Marianna Boncristiano

The Helicobacter pylori vacuolating toxin inhibits T cell activation by two independent mechanisms

Helicobacter pylori toxin, VacA, damages the gastric epithelium by erosion and loosening of tight junctions. Here we report that VacA also interferes with T cell activation by two different mechanisms. Formation of anion-specific channels by VacA prevents calcium influx from the extracellular milieu. The transcription factor NF-AT thus fails to translocate to the nucleus and activate key cytokine genes. A second, channel-independent mechanism involves activation of intracellular signaling through the mitogen-activated protein kinases MKK3/6 and p38 and the Rac-specific nucleotide exchange factor, Vav. As a consequence of aberrant Rac activation, disordered actin polymerization is stimulated. The resulting defects in T cell activation may help H. pylori to prevent an effective immune response leading to chronic colonization of its gastric niche.

Defective recruitment and activation of ZAP-70 in common variable immunodeficiency patients with T cell defects

We have previously identified a subset of common variable immunodeficiency (CVID) patients with defective T cell function associated with impaired activation of the TCR‐dependent tyrosine phosphorylation cascade. Here we have assessed the structural and functional integrity of the principal components involved in coupling the TCR/CD3 complex to intracellular tyrosine kinases in two of these patients. We show that ZAP‐70 fails to bind the signaling‐competent CD3ζ  tyrosine phosphorylation isoform and to become activated following TCR engagement, suggesting that defective recruitment of ZAP‐70 might underlie the TCR signaling dysfunction in these patients. Determination of the nucleotide sequences encoding the intracellular domains of the CD3/ζ  subunits and ZAP‐70 did not reveal any mutation. Furthermore, ZAP‐70 from these patients could interact in vitro with recombinant phospho‐ζ , ruling out genetic defects at the immunoreceptor tyrosine‐based activation motif/SH2 domain interface responsible for ZAP‐70 recruitment to the activated TCR. No defect was found in expression, activity or subcellular localization of Lck, which is thought to be primarily responsible for CD3ζ  phosphorylation. Hence, while the T cell defect in these CVID patients can be pinpointed to the interaction between ZAP‐70 and CD3ζ , the integrity in the components of the signaling machinery involved in this process suggests that additional components might be required for completion of this step.

Identification and Characterization of a Novel Nuclear Factor of Activated T-cells-1 Isoform Expressed in Mouse Brain

The nuclear factor of activated T-cells (NFAT) family transcription factors play a key role in the control of cytokine gene expression in T-cells. Although initially identified in T-cells, recent data have unveiled unanticipated roles for NFATs in the development, proliferation, and differentiation of other tissues. Here we report the identification, cDNA cloning, and functional characterization of a new isoform of NFAT1 highly expressed in mouse brain. This isoform, which we named NFAT1-D, is identical to NFAT1 throughout the N-terminal regulatory domain and the portion of the Rel domain which includes the minimal region required for specific binding to DNA and interaction with AP-1. The homology stops sharply upstream of the 3′-boundary of the Rel homology domain and is followed by a short unique C-terminal region. NFAT1-D was expressed at high levels in all brain districts and was found as a constitutively active transcription complex. Transfection of a NFAT/luciferase reporter in the neuronal cell line PC12, which also expresses NFAT1-D, showed that these cells expressed a constitutive NFAT activity that was enhanced after nerve growth factor-induced differentiation but was resistant to the immunosuppressant cyclosporin A. NFAT1-D was, however, inducibly activated in a cyclosporin A-sensitive manner when expressed in T-cells, suggesting that the activity of NFAT proteins might be controlled by their specific cellular context.

Dysregulation of the protein tyrosine kinase LCK in lymphoproliferative disorders and in other neoplasias.

Initially identified as a T-cell specific member of the Src family of protein tyrosine kinases, Lck has become the object of intensive investigations which have revealed a key role for this kinase in the central processes controlling T-cell development, activation, proliferation and survival. Experimental evidence of the oncogenic potential of Lck, together with the identification of defects in the regulation of Lck expression or activity in T-cell leukemias, suggests that dysregulation of Lck might play a role in neoplastic transformation. Here we review the data documenting a potential role for this kinase in the initiation and maintenance of the transformed state in human cancers.

Molecular mechanisms underlying suppression of lymphocyte responses by nonsteroidal antiinflammatory drugs.

Abstract:Initially identified and further developed as inhibitors of cyclooxygenases, nonsteroidal antiinflammatory drugs (NSAIDs) have been more recently shown to bind to and act as agonists of the peroxisome proliferator-activated receptor family of transcription factors. Here we summarize the current knowledge on the functions of the principal targets of NSAIDs and review their role in T and B lymphocytes, with a focus on the molecular mechanisms underlying the effects of NSAIDs on lymphocyte development, activation, differentiation and death.