Costimulatory B7 molecules (B7-1 or CD80;and B7-2 or CD86) are known to bind to T cell costimulatory receptors CD28 and CTLA4. Engagement of CD28 is know to transduce signals in T cells that play a critical role in T cell activation. TCR mediated activation of T cells to proliferation and IL2 secretion was enhanced in mice deficient in the adapter molecule cbl-b, suggesting a negative regulatory role for cbl-b under these conditions. Cbl-b deletion in fact reversed the inability of CD28-deficient mice to generate T cell-dependent Ig class switched primary and secondary antibody responses. The observation that cbl-b inactivation can enhance T cell responses to TCR signaling, and can bypass requirements for CD28 costimulation has been translated to studies of tumor rejection. Inactivation of cbl-b enhances the ability of mice to reject tumors that do not express costimulatory ligands (CD80 or CD86), and that grow progressively in wild type mice. Cbl-b inactivation also dramatically inhibits the incidence of thymic lymphomas in ATM-deficient mice. These findings identify a novel strategy for enhancing T cell-mediated tumor immunotherapy by modulating regulatory influences on T cell activation. Translational efforts are in progress using high throughput screening to identfy molecules that are capable of reproducing the effects of cbl-b inactivation, allowing human T lymphocytes to respond to T cell receptor (TCR) stimuli in the absence of otherwise required CD28-mediate cosimulation. The relationship between members of the cbl family and other critical adapter molecules has been studied. Experiments have indicated that inactivation of c-cbl (but not cbl-b) completely reverses the lethal phenotype caused by SLP-76 deficiency. In addition, c-cbl inactivation partially reverses the defect in T cell development caused by deficiency in SLP-76, LAT,or Vav1. These findings indicate an unanticipated SLP-76 (and LAT) independent signaling pathway that is facilitated by c-cbl inactivation. The biochemical basis for these effects has been studied using SLP-76 mutants to dissect the structure-function relationships in this pathway. C-cbl specifically rescues the T cell developmental defect resulting from SLP-76 mutations that disrupt tyrosines involved in binding to Vav and Itk. Subsequent analysis has further indicated that c-cbl inactivation rescues the developmental defect in Vav1-deficient mice, further characterizing a SLP-76/LAT/Vav1 pathway for T cell development. In contrast, c-cbl inactivation fails to overcome the T cell developmental defects that occur un lck-deficient mice. Thus, the pathways unmasked by c-cbl deficiency appear to require lck. The molecular components of this pathway are currently under study. The src family kinase lck plays a critical role in T lymphocyte development and activation. Expression of lck is regulated by two promoters, termed distal and proximal promoters, in both mice and humans. Although the existence of these promoters was desribed 20 years ago, their function in developing and mature T cels has not been analyzed. To address this question, we have generated mutant BAC transgenes by deletion of either proximal or distal lck promoter. Studies of these mice have demonstrated selective expression of lck at preferential and different stages of development when driven by one or the other of its two promoters. Corresponding slective stages of T cell development and differentiation have been shown to be dependent upon promoter-specific lck expression. In addition to conventional downstream events in TCR or cytokine signaling, we have identified novel effects of the tumor suppressor p53 in regulating responses to these stimuli. We have observed differential induction of p53 by distinct stimuli and consequent effects of p53 on functional responses of T cells. The molecular mechanisms mediating these effects are being studied.
|Williams, Joy A; Zhang, Jingjing; Jeon, Hyein et al. (2014) Thymic medullary epithelium and thymocyte self-tolerance require cooperation between CD28-CD80/86 and CD40-CD40L costimulatory pathways. J Immunol 192:630-40|
|Watanabe, Masashi; Moon, Kyung Duk; Vacchio, Melanie S et al. (2014) Downmodulation of tumor suppressor p53 by T cell receptor signaling is critical for antigen-specific CD4(+) T cell responses. Immunity 40:681-91|
|Fujihara, Chiharu; Williams, Joy A; Watanabe, Masashi et al. (2014) T cell-B cell thymic cross-talk: maintenance and function of thymic B cells requires cognate CD40-CD40 ligand interaction. J Immunol 193:5534-44|