Genetic basis of chronic mucocutaneous candidiasis disease in humans

• Main Author: Lévy, Romain
• Other Authors: Sorbonne Paris Cité
• Language: en
• Published: 2017
• Subjects: Génétique humaine; Séquençage de l’exome; Déficit immunitaire héréditaire; Candidose cutanéomuqueuse chronique; Primary immune deficiency; Whole exome sequencing; Fungal infection; 616.042
• Online Access: http://www.theses.fr/2017USPCB101/document

Pas de résumé === Chronic mucocutaneous candidiasis (CMC) is seen in human patients with a variety of conditions and refers to recurrent or persistent infection of the skin, nails and/or mucosae by commensal Candida species. Its pathogenesis had long remained elusive, until human genetic studies of rare patients with inherited forms of idiopathic CMC (whether isolated or syndromic), incriminated impaired interleukin (IL)-17A/F immunity. The first genetic etiologies of idiopathic isolated CMC, autosomal dominant (AD) IL-17F and autosomal recessive (AR) IL-17 receptor A (IL-17RA) deficiencies, were reported in 2011 in a multiplex and in a sporadic case, respectively. Using Whole Exome Sequencing (WES), we identified 26 novel patients bearing 15 different homozygous mutations in the IL17RA gene. The mutations identified are either nonsense; missense; frameshift deletions; frameshift insertions; or non-coding essential splice site mutations. Interestingly, 2 alleles encode for surface expressed receptors, whereas all the other tested alleles are not detected at the surface of the patient’s cells (fibroblasts or leucocytes). IL-17RA deficiency is a fully penetrant AR disease, with early onset symptoms, usually within the first year of life. CMC is always present. In addition, 17 patients present with staphylococcal skin infections, and some patients with pyogenic infections of the respiratory tract, including pneumonia. Interestingly, tuberculosis occurred in two unrelated BCG-vaccinated patients. The response to IL-17A and IL-17F homo- and heterodimers is abrogated in fibroblasts, as well as the response to IL-17E/IL-25 in T cells. Human IL-17RA is thus essential for mucocutaneous immunity against Candida and Staphylococcus, but otherwise largely redundant. AR IL-17RA deficiency should be considered in children or adults with CMC, cutaneous staphylococcal disease, or both. In a separate project, I investigated a female child patient born to consanguineous parents who suffered from CMC, recurrent viral infections, disseminated BCG disease and biliary cryptosporidiosis, suggestive of combined immunodeficiency, and who is homozygous for a mutation in REL, encoding the NF-kB protein c-REL. Sanger sequencing confirmed that the patient is homozygous and that both parents are heterozygous for the mutation, consistent with an AR inheritance. The candidate mutation is a nucleotide substitution localized in an acceptor splice site; is not reported in available public databases; and is predicted to be damaging in silico. The mutation disrupts mRNA splicing and is loss-of expression. The patient shows normal counts of lymphoid subsets, with the exception of diminished frequencies of memory CD4+ T, Th2, Th1*, and memory B cells. The patient’s T cells fail to proliferate in response to recall antigens. Naïve CD4+ T cells produce little IL-2 and respond poorly to polyclonal stimulation, a phenotype reverted by exogenous IL-2. Memory CD4+ T cells also produce little amounts of IL-2, and strongly diminished amounts of key effector cytokines (IFN-γ, IL-4, IL-17A and IL-21). The patient exhibited with no detectable specific antibody response following vaccination. Survival and therefore proliferation of naïve B cells are compromised leading to poor generation of plasma cell, and immunoglobulins secretion. The patient shows normal counts of myeloid cells, and frequencies of dendritic cell subsets. IL-12 production is abolished in whole blood in response to BCG+IFN-γ and B-EBV cells in response to mitogens. Although further investigation is needed to fully characterize the patient’s phenotype, these results strongly suggest that the patient suffers from AR complete c-REL deficiency.