Summary of clinical neurological presentations.

<p>Summary of clinical neurological presentations.</p

Functional study of the IFN-γ/IL-12 axis and oxidative burst in P1.

<p>A). Whole-blood IL-12/IFN-γ pathway screening. P1 and P2 had normal responses to IFN-γ and IL-12. B). EMSA detecting GAF DNA-binding activity after IFN-γ stimulation in the EBV-B cells from two healthy controls, P1 and individuals with complete recessive IFN-γR2 deficiency, used as a negative control. Cells were stimulated for 15 minutes with the indicated dose of IFN-γ. C). Oxidative burst. The production of superoxide in the EBV-B cells from healthy controls, P1 and patients with chronic granulomatous disease, used as negative controls, was determined following stimulation with PMA. These experiments were carried out at least twice.</p

Immunoprecipitation and immunofluorescence of the AP-4 complex in P1.

<p>A). EBV-B cells and SV40 fibroblasts from P1 and a healthy control were subjected to immunoprecipitation under native conditions with antibodies against AP-4ε or AP-4β, and the immunoprecipitates were then western blotted with antibodies against AP-4ε or AP-4β. Equal amounts of protein were used for immunoprecipitation from patient and control cells. A small amount of AP-4β is coassembled with AP-4ε, and the AP-4ε assembled with AP-4β in P1 appears to be slightly smaller (AP-4ε*) than the AP-4ε in control cells. B). Fibroblasts from P1 and a healthy control were double-labeled for AP-4ε and the AP-4-associated protein tepsin. Note the specific loss of AP-4ε and tepsin from the cells of P1. Bar 20 µm. These experiments were carried out at least three times.</p

Summary of whole-exome sequencing results.

a<p>Number of variants not found in dbSNP or 1000 Genomes or HapMap and <0.001% in our database;</p>b<p>Hom: homozygous mutation;</p>c<p>Het, heterozygous mutation;</p>d<p>UTR-5: the five-prime untranslated region;</p>e<p>UTR-3: the three-prime untranslated region;</p>f<p>lincRNA: long non-coding RNA;</p>g<p>miRNA: microRNA.</p

mRNA and protein levels for the subunits of the AP-4 complex.

<p>A). RT-qPCR to assess mRNA levels for the components of the AP-4 complex in EBV-B cells from P1. B). RT-PCR to assess the splicing of <i>AP4E1</i> mRNA. C). Western blot: whole-cell homogenates from EBV-B cells from P1 and a healthy control were subjected to western blotting for clathrin heavy chain (CHC; loading control), AP-4ε, AP-4β or AP-4 μ. The loss of AP-4ε results in a concomitant decrease in the levels of AP-4β and AP-4 μ (specific bands are indicated by an arrow). These experiments were carried out at least twice.</p

Table indicating the clinical presentations of TB recorded in the fifty patients.

<p>The two patients presenting with IL-12Rβ1 deficiency are in bold. N means number and age is indicated in years.</p

Mendelian mutations in <i>IL12RB1</i> leading to severe tuberculosis in two kindreds.

<p><b>A</b>. Pedigree of the two families (A and B) with IL-12Rβ1 deficiency. Each generation is designated by a roman numeral (I–II), and each individual by an Arabic numeral. The double lines connecting the parents indicate consanguinity. The probands are indicated by an arrow, with black indicating <i>Mycobacterium tuberculosis</i> disease status. Individuals whose genetic status could not be evaluated are indicated by the symbol “E?”. <b>B</b>. Electrophoregram showing the genomic sequences of exons 9 and 5 in patients 1 and 2, respectively, compared with a control sequence. <b>C</b>. Schematic diagram of the coding region of the IL-12Rβ1 chain containing 17 coding exons and encoding a 662-amino acid protein with a leader sequence (exon1, L), extracellular domain (exons 2 to 13, EC), transmembrane domain (exon 14, TM) and an intracellular cytoplasmic domain (exons 15 to 17, IC). Published and unpublished mutations are indicated as follows: missense mutations are shown in purple, nonsense mutations are shown in red and complex mutations are shown in brown. Splicing mutations are shown in blue, large deletions are shown in green, insertions are shown in orange, and duplication is shown in magenta. * The 700+362_1619-944del mutation is the only mutation resulting in at the expression of a protein at the cell surface. Mutations of P1 (K305X) and P2 (R173W) are underlined. <b>D</b>. Chest X ray of patient 1 showing the localization of the disease.</p