anti samsn1 antibody  (Boster Bio)

Journal: medRxiv

Article Title: SAMSN1 causes sepsis immunosuppression by inducing macrophages to express coinhibitory molecules that causes T cell exhaustion via KEAP1-NRF2 signaling

doi: 10.1101/2024.10.03.24314746

Figure Lengend Snippet: (a) The flow chart shows that proteins bound to SAMSN1 are first precipitated by IP, and then the transcription factors that bind to SAMSN1 and change under LPS stimulation are analyzed by mass spectrometry. Among the transcription factors that are upregulated after LPS stimulation, JUNB, ZFP36, NFKIBZ and KEAP1 can be found in the public datasets for RNA-Seq data after knocking them out. (b) GSE216352 dataset shows the expression of co-inhibitory molecules CD48, CD86, and CEACAM1 in WT and KEAP1-KO RAW264.7 cells. (c) Real-time RT-PCR detection of CD48/CD86/CEACAM1 mRNA levels after WT or SAMSN1-KO cells were stimulated with LPS. (d) Real-time RT-PCR detection of CD48/CD86/CEACAM1 mRNA levels in WT cells stimulated with LPS after treatment with the KEAP1 inhibitor ML385 (5μM for 1h). (e) Real-time RT-PCR detection of CD48/CD86/CEACAM1 mRNA levels in SAMSN1-KO cells stimulated with LPS after treatment with the agonist KI696 (20μM KI696 for 24h). (f) IP-WB assay showed that the specific binding of SAMSN1 and KEAP1 increased when WT cells were stimulated with LPS. (g) IP-WB assay detected the binding of KEAP1 and NRF2 in LPS-stimulated WT or SAMSN1-KO cells. Significance in (b-e) was assessed by 2-tailed Welch’s t test .

Article Snippet: Primary antibodies: Anti-p-ZAP70 antibody (CST Cat. # 2717); anti-ZAP70 antibody (CST Cat. # 2705); anti-p-LCK antibody (CST Cat. #2751); anti-LCK antibody (CST Cat. # 2984); anti-Actin antibody (HUABIO); anti-Tubulin antibody (HUABIO); KEAP1; anti-SAMSN1 antibody (AtaGenix).

Techniques: Mass Spectrometry, RNA Sequencing Assay, Expressing, Quantitative RT-PCR, Binding Assay

Journal: medRxiv

Article Title: SAMSN1 causes sepsis immunosuppression by inducing macrophages to express coinhibitory molecules that causes T cell exhaustion via KEAP1-NRF2 signaling

doi: 10.1101/2024.10.03.24314746

Figure Lengend Snippet: Schematic diagram: In the resting state, KEAP1 and NRF2 form a transcription complex in the cytoplasm of monocyte-macrophages and are in an inhibitory state. When sepsis occurs, the number of monocyte-macrophages increases significantly, in which SAMSN1 is highly expressed. SAMSN1 binds to KEAP1, causing NRF2 to dissociate from the complex and translocate into the nucleus, promoting the transcription of co-inhibitory molecules CD48/CD86/CEACAM1. Through cell-cell contact, these co-inhibitory molecules bind to corresponding receptors 2B4/TIM4/CD152 on the surface of T cells, causing T cell inhibition and exhaustion. High expression of SAMSN1 also switches macrophages into a suppressive phenotype, manifested by decreased phagocytic ability and ability to clear bacteria in the blood. Blockade of SAMSN1 by gene knockout or monoclonal antibody keeps KEAP1-NRF2 in a complex state, preventing NRF2 from entering the nucleus and promoting the transcription of downstream co-inhibitory molecules, thereby failing to inhibit T cell activation. Blockade of SAMSN1 also switches macrophages to a pro-immune phenotype with enhanced phagocytic ability, resulting in reduced organ injuries and increased survival rate.

Article Snippet: Primary antibodies: Anti-p-ZAP70 antibody (CST Cat. # 2717); anti-ZAP70 antibody (CST Cat. # 2705); anti-p-LCK antibody (CST Cat. #2751); anti-LCK antibody (CST Cat. # 2984); anti-Actin antibody (HUABIO); anti-Tubulin antibody (HUABIO); KEAP1; anti-SAMSN1 antibody (AtaGenix).

Techniques: Inhibition, Expressing, Bacteria, Gene Knockout, Activation Assay