Antiviral and regulatory T cell immunity in a patient with stromal interaction molecule 1 deficiency.

Stromal interaction molecule 1 (STIM1) deficiency is a rare genetic disorder of store-operated calcium entry, associated with a complex syndrome including immunodeficiency and immune dysregulation. The link from the molecular defect to these clinical manifestations is incompletely understood. We report two patients with a homozygous R429C point mutation in STIM1 completely abolishing store-operated calcium entry in T cells. Immunological analysis of one patient revealed that despite the expected defect of T cell proliferation and cytokine production in vitro, significant antiviral T cell populations were generated in vivo. These T cells proliferated in response to viral Ags and showed normal antiviral cytotoxicity. However, antiviral immunity was insufficient to prevent chronic CMV and EBV infections with a possible contribution of impaired NK cell function and a lack of NKT cells. Furthermore, autoimmune cytopenia, eczema, and intermittent diarrhea suggested impaired immune regulation. FOXP3-positive regulatory T (Treg) cells were present but showed an abnormal phenotype. The suppressive function of STIM1-deficient Treg cells in vitro, however, was normal. Given these partial defects in cytotoxic and Treg cell function, impairment of other immune cell populations probably contributes more to the pathogenesis of immunodeficiency and autoimmunity in STIM1 deficiency than previously appreciated.

ORAI1-mediated calcium influx is required for human cytotoxic lymphocyte degranulation and target cell lysis.

Lymphocytes mediate cytotoxicity by polarized release of the contents of cytotoxic granules toward their target cells. Here, we have studied the role of the calcium release-activated calcium channel ORAI1 in human lymphocyte cytotoxicity. Natural killer (NK) cells obtained from an ORAI1-deficient patient displayed defective store-operated Ca(2+) entry (SOCE) and severely defective cytotoxic granule exocytosis leading to impaired target cell lysis. Similar findings were obtained using NK cells from a stromal interaction molecule 1-deficient patient. The defect occurred at a late stage of the signaling process, because activation of leukocyte functional antigen (LFA)-1 and cytotoxic granule polarization were not impaired. Moreover, pharmacological inhibition of SOCE interfered with degranulation and target cell lysis by freshly isolated NK cells and CD8(+) effector T cells from healthy donors. In addition to effects on lymphocyte cytotoxicity, synthesis of the chemokine macrophage inflammatory protein-1ß and the cytokines TNF-α and IFN-γ on target cell recognition was impaired in ORAI1-deficient NK cells, as previously described for T cells. By contrast, NK cell cytokine production induced by combinations of IL-12, IL-15, and IL-18 was not impaired by ORAI1 deficiency. Taken together, these results identify a critical role for ORAI1-mediated Ca(2+) influx in granule exocytosis for lymphocyte cytotoxicity as well as for cytokine production induced by target cell recognition.

Differential expression of four protocadherin alpha and gamma clusters in the developing and adult zebrafish: DrPcdh2gamma but not DrPcdh1gamma is expressed in neuronal precursor cells, ependymal cells and non-neural epithelia.

Protocadherins are cadherin-like molecules with adhesive and signaling functions, in particular, during neuronal development. Large protocadherin (Pcdh) gene clusters are present in the genome of vertebrates. In the zebrafish, two Pcdh clusters are found on chromosomes 10 (DrPcdh1) and 14 (DrPcdh2), each divided into subclusters of DrPcdhalpha and DrPcdhgamma family genes. In total, about 100 different DrPcdh molecules are predicted. We have analyzed the expression of the four DrPcdh subclusters and find that DrPcdh transcripts are upregulated in the developing zebrafish nervous system. In the adult fish brain, all four DrPcdh clusters are expressed in differentiated neurons, in particular, in the thalamic nuclei, tectum, and cerebellum. We show that expression patterns grossly overlap for each cluster but with regional differences and variations in strength of expression. Strikingly, the DrPcdh2gamma cluster, distinct from the three other clusters, is also expressed in neuronal precursor cells and ependymal cells of the embryonic and adult nervous system, as well as in specific non-neuronal epithelia. Antibodies to a conserved motif in the constant region of DrPcdh2gamma stain fiber tracts and neuropil of the zebrafish brain and cell-cell junctions in epithelia. Our results indicate that multiple DrPcdhs of the different clusters are expressed in differentiated zebrafish neurons, suggesting evolutionarily conserved functions of protocadherin clusters in cell adhesion and signaling. In addition, DrPcdh2gamma may exert more specific roles in neuronal precursor and non-neural epithelial cells, which have not yet been described for mammalian Pcdhgamma. Thus, our findings in zebrafish open new perspectives to examine these functions in other vertebrate model organisms.