case_images: PMC8448282_01
case_images: Histologic evaluations: (A-E) Hematoxylin-eosin of benign lymph node biopsy of P1 and P2. Figures at low magnification shows: (A,C,D) large and irregularly expanded "naked" GC with ill-defined outlines, loss of the mantel zone and polarization, monocytoid B-cell hyperplasia, numerous large histocyte and tingible bodies (A,C:4x, D: 5x); (A) the arrow shows area of sclerosis around the follicle. Figures at higher magnification shows: (B) absence of mantel zone surrounding the germinal center (40x); (E) foci of necrosis (20x). (F-N) Hematoxylin-eosin of malignant biopsy of P1 and P2. (F,G,I) infiltration by polymorphous DLBCL of large bowel (F:4x, I:20x, G: 40x) and (J,K) of lympho-node (J: 4x, K:40x). (L,M) low (10x) and high (40x) magnification of lympho-node of P2 infiltered by NLPHL. Arrows show "popcorn" cells with multilobate nucleus and scant cytoplasm surrounded by rosetting activate small lymphocytes. (H, N)
  In-situ hybridization for EBER, arrow shows some positive cells (x10, in ellipse high magnification x40).
case_images: fped-09-703853-g0002.jpg
case_images: PMC8448282_01_fped-09-703853-g0002.jpg
case_images: F2
case_images: The main histologic finding was an atypical follicular hyperplasia characterized by voluminous "naked" GC with ill-defined contours, marked reduction of mantle zone and prominent monocytoid B cells reaction (Figures 2A,B). Lymph node and endoscopic duodenal mucosa biopsies revealed a diffuse large B cell lymphoma (DLBCL) (Figures 2F,G). Sporadic cells resulted EBER-positive (Figure 2H). Foci of necrosis were also present (Figures 2C-E). One year later, at 18 years old, after the recurrence of cervical lymphadenopathy, she received a diagnosis of a nodular lymphocyte-predominant Hodgkin Lymphoma (NLPHL) (Figures 2L,M). A minority of cells resulted EBER-positive (Figure 2N). The histologic examination of bowel biopsy and lymph nodes revealed a DLBCL developed in the context of lymphatic hyperplasia (Figures 2I-K).
case_images: Functional analysis. (A) Western blot analysis of p-Akt and beta-actin protein expression in T blast from P1 (APDS1), P2 (APDS2) and HD with or w/o OKT3-CD28 stimulation (10 min); (B) Percentage of apoptotic (7AAD-AnnexinV+) or necrotic (7AAD+AnnexinV +-) CD4 and CD8 gated cells from APDS1 (P1) and APDS2 (P2) patients with or w/o 48 h of OKT3-CD28 stimulation. Results were showed as fold increase calculated as ratio between patients and HD. (C)
  Ex vivo phospho-S6 Ser235-236 intracellular staining performed on whole blood in CD3+ gated lymphocytes from P1 (APDS1) and P3 and P4 (APDS2) pre and post therapy with PI3K inhibitor and a healthy donor (HD). (D) Bar plots represent pattern of CD107a expression measured by flow cytometry on gated NK cells of an APDS2 patient at two time points (30 and 34 years of age) and of HDs (n = 7, age ranging from 17 to 34 years) following 6 h culture of PBMCs (effectors, E) with and without (not stimulated, ns) K562 cell targets (T) at an E:T ratio of 10:1. The median +- IQR is reported for HDs.
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case_images: PMC8448282_01_fped-09-703853-g0003.jpg
case_images: F3
case_images: They showed a well-tolerance, improved immune phenotype (Table 1), functional test (Figure 3C), clinical symptoms and quality of life. IN P2 the NK cell cytotoxicity resulted similar to that of HD controls at two different time points (Figure 3D). Western blot of pAKT of P1 and P2 (Figure 3A) and flow cytometry analysis of Phospho S6 Ser235/236 of P1, P3, P4 (Figure 3C) showed the hyperactivation of the PI3K-AKT-mTOR signaling pathway. In P3 and P4 cytometry analysis showed a reduction of Phospho S6 Ser235/236 after the treatment with Seletalisib (Figure 3C). In vitro CD4+ and CD8+ T cells resulted more apoptotic and necrotic after stimulation with anti-CD3/CD28 respect to the unstimulated condition and the HD control (P1, P2, Figure 3B).