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Generation of mouse macrophages expressing membrane-bound TNF variants with selectivity for TNFR1 or TNFR2.

Abstract：

:Tumor necrosis factor-alpha (TNF) is expressed on the cell surface as a transmembrane form (tmTNF), that can be released as a soluble form (solTNF) via proteolytic cleavage. These two types of TNF exert their biological functions by binding to one of two TNF receptors, TNFR1 or TNFR2. However, the biological function of tmTNF through these two receptors remains to be determined. Here, we generated macrophages that expressed tmTNF mutants with selectivity for either TNFR1 or TNRF2 as a tool to evaluate signaling through these receptors. Wild-type TNF (wtTNF), TNFR1-selective mutant TNF (mutTNF-R1) or TNFR2-selective mutant TNF (mutTNF-R2) were individually expressed on the TNFR1(-/-)R2(-/-) mouse macrophages (Mphi) as the tmTNF forms. tm-mutTNF-R1-expressing Mphi exhibited significant selectivity for binding to TNFR1, whereas tm-mutTNF-R2-expressing Mphi only showed a slight selectivity for binding to TNFR2. Signaling by tm-mutTNF-R1-expressing Mphi through the hTNFR2 was weaker than that of tm-wtTNF-expressing Mphi, suggesting that the binding selectivity correlated with functional selectivity. Interestingly, signaling by tm-mutTNF-R2-expressing Mphi through TNFR2 was much stronger than signaling by tm-wtTNF-expressing Mphi, whereas signaling by the corresponding soluble form was weaker than that mediated by wtTNF. These results indicate tmTNF variants might prove useful for the functional analysis of signaling through TNF receptors.

journal_name

Cytokine

journal_title

Cytokine

authors

Shibata H,Abe Y,Yoshioka Y,Nomura T,Sato M,Kayamuro H,Kawara T,Arita S,Furuya T,Nagano K,Yoshikawa T,Kamada H,Tsunoda S,Tsutsumi Y

doi

10.1016/j.cyto.2009.11.022

subject

Has Abstract

pub_date

2010-04-01 00:00:00

pages

75-83

issue

1

eissn

1043-4666

issn

1096-0023

pii

S1043-4666(09)00883-7

journal_volume

50

pub_type

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