group for the Immunological Characterization of Leukemia(EGIL)[3] and the World Health Organization(WHO) [8].
In the disease of acute leukemia with the determination of the linearity variant and the timely selection of targeted therapy with the achievement of a therapeutic effect, it is necessary to determine the minimum residual disease, and perform allogeneic hematopoietic stem cell transplantation according to the indications, in particular in children give the best results. Conducting a high-technological method of haploidentical ther-apy(brother, sister, parent or placental blood), closely related or unrelated hematopoietic stem cell transplantation, allows in the presence of an HLA-identical healthy donor[2].
The purpose of our study is a timely diagnostic analysis of the variant of the immunophenotype of tumor (blast) cell linearity and to determine the prevalence of acute leukemia in patients children in the Kyr-gyz Republic(Kirghizia).
MATERIALS AND METHODS:
The study group from November 2016 to December 2022 years, included 103 children patients (child patients female -42, child patients male -61), children of kirghiz nationality- 77 patienst children and residents of the Russian-speaking population of the Kyrgyz Republic (Kirghizia) - 26 patients children, with acute lymphoblastic leukemia-70 patients of children (child patients female - 25, child patients male -45)- of which 52 children patients of kirghiz nationality and residents of the Russian-speaking population of the Kyrgyz Re-public(Kirghizia) - 18 children's patients(child patients female -3, child patients male -15).With acute myeloid leukemia-30are children's patients(child patients female -16, child patients male -14) of them are children patients of kirghiz nationality -24 (child patients fe-male-14 and child patients male-10) and residents of the Russian-speaking population (mixed nation and different nationalities) Kyrgyz Republic (Kirghizia) -6 patients of children, including children (child patients female -2, child patients male-4), with acute leukemia of unclear linearity of the Russian-speaking population of the Kyrgyz Republic (Kirghizia)-3patients of chil-dren(child patients female-1, child patients male-2), all patients children aged from 1,5 years to16 years, and citizens of the Kyrgyz Republic (Kirghizia), who were passing examined at the Department of Pediatric Oncology of the National Center Oncology and Hematol-ogy Department of Health of the Kyrgyz Republic (Kirghizia) and in the Department of Pediatric Hematology Osh interregional clinical children's hospital in Osh, in St. Petersburg(Russia) consult doctor-hematologists Eurasian Center of Oncohematology, Immunology and Therapy.
Method immunophenotyping by dint of flowing cytofluorometry conducted the first time and research immunophenotyping was conducted in city Bishkek, Kyrgyz Republic (Kirghizia)[1].
METHOD BY DINT OF FLOWING CYTOFLUOROMETRY:
The material for the study is the bone marrow. Patients made puncture of the sternum. For obtain a qualitative result, the obtained analysis should not be with impurity blood and not destroy the cells during the testsample preparation to immunophenotyping. Immuno-phenotyping of leukemia (blast) cells performed on a flow cytofluometer Cytomics FC500 (Beckman Coulter, USA) using monoclonal antibodies Beckman Coulter.
Statistical processing of the results included the analysis of standard criteria X2-square, which was used to assess the significance of differences in the occurrence of certain characteristics between the control group and the test group. Determination of the "p", the corresponding value found. X2-square performed considering one degree of freedom.
All mathematical calculations and statistical analysis of the overall study was performed using a personal computer using the package application programs for spreadsheets -"Microsoft - Excel M version 7.0, for Windows 95, for Windows-based 2010, Statistica-5.
RESULTS AND DISCUSSION:
At the conducted our studies patient children, with diagnosis of acute leukemia was established on the basis of a detailed collection of anamnesis, clinical data and a comprehensive row of general and special laboratory-diagnostic indicators.
It also is taken into account and the importance for us the doctors-clinician in making a diagnosis, in children-patients with acute leukemia, mother's place of work (connection with chemical and toxic substances, dyes) the area of residence with a high radiation background and with bad ecology, taking prohibited medi-cation(drugs), does it have hereditary genetic diseases, age and concomitant diseases in parents were carried out, independent delivery or cesarean section, the weight and height of the child at birth. And currently in what living conditions the child live after birth.
Prenatal and postnatal periods are of great importance: obstetric anamnesis of mother and child, the frequency of ultrasound examinations of the fetus, given the high radiation background -long-distance flights by plane in the first 3rd trimester, infectious and acute respiratory viral diseases during pregnancy, allergic diseases, frequent stressful situations, blood transfusion.
In the early stages of the disease, may occur in the form of bacterial, viral (especially herpes), protozoal, fungal infection, mainly nasopharyngeal lesions, with catarrhal phenomena of redness, sore throat, on the skin of the body in the form of petechiae, perianal area and when contacting a doctor, the doctor first draws attention to the symptoms of hematopoietic depression, bleeding gums, lesion of a group of lymphatic nodes, enlarged liver, spleen, central nervous system, skin-signs of infiltration of organs by leukemic cells, high indicators of elevated body temperature, expression of anemic, intoxication syndromes. In the study of a general blood test, where the presence of blast cells of various fluctuations on the hemogram, where it is possible to suspect about disease of acute leukemia.
The diagnosis of acute leukemia was established according to the recommendations of the World Health Organization(WHO, 2017) [8] and the classification of the French-American-British Cooperative Group (FAB,1985) [4].
We clinicians in accepted for the final diagnostic limit of acute leukemia disease on the basis of key standard criteria, including the classical triad: clinical manifestations, tumor (blast) cells in peripheral blood from several to large quantities, in the bone marrow from 20% or more.
And also in each child patient, the clinical condition, concomitant diseases and laboratory diagnostic, morphological, immunogenetic cytogenetic indicators were taken into account, followed by an assessment of their prognostic significance: age, gender, a variant of the course of the linearity of the disease.
To determine the prevalence of acute leukemia among sick children, according to the results of (clinical data, laboratory-diagnostic indicators) and also studies by dint of flow cytofluorometry,weredistributed on the cell linearity variants(Figure1).
The prevalence of acute leukemia in the Kyrgyz Republic 3% (Kirghizia)
29%
68%
acute lymphoblastic leukemia in patients children
I acute myeloid leukemia in patients children
acute leukemia of unclear linearity in patients children
Figure1.Comparative characteristics of the variants course of acute leukemia and the frequency ofprevalence in
the Kyrgyz Republic among patients children.
At the comparative characteristic on the frequency of prevalence, as can be seen from the presented Figure 1, among patients children of residents of the Kyrgyz Republic(Kirghizia), acute lymphoblastic leukemia occurs in
of cases and acute leukemia of unclear linearity in 3% of cases.
In the significance of the differences, have reliable statistics differences, where p>0,0005.
68% of cases, acute myeloid leukemia in 29%
100% 90% 80% 70% 60% 50°% 40% 30% 20% 10% 0%
in patients children female
in patients children male
acute myeloid leukemia
acute lymphoblastic leukemia
Figure 2. The frequency ofprevalence of acute leukemia among female and male children-patients in the Kyrgyz
Republic (Kirghizia).
According to the results of our data from the presented Figure 2, it can be seen that in frequency prevalence among patients childrens the detection of acute myeloid leukemia in child females in 38% of cases, among child males in 23% of cases, and acute lympho-blastic leukemia occurs among child females in 62 % of cases and child males in 77% of cases.
The significance of the differences have statistically reliable differences, where p>0.005.
CONCLUSIONS:
Thus, it is necessary to conduct a timely clinical diagnostic study, determine the linearity variant by dint of flowing cytofluorometry, identify the features of the course disease and, after the final diagnosis, prescribe targeted treatment. At the diagnosing the disease of acute leukemia, despite studying the detailed characteristics of tumor (blast) cells using the flow cytofluorim-etry method, it allows to identify a characteristic im-munophenotype, but in order to confirm the diagnosis, it is necessary to conduct cytogenetic, molecular genetic and morphological studies in parallel.
According to the frequency of prevalence, acute leukemia occurs among both female and male children. Acute lymphoblastic leukemia is most often detected among child male patients, and acute myeloid leukemia among child female patients. Acute lymphoblastic leukemia prevails among patients children in the Kyrgyz Republic(Kirghizia).
After installation diagnosis and determination of the direction of the linearity variant, timely selection of polychemotherapy is necessary, when remission is achieved, hematopoietic stem cell transplantation, in the presence of an HLA- identical healthy donor (brother, sister, parent or placental blood).
FINDINGS:
1. To investigate the bone marrow method by dint of flowing cytofluorometry in patient children's at the suspected acute leukemia.
2. Immunophenotyping by dint of flowing cyto-fluorimetry makes it possible to identify the characteristic linearity of the immunophenotype of a tumor (blast) cell and timely select an effective program chemotherapy.
3. In parallel conducts cytogenetic, cytochemical, morphological, molecular- genetic studies.
References
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