Comment on "ALK is a therapeutic target for lethal sepsis"

Abstract

Physiologically relevant ALK (anaplastic lymphoma kinase) expression was not detected in human and mouse monocytes and macrophages, suggesting that the effects of bioactive compounds on stimulator of interferon genes (STING) activation may not depend on ALK.

Fig. 1.. Undetectable ALK expression in monocytes and macrophages.

(A) ALK mRNA expression in tissues according to the Genotype-Tissue Expression (GTEx) project. (B) ALK expression in murine cell lines [top: SP8 anti-ALK antibody (1:1000); bottom: F12 anti-ALK antibody ( 1:100)]. Positive controls: 293T cells transfected with mouse Alk (293T_ALK) and EML4-ALK-expressing TeCla1 cells. Negative controls: untransfected 293T and KP1233 murine KRAS^G12D-driven NSCLC cells. Arrows indicate the full-length (f.l.) ALK or the EML4-ALK fusion. (C) ALK expression in human cell lines [top: D5F3 anti-ALK antibody (1:1000); bottom: F-12 anti-ALK antibody (1:100)]. Positive controls: human neuroblastoma cell lines SK-N-BE and SH-SY5Y. Negative controls: human B-lymphoma Raji and Ramos cell lines. Arrows indicate the full-length (f.l.) ALK or the cleaved ALK. Asterisk indicates the non-specific band detected by the F-12 antibody. (D) ALK expression in human cell lines [left: D5F3 anti-ALK (1:1000); right: F-12 anti-ALK antibody (1:100)]. Positive control: THP-1 cells transduced with human ALK receptor (THP-1_ALK). Negative control: THP-1 cells transduced with empty retroviral vector (THP-1_vector). H3122 and H2228 NSCLC cell lines carry the EML4-ALK (ex13-ex20) and the EML4-ALK (ex6-ex20) rearrangements that generate fusion proteins of 130 kDa and 90 kDa, respectively. Karpas-299 is an NPM-ALK-rearranged cell line that generates a fusion protein of 80 kDa. Positive controls: SK-N-BE and SH-SY5Y. Arrows indicate the full-length (f.l.) ALK or the cleaved ALK. Asterisk indicates the non-specific band detected by the F-12 antibody. Top panels: short exposure; Bottom panels: long exposure (E) Immunohistochemistry performed on an ALK-rearranged NSCLC case (D5F3 anti-ALK antibody) and on an ALCL case (4C5B8 anti-ALK antibody). Tumor cells shown by black arrows; macrophages shown by red arrows. Scale bar indicates 50 µm.

Fig. 2.. Undetectable ALK mRNA expression in Raji and THP-1 cell lines.

ALK mRNA expression in different tumor cell lines according to two different datasets in the Cancer Cell Line Encyclopedia (CCLE) (https://portals.broadinstitute.org/ccle). (A) RNA sequencing data; (B) Gene expression profiling data. Raji, THP-1 and the ALK-positive neuroblastoma cell lines SH-SY5Y and SK-N-BE are indicated by arrows.