Evidence map›Paper›PMID 42458494›Full record

ArticleJournal of translational medicine2026

Development of a 3-day manufacturing method to generate CD19-CD20-CD22 trispecific CAR T-cells from whole blood.

Isabella Vignola, Michaela Prochazkova, Lipei Shao, Tatyana Fuksenko, Sarmila Sarkar, Hong Lei, Jianjian Jin, Ying Xiong, Yanping Xie, Ibeawuchi Oparaocha and 10 more

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Journal of translational medicine, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

1 citing paper in PubMed.

  1. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

20 authors.

Isabella VignolaCenter for Cellular Engineering, Department of Transfusion Medicine, National Institutes of Health, Bethesda, MD, USA.
Michaela ProchazkovaCenter for Cellular Engineering, Department of Transfusion Medicine, National Institutes of Health, Bethesda, MD, USA.
Lipei ShaoCenter for Cellular Engineering, Department of Transfusion Medicine, National Institutes of Health, Bethesda, MD, USA.
Tatyana FuksenkoCenter for Cellular Engineering, Department of Transfusion Medicine, National Institutes of Health, Bethesda, MD, USA.
Sarmila SarkarCenter for Cellular Engineering, Department of Transfusion Medicine, National Institutes of Health, Bethesda, MD, USA.
Hong LeiCenter for Cellular Engineering, Department of Transfusion Medicine, National Institutes of Health, Bethesda, MD, USA.
Jianjian JinCenter for Cellular Engineering, Department of Transfusion Medicine, National Institutes of Health, Bethesda, MD, USA.
Ying XiongCaring Cross, Gaithersburg, MD, USA.
Yanping XieCaring Cross, Gaithersburg, MD, USA.
Ibeawuchi OparaochaCaring Cross, Gaithersburg, MD, USA.
Oxana SlessarevaCaring Cross, Gaithersburg, MD, USA.
Megan ForrestCaring Cross, Gaithersburg, MD, USA.
Rimas OrentasCaring Cross, Gaithersburg, MD, USA.
Zhongyu ZhuCaring Cross, Gaithersburg, MD, USA.
Boro DropulicCaring Cross, Gaithersburg, MD, USA.
Ping JinCenter for Cellular Engineering, Department of Transfusion Medicine, National Institutes of Health, Bethesda, MD, USA.
Robert P SomervilleCenter for Cellular Engineering, Department of Transfusion Medicine, National Institutes of Health, Bethesda, MD, USA.
David F StroncekCenter for Cellular Engineering, Department of Transfusion Medicine, National Institutes of Health, Bethesda, MD, USA.
Hannah W SongCenter for Cellular Engineering, Department of Transfusion Medicine, National Institutes of Health, Bethesda, MD, USA.
Steven L HighfillCenter for Cellular Engineering, Department of Transfusion Medicine, National Institutes of Health, Bethesda, MD, USA. Steven.Highfill@NIH.Gov.ORCID 0000-0003-0135-0961

Funding

Intramural NIH HHS Z99 CL999999NIH HHS Z99CL999999
6 · The paper itself

Abstract

backgroundChimeric antigen receptor (CAR) T-cell therapy has transformed the treatment landscape for many hematological malignancies. However, high relapse rates and limited accessibility remain significant challenges. We developed a 3-day streamlined process to address these limitations.

methodsT-cells were isolated from whole blood collected from healthy donors via an automated density gradient separation that included a T-isopure™ antibody cocktail, which isolates T-cells through negative selection. T-cells were activated then transduced with a lentiviral vector encoding a trispecific CAR. T-cells were cultured in G-Rex vessels and harvested at day 3 or day 7 for analysis. CAR expression and T-cell phenotype were assessed by flow cytometry and gene expression analysis. Functional activity was evaluated by measuring cytotoxicity and cytokine secretion following co-culture with target cell lines.

resultsThe T-isopure isolation enriched CD3

conclusionThis study establishes a rapid, GMP-compliant method for manufacturing polyfunctional, CAR T-cells directly from whole blood. The workflow outlined here achieved a potent, phenotypically favorable CAR T-cell product without compromising viability or cytotoxic function. Compared to the standard 7-day method, the 3-day approach resulted in expression of genes associated with a more stem-like phenotype while reducing manufacturing time and cost. This method may provide a practical alternative for decentralized CAR T-cell manufacturing, particularly in resource-limited settings.

Indexed as

Antigens, CD19Antigens, CD20Receptors, Chimeric AntigenT-LymphocytesCytokinesHumansPhenotypeTime FactorsAntigens, CD19Antigens, CD20CytokinesReceptors, Chimeric AntigenCell manufacturingCellular therapyChimeric antigen receptorNegative selectionWhole blood

Identifiers

PMID42458494
PMCPMC13378159

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LicenceCC BY
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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.