Defective Leukocyte Adhesion and Chemotaxis Contributes to Combined Immunodeficiency in Humans with Autosomal Recessive MST1 Deficiency - PubMed (original) (raw)

Case Reports

doi: 10.1007/s10875-016-0232-2. Epub 2016 Jan 22.

Joseph D P Willet 1, Helen R Griffin 2, Neil V Morgan 3, Graeme O'Boyle 1, Peter D Arkwright 4, Stephen M Hughes 4, Mario Abinun 1 5, Louise J Tee 3, Dawn Barge 6, Karin R Engelhardt 1, Michael Jackson 5, Andrew J Cant 1 5, Eamonn R Maher 3 7, Mauro Santibanez Koref 2, Louise N Reynard 1, Simi Ali 1, Sophie Hambleton 8 9

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Case Reports

Defective Leukocyte Adhesion and Chemotaxis Contributes to Combined Immunodeficiency in Humans with Autosomal Recessive MST1 Deficiency

Tarana Singh Dang et al. J Clin Immunol. 2016 Feb.

Erratum in

Abstract

Purpose: To investigate the clinical and functional aspects of MST1 (STK4) deficiency in a profoundly CD4-lymphopenic kindred with a novel homozygous nonsense mutation in STK4. Although recent studies have described the cellular effects of murine Mst1 deficiency, the phenotype of MST1-deficient human lymphocytes has yet to be fully explored. Patient lymphocytes were therefore investigated in the context of current knowledge of murine Mst1 deficiency.

Methods: Genetic etiology was identified by whole exome sequencing of genomic DNA from two siblings, combined with linkage analysis in the wider family. MST1 protein expression was assessed by immunoblotting. The ability of patient lymphocytes to adhere to ICAM-1 under flow conditions was measured, and transwell assays were used to assess chemotaxis. Chemokine receptor expression was examined by flow cytometry and receptor signalling by immunoblotting.

Results: A homozygous nonsense mutation in STK4 (c.442C > T, p.Arg148Stop) was found in the patients, leading to a lack of MST1 protein expression. Patient leukocytes exhibited deficient chemotaxis after stimulation with CXCL11, despite preserved expression of CXCR3. Patient lymphocytes were also unable to bind effectively to immobilised ICAM-1 under flow conditions, in keeping with a failure to develop high affinity binding.

Conclusion: The observed abnormalities of adhesion and migration imply a profound trafficking defect among human MST1-deficient lymphocytes. By analogy with murine Mst1 deficiency and other defects of leucocyte trafficking, this is likely to contribute to immunodeficiency by impairing key aspects of T-cell development and function such as positive selection in the thymus, thymic egress and immune synapse formation in the periphery.

Keywords: CD4 lymphopenia; Combined immunodeficiency; MST1; STK4; chemotaxis; lymphocyte adhesion.

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Figures

Fig. 1

Fig. 1

a Pedigree of affected family, demonstrating consanguinity (double horizontal lines). Males and females are indicated by squares and circles respectively, with open symbols indicating healthy individuals and shaded symbols indicating affected individuals. Triangles indicate unaffected individuals with censored gender, and diagonal lines indicate deceased individuals. b Sanger sequencing of STK4 gene confirmed WES findings – a homozygous C to T substitution at position 442 in patients that was heterozygous in both parents. c Western blotting of P3-derived EBV-transformed B cell lysate and P1 and P3 fibroblast lysates, demonstrating MST1 deficiency. Control lysates were derived from random healthy individuals. Representative of a minimum of two separate experiments

Fig. 2

Fig. 2

a Patient CD4+ T lymphocytes exhibit expression of the CXCL11 receptor CXCR3 (red histogram = unstained control, green histogram = anti-CXCR3-PE antibody) (n = 1). b EBV-transformed B cells were treated with 12 nM CXCL11 to stimulate CXCR3 before whole cell lysis, Western transfer analysis and probing for phospho-ERK, total ERK, phospho-AKT and total AKT, with β-tubulin as a loading control. Numbers indicate time (mins) of treatment with CXCL11. Representative of three separate experiments. c CXCL11 treated patient EBV-transformed B cells (left panel) show reduced binding to ICAM-1-coated chips under flow conditions when compared to control cells (* indicates P = 0.0313). Patient PBLs also show reduced binding when compared to control cells (right panel). Each shape represents one of six separate experiments, with 5 replicates for each. Significance was determined using Wilcoxon signed rank tests. d CXCL11-treated patient PBLs show unchanged capacity for migration across transwell filters when compared to control cells (n = 1)

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