Analysis of the oligomeric requirement for signaling by CD40 using soluble multimeric forms of its ligand, CD154

Haswell, Linsey E.; Glennie, Martin J.; Al‐Shamkhani, Aymen

doi:10.1002/1521-4141(2001010)31:10<3094::aid-immu3094>3.0.co;2-f

Cited by 94 publications

(66 citation statements)

References 35 publications

(42 reference statements)

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“…CD40/CD40 Homodimer Formation Is an Absolute Requirement for B7.2 Expression-To further explore the biological outcomes of CD40/CD40 homodimer engagement, we analyzed the transcription of B7.2, a strong T-cell co-stimulatory molecule regulated following CD40 ligation on B cells (34,35). Our results indicate that CD40 ligation by m-CD154 enhanced B7.2 transcription on BJAB B cells after a 2-h co-incubation period (Fig.…”

Section: Cd40/cd40 Homodimer Formation Is Required For Cd40-induced Pmentioning

confidence: 98%

CD40/CD40 Homodimers Are Required for CD40-induced Phosphatidylinositol 3-Kinase-dependent Expression of B7.2 by Human B Lymphocytes

Reyes‐Moreno

Girouard

Lapointe

et al. 2004

Journal of Biological Chemistry

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Preformed CD40/CD40 homodimers were initially observed on human Burkitt lymphoma cell lines, normal B cells, and transitional bladder carcinoma cell lines. However, the nature and the biological relevance of these homodimers have not yet been investigated. In the present study, we demonstrated that Epstein-Barr virus-transformed B cells and CD40-transfected HEK 293 cells constitutively expressed disulfide-linked CD40/ CD40 homodimers at low levels. Oligomerization of CD40 leads to a rapid and significant increase in the disulfide-linked CD40/CD40 homodimer formation, a response that could be prevented using a thiol-alkylating agent. Formation of CD40/CD40 homodimers was found to be absolutely required for CD40-mediated activation of phosphatidylinositol 3-kinase, which, in turn regulated B7.2 expression. In contrast, CD40 monomers provided the minimal signal emerging from CD40, activating p38 MAP kinase and inducing homotypic B cell adhesion. CD40/CD40 homodimer formation was totally independent of TRAF1/2/3/5 associations with the threonine at position 254 in the cytoplasmic tail of the CD40 molecules. This finding may be vital to better understanding the molecular mechanisms that govern cell signaling triggered by CD40/CD154 interactions.The CD40 molecule is a 45-50-kDa type I phosphorylated glycoprotein that belongs to the tumor necrosis factor receptor superfamily and was initially considered to be a growth factor receptor on B cells (1). The interaction of CD40 with its natural ligand, CD154, is a crucial step in T cell-dependent B cell activation and differentiation as demonstrated in patients suffering from the X-linked hyper-IgM syndrome (2) and then confirmed in CD154 (3) and CD40 knock-out mice (4). Although CD40 has no kinase domain, its ligation induces the activation of protein tyrosine kinases (including lyn, fin, and syk) (5), the adaptor protein jak3 (6), the phosphatidylinositol-3 kinase (PI 3-kinase) 1 (7), the phospholipase C␥2 (7), and the mitogenactivated protein (MAP) kinases p38, c-Jun N-terminal kinase/ stress-activated protein kinase (JNK/SAPK), and extracellular signal-regulated kinase 1 or 2 (ERK1/2) (8 -10). Most of these early events are mediated by the association of CD40 via its cytoplasmic domain with several members of the tumor necrosis factor receptor-associated factor (TRAF) family (11-17).The cellular events triggered in CD40-positive cells following CD40/CD154 interactions are not restricted to B cells only but can also be triggered in dendritic cells, monocytes, epithelial cells, endothelial cells, and fibroblasts (1). Thus, CD40 ligation mediates a broad variety of immune and inflammatory responses (1, 18). Abnormal CD40 signaling seems to be directly associated with the pathogenesis of chronic inflammatory diseases such as autoimmune diseases (19), neurodegenerative disorders (20), graft-versus-host diseases (21), cardiovascular diseases (22), and cancer (23). Consequently, elucidation of the molecular mechanisms that govern CD40-mediated signal transduction is critical n...

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Section: Cd40/cd40 Homodimer Formation Is Required For Cd40-induced Pmentioning

confidence: 98%

CD40/CD40 Homodimers Are Required for CD40-induced Phosphatidylinositol 3-Kinase-dependent Expression of B7.2 by Human B Lymphocytes

Reyes‐Moreno

Girouard

Lapointe

et al. 2004

Journal of Biological Chemistry

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“…The question of whether it will be more effective to use a human (or humanized) IgG anti-CD40 and run the risk of generating anti-Id responses or to use a soluble recombinant ligand, such as a CD154 trimer, remains to be seen. In this respect recently work has shown that a dodecamer of CD154 is far more active than trimeric CD154 at activating cells via CD40 (42).…”

Section: Figurementioning

confidence: 99%

T Cell Immunity to Lymphoma Following Treatment with Anti-CD40 Monoclonal Antibody

Tutt

O’Brien

Hussain

et al. 2002

The Journal of Immunology

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In this study we demonstrate that treatment with anti-CD40 mAb eradicates a range of mouse lymphomas (BCL1, A31, A20, and EL4), but only when used against i.v. tumor doses in excess of 107 cells. Only partial protection was seen against smaller tumor loads. We saw no evidence that anti-CD40 mAb changed the phenotype of the lymphomas or inhibited their growth in the initial period following treatment, but it did result in a rapid expansion of cytotoxic CD8+ cells that was able to clear the neoplastic disease and provide long-term protection against tumor rechallenge. The CTL responses were blocked by mAb against a range of coreceptors and cytokines, including CD8, B7-1, B7-2, LFA-1, and IFN-γ, but not CD4 or CTLA-4, indicating the presence of a conventional cellular Th1 response. Furthermore, we found evidence of cross-recognition between lymphomas (BCL1 and A20) as measured by cytotoxicity and IFN-γ responses in vitro and using tumor rechallenge experiments, suggesting common target Ags. Finally, although anti-CD40 was shown to stimulate NK cell killing, we could find no role for these cells in controlling tumor growth. These data underline the ability of anti-CD40 mAb to potentiate CTL responses and the potency of cellular immunity in eradicating large quantities of syngeneic tumor.

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“…For example, the soluble forms of TNF, CD95L, TRAIL, and CD40L interact with TNFR2, CD95, TRAILR2, and CD40, respectively, but do not or only poorly activate signaling by these receptors. [3][4][5][6] Notably, inactive or poorly active soluble TNF ligands can be converted into highly active molecules by artificially increasing their avidity. For example, soluble flag-tagged variants of TNF, CD95L, TRAIL, and CD40L stimulate robust signaling by TNFR2, CD95, TRAILR2, and CD40, respectively, provided they were cross-linked with the Flag-specific mAb M2.…”

mentioning

confidence: 99%

“…Likewise, hexameric and dodecameric fusion proteins of soluble CD95L and soluble CD40L as well as non-specifically aggregated preparations of TNF ligands produced in E. coli display high activity. [6][7][8] The structural hallmark of the ligands of the TNF family is the carboxy-terminal 'TNF homology domain', which is part of both the transmembrane and soluble forms of TNF ligands. [1][2] The TNF homology domains (THDs) of the various TNF ligands are composed of a framework of aromatic and hydrophobic residues that adopt an almost identical tertiary fold and cause self association into trimers.…”

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confidence: 99%

Enforced covalent trimerization increases the activity of the TNF ligand family members TRAIL and CD95L

et al. 2007

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Variants of human TRAIL (hTRAIL) and human CD95L (hCD95L), encompassing the TNF homology domain (THD), interact with the corresponding receptors and stimulate CD95 and TRAILR2 signaling after cross-linking. The murine counterparts (mTRAIL, mCD95L) showed no or only low receptor binding and were inactive/poorly active after cross-linking. The stalk region preceding the THD of mCD95L conferred secondary aggregation and restored CD95 activation in the absence of cross-linking. A corresponding variant of mTRAIL, however, was still not able to activate TRAIL death receptors, but gained good activity after cross-linking. Notably, disulfide-bonded fusion proteins of the THD of mTRAIL and mCD95L with a subdomain of the tenascin-C (TNC) oligomerization domain, which still assembled into trimers, efficiently interacted with their cognate cellular receptors and robustly stimulated CD95 and TRAILR2 signaling after secondary cross-linking. Introduction of the TNC domain also further enhanced the activity of THD encompassing variants of hTRAIL and hCD95L. Thus, spatial fixation of the N-terminus of the THD appears necessary in some TNF ligands to ensure proper receptor binding. This points to yet unanticipated functions of the stalk and/or transmembrane region of TNF ligands for the functionality of these molecules and offers a broadly applicable option to generate recombinant soluble ligands of the TNF family with superior activity. Ligands of the tumor necrosis factor (TNF) family fulfill crucial roles in the immune system, but have also been implicated in the development of epithelial and endothelial structures. 1 TNF family ligands are primarily expressed as trimeric type II transmembrane proteins and are often processed into soluble variants that are also organized as trimers. 1,2 Although shedding of some TNF ligands does not interfere with their capability to activate their corresponding receptors and might be even important for their physiological function, other TNF ligands become inactivated by proteolytic processing. 2 Soluble TNF ligands that are not or only poorly active still interact with their cognate receptors. For example, the soluble forms of TNF, CD95L, TRAIL, and CD40L interact with TNFR2, CD95, TRAILR2, and CD40, respectively, but do not or only poorly activate signaling by these receptors. [3][4][5][6] Notably, inactive or poorly active soluble TNF ligands can be converted into highly active molecules by artificially increasing their avidity. For example, soluble flag-tagged variants of TNF, CD95L, TRAIL, and CD40L stimulate robust signaling by TNFR2, CD95, TRAILR2, and CD40, respectively, provided they were cross-linked with the Flag-specific mAb M2. Likewise, hexameric and dodecameric fusion proteins of soluble CD95L and soluble CD40L as well as non-specifically aggregated preparations of TNF ligands produced in E. coli display high activity. [6][7][8] The structural hallmark of the ligands of the TNF family is the carboxy-terminal 'TNF homology domain', which is part of both the transmembrane and s...

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Analysis of the oligomeric requirement for signaling by CD40 using soluble multimeric forms of its ligand, CD154

Cited by 94 publications

References 35 publications

CD40/CD40 Homodimers Are Required for CD40-induced Phosphatidylinositol 3-Kinase-dependent Expression of B7.2 by Human B Lymphocytes

CD40/CD40 Homodimers Are Required for CD40-induced Phosphatidylinositol 3-Kinase-dependent Expression of B7.2 by Human B Lymphocytes

T Cell Immunity to Lymphoma Following Treatment with Anti-CD40 Monoclonal Antibody

Enforced covalent trimerization increases the activity of the TNF ligand family members TRAIL and CD95L

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