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. 2021 Apr 22;12:650427. doi: 10.3389/fimmu.2021.650427

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Copyright © 2021 Tang, Shimada and Ikegaki

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Clinical relevance of CD4 CTL in high-risk neuroblastoma. (A) The study cohort and its UH subgroups. The study cohort is composed of 176 high-risk neuroblastoma specimens (40, 41), which include the Null group and EUH groups (MYC-driven, TERT over-expression, and ATRX loss). The proportion of each subset was estimated based on expression levels of MYCN, MYC, TERT and ATRX. (B) HLA-E is highly expressed in high-risk neuroblastoma. HLA-E expression was correlated with PHOX2B expression (a marker of neuroblastoma) in the study cohort. All tumors were PHOX2B positive and co-expressed HLA-E. (C) The effect of HLA-E expression on survival of high-risk neuroblastoma. High HLA-E expression was associated with better outcome of high-risk neuroblastoma. Survival of high-risk neuroblastoma patients with high or low expression of HLA-E was analyzed by the R2: Genomics Analysis and Visualization Platform (http://r2.amc.nl). (D–F) The effect of NKG2E, CD4 and CD8 expression on survival of high-risk neuroblastoma. (D) High NKG2E and (E) high CD4 expressions were associated with prolonged survival of high-risk neuroblastoma. Difference in High expression (Blue) vs. Low expression (Red) was statistically significant up to 130 months after diagnosis as indicated by the arrows in (D, E). (F) No association between CD8 expression and disease outcome was found. CD8 expression is represented by CD8 β chain (CD8B). (G) Correlation between CD4 and CD3E expressions in the high-risk neuroblastoma. The expression of CD4 and CD3E were highly correlated each other, suggesting the presence of tumor-infiltrating CD4+ T cells. CD3E (encoding the CD3ε chain) expression represents CD3 expression. The horizontal bar represents the cutoff value to separate the cohort into high and low CD4 subsets, which were used in the survival analysis shown in (E). (H) The effect of CRTAM expression on high-risk neuroblastoma. High CRTAM expression, encoding a CTL activating receptor, was associated with prolonged survival of high-risk neuroblastoma. (I) High-risk neuroblastoma expresses both HLA-E and CADM1. All the high-risk tumors examined expressed both HLA-E and CADM1. CADM1, encoding the CRTAM ligand, expressed at high levels in high-risk neuroblastoma. CADM1 expression also showed a trend of being associated with better survival (p=0.096) (not shown). Unit of expression levels is expressed as Reads Per Million (RPM). Expression levels of genes shown in the figures were expressed as log2 of RPM.