MUTATION Mutations in have so far been reported in immunodeficient patients from only one consanguineous family

MUTATION Mutations in have so far been reported in immunodeficient patients from only one consanguineous family. review describes the immunological and non-immunological phenotypes of patients with defects in SOCE and CRAC channel function and discusses them in the context of similar immunodeficiency diseases and animal models of ORAI1 and STIM1 function. as the pore forming subunit of the CRAC channel and stromal interaction molecule 1 (or [22] and the drosophila gene MUTATIONS Autosomal recessive mutations in the gene have been reported in six patients from three unrelated families. All patients lack store-operated Ca2+ entry in T cells and other cell types and suffer from severe immunodeficiency with recurrent and opportunistic infections. They have normal lymphocyte numbers and subsets but T cell activation is severely compromised with strongly impaired proliferative responses in vitro. ORAI1-R91W missense mutation Two patients were born to consanguineous parents of Turkish origin (Figure 4A). Polyclonal T cell lines established from the patients lack SOCE in reponse to TCR stimulation and thapsigargin, an inhibitor of the sarcoplasmic/endoplasmic reticulum Ca2+ ATPase (SERCA), or ionomycin, both of which passively deplete ER Ca2+ stores and activate the CRAC channel [9]. A CRAC channel defect was established in these patients as ICRAC was undetectable ruling out indirect effects on SOCE as the cause of disease [9]. Both patients are homozygous for a missense mutation in that results in the substitution of a highly conserved arginine residue at the beginning of its first transmembrane domain with tryptophane (and in SBI-0206965 immunodeficient patients lacking Ca2+ influxA, ORAI1 is a plasma membrane Ca2+ channel with four transmembrane domains and intracellular N- and C-termini (for details see text). Mutations: (i) An ArgTrp (R91W) single amino acid substitution in ORAI1 at the beginning of the first transmembrane (TM1) domain results in expression of a nonfunctional protein [17]. (ii) A frameshift (fs) nonsense mutation in the first exon of ORAI1 (A88SfsX25) results in premature termination, nonsense mediated decay of ORAI1 mRNA and abolished ORAI1 protein expression [33]. (iii) Two independent single amino acid substitutions, A103E and L194P, in TM1 and TM3, respectively interfere with ORAI1 protein expression SBI-0206965 [33]. B, STIM1 is a single-pass transmembrane (TM) protein localized in the ER featuring an EF hand domain (EFh), sterile alpha motif (SAM), coiled-coil (CC), serin/proline (S/P) and lysine (K) rich domains (for details see text). Mutation: A frameshift nonsense mutation in exon 3 of STIM1 (E128RfsX9) results in premature termination, nonsense mediated mRNA decay of STIM1 and abolished STIM1 protein expression [19]. ORAI1 expression is abolished in patients from two families unrelated to the family described above. An which results in a frame shift and premature termination at position 112 (resulting in single amino acid substitutions in the first (A103E) and third (L194P) transmembrane domain of ORAI1 (Figure 5A) [33]. ORAI1 protein was not detectable in the patients fibroblasts or in HEK293 cells ectopically expressing the ORAI1 mutants whereas ORAI1 mRNA levels were normal. Both mutations interfere with protein expression, presumably by destablizing the transmembrane domains in which they are located. The patients parents are heterozygous for either one of the two mutations, but the presence of one wild-type allele results in normal SOCE and prevents immunodeficiency. 2.3. CLINICAL PHENOTYPE Immunodeficiency Immunodeficiency in ORAI1 deficient patients is characterized by recurrent severe infections with viral, bacterial, mycobacterial and fungal pathogens causing pneumonia, meningitis, enteritis, gastrointestinal candidiasis and sepsis (Table 1)[10; 11; 18; 33; 36]. Antibiotic and intravenous immunoglobulin therapy are only partially effective in controlling infections in the ORAI1 deficient patients. One patient with and absent SBI-0206965 cutaneous delayed-type hypersensitivity reactions [10; 11; 18; 33; 36]. Interestingly, T cells from patients lacking ORAI1 expression proliferated normally in response to stimulation with PMA Rabbit Polyclonal to Keratin 20 and ionomycin whereas T cells from patients expressing a non-functional form of ORAI1 (R91W) did not. The difference may be due to a potential compensatory effect of ORAI2 or ORAI3 that occurs in the absence of ORAI1, indicating that the R91W mutation may exert a SBI-0206965 moderate inhibitory effect on SOCE. This finding is consistent with partially impaired SOCE in T cells of heterozygous carriers of the R91W mutation in contrast to normal SOCE in T cells from individuals with monoallelic expression of wild-type ORAI1 [33]. It is important to note, however, that heterozygous carriers of the various mutations in (and presented with an inability to sweat and recurrent fever episodes due to impaired thermoregulation, especially in the summer, in the majority of ORAI1 deficient patients [33]. In the absence of available skin biopsies, it remains unclear whether eccrine sweat glands do not develop or are non-functional in these.