case_images: PMC10728670_01
case_images: Timeline with relevant data from the patient's history. WT, Wilms' tumor; CT, chemotherapy; RT, radiotherapy; US, ultrasound; IVIG,intravenous immunoglobuline; MRI, magnetic resonance imaging.
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case_images: Complete blood count performed as part of the oncological follow-up revealed progressive lymphopenia from the age of 10 years ( Figure 1 ).
case_images: (A) Family pedigree with the inheritance pattern. TRIM37 duplication and deletion are showed. (B) Sanger sequencing of TRIM37 gene revealed the presence of the monoallelic NM_001320987.3 c.2357dupA (p. Asp786GlufsTer7), a duplication leading to a stop codon seven amino acids downstream. (C) Schematic representation of the TRIM37 gene and TRIM37 protein. The positions of mutations and deletions (colored horizontal lines) reported in literature in association with an immunological phenotype are shown in green (Jobic et al) and blue (Bruzzaniti et al) colors. In red is depicted the molecular lesions found in the patient reported in this paper. (D) Flow cytometry plots of peripheral blood mononucleate cells from the patient and two healthy controls. CD4+ and CD8+ cells subsets gated on CD3+ cells and T-cell subsets. (E) Comparison of T-cell differentiation subsets in patient and controls. HC1, healthy control 1; HCs, healthy control 1 and healthy control 2; PT, Mulibrey patient; TEMRA, terminally differentiated effector memory T cells; TEM, T-effector memory; TCM, T-central memory. Black arrow indicates the proband.
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case_images: Figures 2D, E ). Flow cytometry-based analysis of T-cell receptor (TCR) Vbeta repertoires demonstrated a normal distribution among CD4+ Vbeta families.
case_images: PMC10728670_02
case_images: (A) Family pedigree with the inheritance pattern. TRIM37 duplication and deletion are showed. (B) Sanger sequencing of TRIM37 gene revealed the presence of the monoallelic NM_001320987.3 c.2357dupA (p. Asp786GlufsTer7), a duplication leading to a stop codon seven amino acids downstream. (C) Schematic representation of the TRIM37 gene and TRIM37 protein. The positions of mutations and deletions (colored horizontal lines) reported in literature in association with an immunological phenotype are shown in green (Jobic et al) and blue (Bruzzaniti et al) colors. In red is depicted the molecular lesions found in the patient reported in this paper. (D) Flow cytometry plots of peripheral blood mononucleate cells from the patient and two healthy controls. CD4+ and CD8+ cells subsets gated on CD3+ cells and T-cell subsets. (E) Comparison of T-cell differentiation subsets in patient and controls. HC1, healthy control 1; HCs, healthy control 1 and healthy control 2; PT, Mulibrey patient; TEMRA, terminally differentiated effector memory T cells; TEM, T-effector memory; TCM, T-central memory. Black arrow indicates the proband.
case_images: fimmu-14-1303251-g002.jpg
case_images: PMC10728670_02_fimmu-14-1303251-g002.jpg
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case_images: Figures 2D, E ). Flow cytometry-based analysis of T-cell receptor (TCR) Vbeta repertoires demonstrated a normal distribution among CD4+ Vbeta families.
case_images: (A) CD4+ proliferation assay after 4 days stimulation with phytohemagglutinin 1 mcg/ml or antiCD3 and anti-CD28 1 mcg/ml with or without IL2 10 ng/ml. (B) Exemplary cell trace violet dilution after 4 days stimulation with PHA 1 mcg/ml. (C) CD25-expression level in CD4+ and CD8+ cells after 4 days stimulation with anti-CD3 1 mcg/ml and anti-CD28 1 mcg/ml. (D) Six-week thymocyte development assessed by artificial thymic organoid generated by culturing DLL4-expressing stromal cell line (MS5-hDLL4) with CD34+ cells isolated from peripheral blood cells of the patient compared to healthy control. MFI, mean fluorescence intensity; HC1, healthy control 1; HCs, healthy control 1 and 2; PT, Mulibrey patient; PHA, phytohemagglutinin.
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case_images: f3
case_images: Proliferation was clearly impaired after PHA and anti-CD3/anti-CD28 stimulation, and addition of interleukin-2 (IL-2) did not rescue proliferation ( Figures 3A, B ). Surface marker analysis upon in-vitro activation showed increased expression of the activation marker CD25 when cells were stimulated with anti-CD3/anti-CD28 after 48h, as compared to healthy controls ( Figure 3C ). In-vitro T-cell development at 6 weeks of culture appeared normal and comparable to healthy controls ( Figure 3D ), excluding a hematopoietic-intrinsic defect in T-cell development.