PMID- 19596991
OWN - NLM
STAT- MEDLINE
DCOM- 20090806
LR  - 20210103
IS  - 1550-6606 (Electronic)
IS  - 0022-1767 (Linking)
VI  - 183
IP  - 3
DP  - 2009 Aug 1
TI  - Trimer stabilization, oligomerization, and antibody-mediated cell surface 
      immobilization improve the activity of soluble trimers of CD27L, CD40L, 41BBL, 
      and glucocorticoid-induced TNF receptor ligand.
PG  - 1851-61
LID - 10.4049/jimmunol.0802597 [doi]
AB  - For many ligands of the TNF family, trimer stability and oligomerization status 
      are crucial determinants of receptor activation. However, for the 
      immunostimulatory ligands CD27L, CD40L, 41BBL, and glucocorticoid-induced TNF 
      receptor ligand (GITRL) detailed information regarding these requirements is 
      lacking. Here, we comprehensively evaluated the effect of trimer stability and 
      oligomerization on receptor activation by these ligands. Treatment with soluble 
      Flag-tagged CD27L, 41BBL, and GITRL minimally activated receptor signaling, while 
      Flag-CD40L was highly active. Oligomerization with anti-Flag Abs further enhanced 
      the specific activity of Flag-CD40L 10-fold and of Flag-41BBL more than 200-fold, 
      but it failed to activate Flag-CD27L and Flag-GITRL. We next investigated the 
      relevance of trimer stability by introducing the tenascin-C (TNC) trimerization 
      domain, yielding stabilized Flag-TNC-ligand trimers. Oligomerization with 
      anti-Flag Ab potently activated signaling by Flag-TNC-CD27L and Flag-TNC-GITRL 
      and, albeit to a lesser extent, Flag-TNC-CD40L and Flag-TNC-41BBL. Forced 
      hexamerization, by introducing an Ig Fc domain, revealed that hexameric 
      derivatives of Flag-TNC-41BBL, Flag-CD40L, and Flag-TNC-GITRL all activate 
      receptor signaling with high efficiency, whereas hexameric Flag-CD27L variant 
      left inactive. Finally, we attempted to selectively activate receptor signaling 
      on targeted cells, by using Ab fragment (single-chain fragment variable region, 
      scFv)-ligand fusion proteins, an approach previously applied to other TNF 
      ligands. Target cell surface Ag-selective activation was achieved for scFv-41BBL, 
      scFv-CD40L, and scFv-GITRL, although the latter two displayed already significant 
      activity toward Ag-negative cells. In conclusion, our data establish that 
      trimeric CD40L is active, 41BBL requires hexamerization, GITRL requires trimer 
      stabilization, and CD27L requires trimer stabilization and oligomerization. 
      Furthermore, surface immobilization might be exploited to gain locally enhanced 
      ligand activity.
FAU - Wyzgol, Agnes
AU  - Wyzgol A
AD  - Division of Molecular Internal Medicine, Department of Internal Medicine II, 
      University Hospital Wuerzburg, Wuerzburg, Germany.
FAU - Muller, Nicole
AU  - Muller N
FAU - Fick, Andrea
AU  - Fick A
FAU - Munkel, Sabine
AU  - Munkel S
FAU - Grigoleit, Goetz Ulrich
AU  - Grigoleit GU
FAU - Pfizenmaier, Klaus
AU  - Pfizenmaier K
FAU - Wajant, Harald
AU  - Wajant H
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20090713
PL  - United States
TA  - J Immunol
JT  - Journal of immunology (Baltimore, Md. : 1950)
JID - 2985117R
RN  - 0 (4-1BB Ligand)
RN  - 0 (Antibodies)
RN  - 0 (CD27 Ligand)
RN  - 0 (Receptors, Tumor Necrosis Factor)
RN  - 0 (Tenascin)
RN  - 0 (Tnfsf18 protein, mouse)
RN  - 0 (Tumor Necrosis Factors)
RN  - 147205-72-9 (CD40 Ligand)
SB  - IM
MH  - 4-1BB Ligand/immunology/metabolism
MH  - Animals
MH  - Antibodies/metabolism
MH  - CD27 Ligand/immunology/metabolism
MH  - CD40 Ligand/*immunology/*metabolism
MH  - Mice
MH  - *Protein Multimerization
MH  - Protein Stability
MH  - Receptors, Tumor Necrosis Factor/immunology/*metabolism
MH  - Solubility
MH  - Tenascin/pharmacology
MH  - Tumor Necrosis Factors/immunology/*metabolism
EDAT- 2009/07/15 09:00
MHDA- 2009/08/07 09:00
CRDT- 2009/07/15 09:00
PHST- 2009/07/15 09:00 [entrez]
PHST- 2009/07/15 09:00 [pubmed]
PHST- 2009/08/07 09:00 [medline]
AID - jimmunol.0802597 [pii]
AID - 10.4049/jimmunol.0802597 [doi]
PST - ppublish
SO  - J Immunol. 2009 Aug 1;183(3):1851-61. doi: 10.4049/jimmunol.0802597. Epub 2009 
      Jul 13.