Keyword: CD3

ABSTRACT: Introduction: CAR-T cell therapy is the newest and most effective treatment method for resistant forms of B-cell hematological malignancies; however, it has until now remained unavailable in Kazakhstan. The first and critically important step in the manufacturing of CAR-T products is apheresis-based leukapheresis — a procedure that determines the quality of the starting cellular material for genetic modification of T lymphocytes. In the Republic of Kazakhstan, this procedure was performed within the framework of a national scientific and technical program for the implementation of CAR-T therapy.
Objective: To study the parameters of apheresis-based leukapheresis and to characterize the cellular composition of the obtained mononuclear cell concentrate in order to assess its suitability as starting material for the manufacturing of CAR-T products.
Materials and Methods:  At the Department of Cell Technologies of the Scientific and Production Center of Transfusiology (Astana), 12 leukapheresis procedures were performed in 7 healthy donors and 5 patients with oncohematological diseases. An automated separator Spectra Optia (Terumo BCT, USA) was used with the Mononuclear Cell Collection program. The volume of processed blood, procedure duration, product cellularity, CD3⁺ T-cell content, and sterility were assessed.
Results: All 12 procedures were completed without complications. The mean volume of processed blood was 9549.5 mL (1.0–2.6 blood volumes), and the mean duration was 230.8 min. Cell yield varied: total leukocyte count ranged from 6.5×10⁹ to 32.1×10⁹ (mean 14.4±8.0×10⁹), and CD3⁺ T-lymphocyte content ranged from 129.1×10⁷ to 1629.7×10⁷ cells per dose. Products from all participants were sterile and deemed suitable for CAR-T cell manufacturing. Differences were noted between the donor and patient groups; however, due to the small sample size, statistical analysis was not performed.
Conclusions: Apheresis-based leukapheresis yields a cellular concentrate that fully meets the requirements for initiating the manufacturing of CAR-T products. This work represents the first standardized protocol in Kazakhstan for obtaining clinically significant cellular material for CAR-T cell manufacturing and lays the foundation for the development of national quality criteria.
Funding: Program-Targeted Funding of the Ministry of Health of the Republic of Kazakhstan BR25293293 "Implementation of chimeric antigen receptor (CAR)-T cell therapy technology for hematological tumors into practical healthcare."

ABSTRACT: Introduction: Under conditions of limited availability of immunohistochemical antibodies, the key challenge is the rational use of a minimal panel that allows confirmation of the lymphoid nature of the tumor, determination of B- or T-cell lineage, and selection of the further direction of investigation.
Objective: To evaluate the feasibility of a stepwise diagnostic algorithm for B-cell lymphomas using a limited antibody panel.
Materials and Methods; A retrospective analysis of 139 cases of lymphoproliferative diseases was conducted. At the first stage, CD45, CD20, CD3, PanCK, and Ki-67 were used. CD45 was used to confirm the lymphoid nature of the process, PanCK to exclude epithelial tumors, CD20 and CD3 to determine B- or T-cell lineage, and Ki-67 to assess proliferative activity. Additional antibodies were ordered based on morphological and immunophenotypic indications, taking into account their actual availability.
Results: A B-cell phenotype was established in 100 of 139 cases (71.9%), Hodgkin lymphoma in 35 (25.2%), and T-cell lymphomas in 4 (2.9%). Among the 100 B-cell lymphomas, in 32 cases (32.0%), the diagnosis was formulated at the level of B-cell lymphoma without precise nosological subclassification. Diffuse large B-cell lymphoma/large B-cell lymphoma was diagnosed in 36 cases (36.0%), follicular lymphoma in 10 (10.0%), small/middle B-cell lymphomas, including SLL/CLL, in 9 (9.0%), marginal zone lymphomas/MALT-type in 7 (7.0%), Burkitt lymphoma/highly aggressive B-cell lymphoma in 4 (4.0%), and mantle cell lymphoma in 2 (2.0%). The high proportion of diagnoses without complete subclassification reflects the limited availability of additional markers, whereas the basic panel allowed lineage determination and identification of the need for a second diagnostic stage.
Conclusion: The CD45/CD20/CD3/PanCK/Ki-67 panel is a practical first step in the diagnosis of lymphoproliferative diseases under resource-constrained conditions. Expansion of the panel should be performed in a targeted manner, based on morphology and the results of the first stage. In the absence of the necessary antibodies, establishing the B- or T-cell phenotype is a justifiable level of diagnostic conclusion and allows avoidance of unjustified nosological verification.

ABSTRACT: Introduction: POEMS syndrome is a rare paraneoplastic disorder associated with plasma cell dyscrasia and characterized by a combination of organomegaly, peripheral polyneuropathy, endocrinopathies, osteosclerotic lesions, monoclonal shedding, and typical skin changes. Diagnosis is challenging due to the nonspecific clinical presentation and minimal bone marrow infiltration. Currently, treatment approaches remain nonstandard, and the use of anti-CD38 therapy is limited.
Objective: To demonstrate the complexity and duration of diagnosis of POEMS syndrome and evaluate the clinical effect of anti-CD38 therapy.
Materials And Methods: A retrospective analysis of clinical, laboratory, and imaging data from a 38-year-old patient seen at the National Cancer Research Center was conducted. Diagnostic evaluation included serum enzyme-linked immunosorbent assay, vascular endothelial growth factor (VEGF) levels, PET/CT, bone marrow examination, and assessment of compliance with the Dispenzieri 2019 criteria. Treatment efficacy was assessed based on clinical symptoms and laboratory findings.
Results: The patient was diagnosed with polyneuropathy, organomegaly, extravascular fluid overload, endocrinopathy, monoclonal IgG-λ secretion, and significantly elevated VEGF levels. Minimal plasma cell infiltration was noted in the bone marrow. POEMS syndrome was diagnosed. Treatment with Daratumumab-Lenalidomide-Dexamethasone (Dara-RD) resulted in significant clinical improvement, decreased edema, and stabilization of laboratory parameters.
Conclusion: This case demonstrates the complexity of diagnosing POEMS syndrome with minimal bone marrow infiltration, severe systemic manifestations, and problems with long-term diagnostic verification, lasting for 5 years (from 2019 to 2025). Daratumumab-Lenalidomide-Dexamethasone (DaraRD) therapy has demonstrated significant clinical benefit and can be considered a progressive treatment option for severe forms of the disease.