CD40, a member of the tumor necrosis factor (TNF) receptor family, plays an essential role in T cell–dependent immune responses. Because CD40 is widely expressed on the surface of tumor cells in various B cell malignancies, deregulated CD40 signaling has been suggested to contribute to lymphomagenesis. In this study, we show that B cell-specific expression of a constitutively active CD40 receptor, in the form of a latent membrane protein 1 (LMP1)/CD40 chimeric protein, promoted an increase in the number of follicular and marginal zone B cells in secondary lymphoid organs in transgenic mice. The B cells displayed an activated phenotype, prolonged survival and increased proliferation, but were significantly impaired in T cell-dependent immune responses. Constitutive CD40 signaling in B cells induced selective and constitutive activation of the noncanonical NF-κB pathway and the mitogen-activated protein kinases Jnk and extracellular signal–regulated kinase. LMP1/CD40-expressing mice older than 12 mo developed B cell lymphomas of mono- or oligoclonal origin at high incidence, thus showing that the interplay of the signaling pathways induced by constitutive CD40 signaling is sufficient to initiate a tumorigenic process, ultimately leading to the development of B cell lymphomas.

No abstract

CD40, a member of the TNF receptor family, is expressed on all mature B cells and on most B-cell lymphomas. Recently, we have shown that constitutive activation of CD40 signaling in B cells induced by a fusion protein consisting of the transmembrane part of the Epstein-Barr viral latent membrane protein 1 (LMP1) and the cytoplasmic part of CD40 (LMP1/CD40) drives B-cell lymphoma development in transgenic mice. Because LMP1/ CD40-expressing B cells showed an upregulation of CD19, we investigated CD19's function in CD40-driven B-cell expansion and lymphomagenesis. Here, we demonstrate that ablation of CD19 in LMP1/CD40 transgenic mice resulted in a severe loss and reduced lifespan of mature B cells and completely abrogated development of B-cell lymphoma. CD19 is localized to lipid rafts and constitutively activated by the LMP1/CD40 fusion protein in B cells. We provide evidence that the improved survival and malignant transformation of LMP1/CD40-expressing B cells are dependent on activation of the MAPK Erk that is mediated through CD19 in a PI3K-dependent manner. Our data suggest that constitutively active CD40 is dependent on CD19 to transmit survival and proliferation signals. Moreover, we detected a similarly functioning prosurvival pathway involving phosphorylated CD19 and PI3K-dependent Erk phosphorylation in human diffuse large B-cell lymphoma cell lines. Our data provide evidence that CD19 plays an important role in transmitting survival and proliferation signals downstream of CD40 and therefore might be an interesting therapeutic target for the treatment of lymphoma undergoing chronic CD40 signaling. Cancer Res; 74(16); 4318-28. Ó2014 AACR.

<p>PDF file - 364K, Suppl. Figure 1. BM B cell development is comparable in LMP1/CD40//CD19+/- and LMP1/CD40//CD19-/- mice. Suppl. Figure 2. CD40 employs CD19 to mediate survival signals. Suppl. Figure 3. Loss of CD19 abrogates lymphoma development in LMP1/CD40 mice. Suppl. Figure 4A. Stimulation of B cells with anti-IgM in the presence of Dasatinib. Suppl. Figure 4B. Titration of the PI3K inhibitor LY294004 Suppl. Figure 5A. Sorafenib does not affect Erk phosphorylation in LMP1/CD40+ B cells. Suppl. Figure 5B. CD19 expression on different DLBCL and HL cell lines. Suppl. Figure 6A. CD40 expression in different DLBCL cell lines. Suppl. Figure 6B. CD19 phosphorylation after stimulation with a CD40 ligand.</p>

<p>PDF file - 364K, Suppl. Figure 1. BM B cell development is comparable in LMP1/CD40//CD19+/- and LMP1/CD40//CD19-/- mice. Suppl. Figure 2. CD40 employs CD19 to mediate survival signals. Suppl. Figure 3. Loss of CD19 abrogates lymphoma development in LMP1/CD40 mice. Suppl. Figure 4A. Stimulation of B cells with anti-IgM in the presence of Dasatinib. Suppl. Figure 4B. Titration of the PI3K inhibitor LY294004 Suppl. Figure 5A. Sorafenib does not affect Erk phosphorylation in LMP1/CD40+ B cells. Suppl. Figure 5B. CD19 expression on different DLBCL and HL cell lines. Suppl. Figure 6A. CD40 expression in different DLBCL cell lines. Suppl. Figure 6B. CD19 phosphorylation after stimulation with a CD40 ligand.</p>