Diagnostic assay to assist clinical decisions for unclassified severe combined immune deficiency - PubMed (original) (raw)

Diagnostic assay to assist clinical decisions for unclassified severe combined immune deficiency

Panojot Bifsha et al. Blood Adv. 2020.

Abstract

  1. 3D organoid T-cell differentiation from a few hundred peripheral blood CD34+ cells was successfully achieved.
  2. 3D organoid T-cell differentiation could help physicians distinguish intrinsic from extrinsic defects underlying a clinical SCID phenotype.

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Conflict of interest statement

Conflict-of-interest disclosure: Cultured thymus tissue implantation (RVT-802) is an investigational product implanted into patients under an Investigational New Drug application with the US Food and Drug Administration. M.L.M. is the “sponsor” of the investigations and developed the technology for cultured thymus tissue implantation. Duke has licensed the technology to Enzyvant Therapeutics. M.L.M. and Duke have received royalties from Enzyvant. Portions of M.L.M.’s and her research team’s salaries are being paid by funding from Enzyvant. If the technology is commercially successful in the future, M.L.M. and Duke may benefit financially. The remaining authors declare no competing financial interests.

Figures

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Graphical abstract

Figure 1.

Figure 1.

Abnormal in vitro T-cell differentiation of PB-CD34 + cells with intrinsic deficiency. (A) A total of 1500 CD34+ cells from PB of an IL2RG/γc SCID patient and a control (cord blood) were differentiated in 3D culture for 3 weeks. (B) A total of 500 CD34+ cells from PB of a RAG (complete deficiency) SCID patient and control (cord blood) were differentiated in 3D culture for 5 weeks. (C) A total of 600 CD34+ cells from PB of a RAG (partial deficiency; Omenn syndrome) SCID patient and control (PB of healthy adult donor) were differentiated in 3D culture for 5 weeks. Displayed events are gated from single cells, negative for 7-AAD and positive for human CD45. Percentages among CD45+7-AAD−CD34−CD19− cells (left column) and CD45+7-AAD− cells (2 right columns) are indicated in inlets. FSC-A, forward scatter area.

Figure 2.

Figure 2.

Normal in vitro T-cell differentiation of PB-CD34 + cells from SCID patients with thymic defects and ADA deficiency. (A) A total of 5000 CD34+ cells from PB of a TBX1 SCID patient and control (cord blood) were differentiated in 3D culture for 5 weeks. (B) A total of 1900 CD34+ cells from PB of a DiGeorge syndrome SCID patient and control (PB of a healthy adult donor) were differentiated in 3D culture for 5 weeks. (C) A total of 2000 CD34+ cells from PB of an ADA SCID patient and control (PB of a healthy adult donor) were differentiated in 3D culture for 3 weeks. Displayed events are gated from single cells, negative for 7-AAD and positive for human CD45. Percentages among CD45+7-AAD−CD34−CD19− cells (left column) and CD45+7-AAD− cells (2 right columns) are indicated in inlets.

References

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