Experience in manufacturing a haploidentical biomedical cell product enriched with regulatory T-lymphocytes

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Abstract

Graft-versus-host disease (GVHD) remains the main life-threatening immunologic complication of hematopoietic stem cell transplantation. Despite modern pharmacological approaches for preventing and treating GVHD, there remains a need for new approaches to cure GVHD. Currently, more and more clinical experience is emerging globally in the field of using regulatory T-cell (Treg) therapies for the treatment of refractory GVHD. Manufacturing cell products for Treg therapies has a wide range of protocol variations. We have developed an approach of Treg manufacturing for cell therapy and present data from our experience in manufacturing a haploidentical Treg cell product by combining CD25+ immunomagnetic selection with closed system flowbased cell sorting methods. The study was approved by the Independent Ethics Committee and the Scientific Council of the Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology. The study describes the processes of producing 9 Treg cell products. According to the manufacturing protocol described here, it is possible to obtain a cell product that meets the quality control requirements necessary for approval for clinical use. Quality control includes the assessment of the cell composition, viability, and microbiological safety of the product, and is performed at all major stages of production. The final cell product is characterized by consistently high levels of FoxP3-expressing Treg (median: 98%), with a median cell viability of 99.1%, and has a high potential for functional efficacy. Thus, the protocol for producing Treg cell products by combining CD25+ immunomagnetic selection with flow cytometry-based cell sorting methods can be used for the clinical treatment of GVHD.

About the authors

V. A. Vedmedskaia

The Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology of Ministry of Healthcare of the Russian Federation

Email: viktoria.zubachenko@dgoi.ru
ORCID iD: 0000-0001-7247-4844
SPIN-code: 7313-8498

Victoria A. Vedmedskaya, a junior researcher at the Laboratory of Transplantation Immunology and Immunotherapy of Hemoblastoses

1 Samory Mashela St., Moscow 117997, Russia

Russian Federation

D. E. Pershin

The Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology of Ministry of Healthcare of the Russian Federation

Email: dimprsh@icloud.com
ORCID iD: 0000-0002-6148-7209

Moscow

Russian Federation

M. S. Fadeeva

The Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology of Ministry of Healthcare of the Russian Federation

Email: mailgram@inbox.ru
ORCID iD: 0000-0002-6553-2505

Moscow

Russian Federation

T. A. Sozonova

The Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology of Ministry of Healthcare of the Russian Federation

Email: tania_heurly@mail.ru
ORCID iD: 0009-0004-0057-2727

Moscow

Russian Federation

E. A. Malakhova

The Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology of Ministry of Healthcare of the Russian Federation

Email: mallahovka@yandex.ru
ORCID iD: 0000-0002-7334-0706

Moscow

Russian Federation

E. A. Kulakovskaya

The Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology of Ministry of Healthcare of the Russian Federation

Email: alenakulakovskaya@gmail.com
ORCID iD: 0000-0002-9639-2779

Moscow

Russian Federation

O. B. Lodoeva

The Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology of Ministry of Healthcare of the Russian Federation

Email: lodoeva.oyuna@gmail.com
ORCID iD: 0000-0002-2874-0014

Moscow

Russian Federation

E. Ya. Musaeva

The Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology of Ministry of Healthcare of the Russian Federation

Email: e.m.69777@gmail.com
ORCID iD: 0000-0003-0581-9472

Moscow

Russian Federation

Ya. O. Muzalevskiy

The Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology of Ministry of Healthcare of the Russian Federation

Email: fdg99@mail.ru
ORCID iD: 0000-0003-3513-8299

Moscow

Russian Federation

A. S. Kazachenok

The Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology of Ministry of Healthcare of the Russian Federation

Email: alexeykazachenok@gmail.com
ORCID iD: 0000-0003-0497-9175

Moscow

Russian Federation

D. S. Osipova

The Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology of Ministry of Healthcare of the Russian Federation

Email: d_ossipova@mail.ru
ORCID iD: 0000-0002-9968-9332

Moscow

Russian Federation

E. A. Badrin

The Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology of Ministry of Healthcare of the Russian Federation

Email: evgeny.badrin@fccho-moscow.ru
ORCID iD: 0000-0002-8678-9705

Moscow

Russian Federation

V. E. Belchikov

The Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology of Ministry of Healthcare of the Russian Federation

Email: vladbelchikov@mail.ru
ORCID iD: 0009-0001-7491-6906

Moscow

Russian Federation

A. K. Melkova

The Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology of Ministry of Healthcare of the Russian Federation

Email: a.melkova@fccho-moscow.ru
ORCID iD: 0009-0000-3801-0931

Moscow

Russian Federation

L. N. Shelihova

The Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology of Ministry of Healthcare of the Russian Federation

Email: lshelihova@gmail.com
ORCID iD: 0000-0003-0520-5630

Moscow

Russian Federation

D. N. Balashov

The Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology of Ministry of Healthcare of the Russian Federation

Author for correspondence.
Email: bala8@yandex.ru
ORCID iD: 0000-0003-2689-0569

Moscow

Russian Federation

M. A. Maschan

The Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology of Ministry of Healthcare of the Russian Federation

Email: mmaschan@yandex.ru
ORCID iD: 0000-0003-1735-0093

Moscow

Russian Federation

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Copyright (c) 2025 Vedmedskaia V.A., Pershin D.E., Fadeeva M.S., Sozonova T.A., Malakhova E.A., Kulakovskaya E.A., Lodoeva O.B., Musaeva E.Y., Muzalevskiy Y.O., Kazachenok A.S., Osipova D.S., Badrin E.A., Belchikov V.E., Melkova A.K., Shelihova L.N., Balashov D.N., Maschan M.A.

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