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Reduced type I interferon production by dendritic cells and weakened antiviral immunity in patients with Wiskott-Aldrich syndrome protein deficiency.

Lang PA, Shaabani N, Borkens S, Honke N, Scheu S, Booth S, Brenner D, Meryk A, Barthuber C, Recher M, Mak TW, Ohashi PS, Häussinger D, Griffiths GM, Thrasher AJ, Bouma G, Lang KS - J. Allergy Clin. Immunol. (2012)

Bottom Line: Despite a significant proportion of patients with WAS having recurrent viral infections, surprisingly little is known about the effects of WASP deficiency on antiviral immunity.IFN-I production by WAS KO DCs was reduced both in vivo and in vitro.These findings might help us to understand the immunodeficiency of WAS.

View Article: PubMed Central - PubMed

Affiliation: Campbell Family Institute for Breast Cancer Research, Ontario Cancer Institute, Toronto, Ontario, Canada.

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Total number of leukocytes in lymphoid tissue. The number of splenic CD11c+mPDCA1+ pDCs (A) and the total number of leukocytes was determined in spleens (B) and lymph nodes (C). DC subsets were quantified in spleens (D) and lymph nodes (E). IFN-α expression was determined by means of intracellular flow cytometry after stimulation of ex vivo–isolated DC subsets (F). Symbols in Fig E2, A to C, represent individual mice, and the line indicates the mean. Data in Fig E2, D to F, are expressed as means ± SEMs of n = 4. Fig E2, F, LPS and CpG, n = 7; Poly(I:C), n = 4.
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dfig2: Total number of leukocytes in lymphoid tissue. The number of splenic CD11c+mPDCA1+ pDCs (A) and the total number of leukocytes was determined in spleens (B) and lymph nodes (C). DC subsets were quantified in spleens (D) and lymph nodes (E). IFN-α expression was determined by means of intracellular flow cytometry after stimulation of ex vivo–isolated DC subsets (F). Symbols in Fig E2, A to C, represent individual mice, and the line indicates the mean. Data in Fig E2, D to F, are expressed as means ± SEMs of n = 4. Fig E2, F, LPS and CpG, n = 7; Poly(I:C), n = 4.

Mentions: To investigate which cells were responsible for the defective production of IFN-Is, we made use of the IFN-β reporter–knock-in mouse, in which yellow fluorescent protein (YFP) expression is bicistronically linked to expression of IFN-β of the endogenous ifnb locus, so that IFN-β–producing cells can easily be identified by using YFP expression.46 These IFNβmob/mob mice were crossed with WAS KO mice and challenged with Poly(I:C). As expected, we found that in the absence of WASP, IFN-β/YFP expression was reduced in splenocytes and that this was restricted to CD11c+ cells (Fig 6, A and B). To verify that this was not caused by an overall reduction in the number of DCs in WAS KO mice, we analyzed the proportion of conventional migratory (cDCs; CD11c+CD11b+CD8α−B220−), conventional CD8α+ (CD11c+CD11b−CD8α+B220−), and plasmacytoid (pDCs; CD11c+CD11b−B220+ or CD11c+mPDCA1+) DC subsets. As expected from previous reports,14,47 we did not observe significant differences between distinct subsets in spleens or lymph nodes in C57BL/6 or WAS KO mice (Fig 6, C and D, and see Fig E2, A, in this article's Online Repository at www.jacionline.org). Total splenocyte and lymph node cell numbers in WAS KO and C57BL/6 mice were comparable, as were absolute cell counts of DC subsets (see Fig E2, B-E). Finally, we tested whether the impaired production of IFN-I observed in vivo reflected an intrinsic deficiency of DCs in the absence of WASP. Both pDCs and cDCs showed a reduced IFN-α response when stimulated with Poly(I:C), CpG, and LPS in vitro (Fig 6, E). Similarly, when we used ex vivo–isolated splenic CD11c+ cells, a similar deficiency in IFN-α production in response to Poly(I:C), LPS, and CpG was observed (see Fig E2, F). These findings show that WAS KO DCs are intrinsically compromised in their ability to secrete IFN-I.


Reduced type I interferon production by dendritic cells and weakened antiviral immunity in patients with Wiskott-Aldrich syndrome protein deficiency.

Lang PA, Shaabani N, Borkens S, Honke N, Scheu S, Booth S, Brenner D, Meryk A, Barthuber C, Recher M, Mak TW, Ohashi PS, Häussinger D, Griffiths GM, Thrasher AJ, Bouma G, Lang KS - J. Allergy Clin. Immunol. (2012)

Total number of leukocytes in lymphoid tissue. The number of splenic CD11c+mPDCA1+ pDCs (A) and the total number of leukocytes was determined in spleens (B) and lymph nodes (C). DC subsets were quantified in spleens (D) and lymph nodes (E). IFN-α expression was determined by means of intracellular flow cytometry after stimulation of ex vivo–isolated DC subsets (F). Symbols in Fig E2, A to C, represent individual mice, and the line indicates the mean. Data in Fig E2, D to F, are expressed as means ± SEMs of n = 4. Fig E2, F, LPS and CpG, n = 7; Poly(I:C), n = 4.
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dfig2: Total number of leukocytes in lymphoid tissue. The number of splenic CD11c+mPDCA1+ pDCs (A) and the total number of leukocytes was determined in spleens (B) and lymph nodes (C). DC subsets were quantified in spleens (D) and lymph nodes (E). IFN-α expression was determined by means of intracellular flow cytometry after stimulation of ex vivo–isolated DC subsets (F). Symbols in Fig E2, A to C, represent individual mice, and the line indicates the mean. Data in Fig E2, D to F, are expressed as means ± SEMs of n = 4. Fig E2, F, LPS and CpG, n = 7; Poly(I:C), n = 4.
Mentions: To investigate which cells were responsible for the defective production of IFN-Is, we made use of the IFN-β reporter–knock-in mouse, in which yellow fluorescent protein (YFP) expression is bicistronically linked to expression of IFN-β of the endogenous ifnb locus, so that IFN-β–producing cells can easily be identified by using YFP expression.46 These IFNβmob/mob mice were crossed with WAS KO mice and challenged with Poly(I:C). As expected, we found that in the absence of WASP, IFN-β/YFP expression was reduced in splenocytes and that this was restricted to CD11c+ cells (Fig 6, A and B). To verify that this was not caused by an overall reduction in the number of DCs in WAS KO mice, we analyzed the proportion of conventional migratory (cDCs; CD11c+CD11b+CD8α−B220−), conventional CD8α+ (CD11c+CD11b−CD8α+B220−), and plasmacytoid (pDCs; CD11c+CD11b−B220+ or CD11c+mPDCA1+) DC subsets. As expected from previous reports,14,47 we did not observe significant differences between distinct subsets in spleens or lymph nodes in C57BL/6 or WAS KO mice (Fig 6, C and D, and see Fig E2, A, in this article's Online Repository at www.jacionline.org). Total splenocyte and lymph node cell numbers in WAS KO and C57BL/6 mice were comparable, as were absolute cell counts of DC subsets (see Fig E2, B-E). Finally, we tested whether the impaired production of IFN-I observed in vivo reflected an intrinsic deficiency of DCs in the absence of WASP. Both pDCs and cDCs showed a reduced IFN-α response when stimulated with Poly(I:C), CpG, and LPS in vitro (Fig 6, E). Similarly, when we used ex vivo–isolated splenic CD11c+ cells, a similar deficiency in IFN-α production in response to Poly(I:C), LPS, and CpG was observed (see Fig E2, F). These findings show that WAS KO DCs are intrinsically compromised in their ability to secrete IFN-I.

Bottom Line: Despite a significant proportion of patients with WAS having recurrent viral infections, surprisingly little is known about the effects of WASP deficiency on antiviral immunity.IFN-I production by WAS KO DCs was reduced both in vivo and in vitro.These findings might help us to understand the immunodeficiency of WAS.

View Article: PubMed Central - PubMed

Affiliation: Campbell Family Institute for Breast Cancer Research, Ontario Cancer Institute, Toronto, Ontario, Canada.

Show MeSH
Related in: MedlinePlus