PMC9552184_01
Histopathology specimens from patient 1. (A) Liver biopsy showing active diffuse lymphocytic inflammation. (B) Epstein-Barr encoding region (EBER) in situ hybridization in inflammatory lymphocytic infiltrate. (C) Colon biopsy showing intense lymphocytic and polymorphic inflammatory infiltrate at the lamina propria and neutrophilic crypt micro-abscesses. (D) EBER positive in inflammatory lymphocytic infiltrate in the lamina propria and crypta. (E) Ileum biopsy showing moderate lymphocytic and polymorphic inflammatory infiltrate at the lamina propria and neutrophilic crypt microabscesses. (F) EBER positive in inflammatory lymphocytic infiltrate in the lamina propria.
fimmu-13-959733-g001.jpg
PMC9552184_01_fimmu-13-959733-g001.jpg
f1
CD3+CD8+ T cells ( Figure 1 ). Colonoscopy revealed pancolitis with ulcers ( Figure 2 ), and histopathology showed chronic active enterocolitis associated with A timeline of this patient's evolution and treatment can be found in  Supplementary Materials  as  Figure S1 .
Endoscopic features. (A) Ascending colon with edema, erythema, and multiple fibrin-covered ulcers, different sizes between 1 and 5 mm, friability; (B) Transverse colon with multiple fibrin-covered ulcers of 3 mm, patchy obliteration of vascular pattern; (C) Descending colon presence of deeper ulcers, with mild raise edge; (D) Rectum superficial small ulcers <5mm.
fimmu-13-959733-g002.jpg
PMC9552184_01_fimmu-13-959733-g002.jpg
f2
(A) CD3+ T-cells lymphoproliferation, resting and after phytohemagglutinin (PHA) stimulus. This test was performed before the HSCT. The patient shows an impaired lymphoproliferation of PHA-stimulated CD3+ T cells. (B) CD4+ and CD8+ cells subsets, naive (CD45RA+ and CCR7+), central memory (TCM, CD45RA- and CCR7+), effector memory (TEM, CD45RA-and CCR7-), and CD45RA+ effector memory cells (TEMRA, CD45RA+ and CCR7-). The patient has decreased TCM TEM CD8+ T-cells and expansion of TEMRA CD4+ T-cells. (C) CD57+ expression in CD4+ and CD8+ T-cells. Patient has increased senescent CD4+ CD57+ T-cells.
fimmu-13-959733-g003.jpg
PMC9552184_01_fimmu-13-959733-g003.jpg
f3
Further immunological workup revealed an impaired lymphoproliferation of phytohemagglutinin (PHA)-stimulated CD3+ T cells, ( Figure 3A ). Moreover, the patient lymphocyte subsets were shown to have decreased central (CD45RA- CCR7+) and effector (CD45RA- CCR7-) memory CD8+ T-cells, with expanded senescent CD4+ CD57+ T-cells ( Figures 3B, C ).
Flow cytometry from patient 1 shows (A) Diminished degranulation in Natural Killer (NK) cells from the patient (P) as compared to healthy control (HC). (B) Abnormal cell surface marker expressions in NK subsets cells from the patient as compared to healthy control, suggesting impaired NK cell differentiation.
fimmu-13-959733-g004.jpg
PMC9552184_01_fimmu-13-959733-g004.jpg
f4
NK cell function was normal based on degranulation assays ( Figure 4A ). In contrast, the expression of various cell surface markers was abnormal in NK cells from the patient as compared to healthy control, suggesting impaired NK cell differentiation ( Figure 4B ).
Lymphocyte subsets for B and Treg cells from patient 1, after failed HSCT with mixed chimerism. Plasmablasts and Memory B cells are low, as are Tregs.
fimmu-13-959733-g005.jpg
PMC9552184_01_fimmu-13-959733-g005.jpg
f5
Flow cytometry for B cell subsets and Tregs found decreased plasmablasts, memory B cells, and low Tregs ( Figure 5 ).
Family trees (A) and Sanger electropherograms (B) show de novo heterozygous missense variants in exons 4 and 7 of NFAT5, affecting Transcription activation and DNA binding domains of NFAT5 (C), in two patients with EBV-susceptibility.
fimmu-13-959733-g006.jpg
PMC9552184_02_fimmu-13-959733-g006.jpg
f6
Unfortunately, after the patient's death, the family was not willing to undergo family segregation studies. ( Figure 6 ).
PMC9552184_02
Family trees (A) and Sanger electropherograms (B) show de novo heterozygous missense variants in exons 4 and 7 of NFAT5, affecting Transcription activation and DNA binding domains of NFAT5 (C), in two patients with EBV-susceptibility.
fimmu-13-959733-g006.jpg
PMC9552184_02_fimmu-13-959733-g006.jpg
f6
Unfortunately, after the patient's death, the family was not willing to undergo family segregation studies. ( Figure 6 ).