Transmembrane TNF-dependent uptake of anti-TNF antibodies.
TNF-α (TNF), a pro-inflammatory cytokine is synthesized as a 26 kDa protein, anchors in the plasma membrane as transmembrane TNF (TmTNF), and is subjected to proteolysis by the TNF-α converting enzyme (TACE) to release the 15 kDa form of soluble TNF (sTNF). TmTNF and sTNF interact with 2 distinct receptors, TNF-R1 (p55) and TNF-R2 (p75), to mediate the multiple biologic effects of TNF described to date. Several anti-TNF biologics that bind to both forms of TNF and block their interactions with the TNF receptors are now approved for the treatment of a variety of immune-mediated diseases. Several reports suggest that binding of anti-TNFs to TmTNF delivers an outside-to-inside 'reverse' signal that may also contribute to the efficacy of anti-TNFs. Some patients, however, develop anti-TNF drug antibody responses (ADA or immunogenicity). Here, we demonstrate biochemically that TmTNF is transiently expressed on the surface of lipopolysaccharide-stimulated primary human monocytes, macrophages, and monocyte-derived dendritic cells (DCs) and expression of TmTNF on the cell surface is enhanced following treatment of cells with TAPI-2, a TACE inhibitor. Importantly, binding of anti-TNFs to TmTNF on DCs results in rapid internalization of the anti-TNF/TmTNF complex first into early endosomes and then lysosomes. The internalized anti-TNF is processed and anti-TNF peptides can be eluted from the surface of DCs. Finally, tetanus toxin peptides fused to anti-TNFs are presented by DCs to initiate T cell recall proliferation response. Collectively, these observations may provide new insights into understanding the biology of TmTNF, mode of action of anti-TNFs, biology of ADA response to anti-TNFs, and may help with the design of the next generation of anti-TNFs.
a AbbVie Bioresearch Center , Worcester , MA , USA.
Arun
A
Deora
a AbbVie Bioresearch Center , Worcester , MA , USA.
Subramanya
S
Hegde
a AbbVie Bioresearch Center , Worcester , MA , USA.
Jacqueline
J
Lee
a AbbVie Bioresearch Center , Worcester , MA , USA.
Chee-Ho
CH
Choi
a AbbVie Bioresearch Center , Worcester , MA , USA.
Qing
Q
Chang
a AbbVie Bioresearch Center , Worcester , MA , USA.
Cheryl
C
Lee
a AbbVie Bioresearch Center , Worcester , MA , USA.
Lucia
L
Eaton
b AbbVie Inc , North Chicago , IL , USA.
Hua
H
Tang
a AbbVie Bioresearch Center , Worcester , MA , USA.
Dongdong
D
Wang
a AbbVie Bioresearch Center , Worcester , MA , USA.
David
D
Lee
a AbbVie Bioresearch Center , Worcester , MA , USA.
Mark
M
Michalak
a AbbVie Bioresearch Center , Worcester , MA , USA.
Medha
M
Tomlinson
a AbbVie Bioresearch Center , Worcester , MA , USA.
Qingfeng
Q
Tao
a AbbVie Bioresearch Center , Worcester , MA , USA.
Nidhi
N
Gaur
a AbbVie Bioresearch Center , Worcester , MA , USA.
Bohdan
B
Harvey
b AbbVie Inc , North Chicago , IL , USA.
Shaun
S
McLoughlin
a AbbVie Bioresearch Center , Worcester , MA , USA.
Boris
B
Labkovsky
a AbbVie Bioresearch Center , Worcester , MA , USA.
Tariq
T
Ghayur
D000818:Animals; D000906:Antibodies / Q000378:metabolism / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism*; D003713:Dendritic Cells / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D008264:Macrophages / Q000378:metabolism; D051379:Mice; D009000:Monocytes / Q000378:metabolism; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000378:metabolism*; D047889:Receptors, Tumor Necrosis Factor, Type II / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors / Q000378:metabolism*
D000906:Antibodies; D047888:Receptors, Tumor Necrosis Factor, Type I; D047889:Receptors, Tumor Necrosis Factor, Type II; D014409:Tumor Necrosis Factor-alpha
Anti-drug antibodies (ADA); TmTNF reverse signaling; Transmembrane TNF; anti-TNFs; clathrin; endocytosis; membrane TNF; tetanus toxin