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Leucine-Rich Repeat Kinase 2 (Lrrk2) Deficiency Diminishes the Development of Experimental Autoimmune Uveitis (EAU) and the Adaptive Immune Response.

Wandu WS, Tan C, Ogbeifun O, Vistica BP, Shi G, Hinshaw SJ, Xie C, Chen X, Klinman DM, Cai H, Gery I - PLoS ONE (2015)

Bottom Line: In both these diseases inflammatory processes participate in the pathogenic process.Peritoneal macrophages were examined for their production of cytokines/chemokines in culture following stimulation with LPS or the oligodeoxynucleotide CpG.The expression levels of FoxP3 by Lrrk2 (-/-) spleen cells, however, were similar to those seen in WT controls.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunology, National Eye Institute, National Institutes of Health, Bethesda, MD, 20892, United States of America.

ABSTRACT

Background: Mutations in LRRK2 are related to certain forms of Parkinson's disease and, possibly, to the pathogenesis of Crohn's disease. In both these diseases inflammatory processes participate in the pathogenic process. LRRK2 is expressed in lymphoid cells and, interestingly, Lrrk2 (-/-) mice were reported to develop more severe experimental colitis than their wild type (WT) controls. Here, we examined the possible involvement of LRRK2 in the pathogenesis of experimental autoimmune uveitis (EAU), an animal model for human uveitis, by testing Lrrk2 (-/-) mice for their capacity to develop this experimental eye disease and related immune responses.

Methods: Lrrk2 (-/-) mice and their WT controls (C57Bl/6) were immunized with interphotoreceptor retinoid-binding protein (IRBP) and compared for their development of EAU, delayed type hypersensitivity (DTH) by skin tests, production of cytokines in culture, and expression of interferon (IFN)-γ, interleukin (IL)-17 and FoxP3 by spleen cells, using flow cytometry. Peritoneal macrophages were examined for their production of cytokines/chemokines in culture following stimulation with LPS or the oligodeoxynucleotide CpG. The Lrrk2 (-/-) and WT mice were also compared for their response to bovine serum albumin (BSA).

Results: The Lrrk2 (-/-) mice developed lower levels of EAU, DTH responses and cytokine production by lymphocytes than did their WT controls. Intracellular expression of IFN-γ and IL-17, by spleen cells, and secretion of cytokines/chemokines by activated peritoneal macrophages of Lrrk2 (-/-) mice trended toward diminished levels, although variabilities were noted. The expression levels of FoxP3 by Lrrk2 (-/-) spleen cells, however, were similar to those seen in WT controls. Consistent with their low response to IRBP, Lrrk2 (-/-) mice responded to BSA less vigorously than their WT controls.

Conclusions: Lrrk2 deficiency in mice diminished the development of EAU and the related adaptive immune responses to IRBP as compared to the WT controls.

No MeSH data available.


Related in: MedlinePlus

Minute differences seen in the profiles of cytokines/chemokines released by peritoneal macrophages of the Lrrk2 (-/-) (“KO”) and their WT controls, cultured with LPS (0.5 μg/ml) or CpG (40 μg/ml) for 48 hrs.The levels of the released products were measured by a bead-based multi-plex screening assay. The assay measured secretion of 14 analytes, as detailed in the Materials and Methods section, but molecules with undetectable levels were not included in the figure. The figures show combined data collected in two individual experiments and combined for presentation.
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pone.0128906.g005: Minute differences seen in the profiles of cytokines/chemokines released by peritoneal macrophages of the Lrrk2 (-/-) (“KO”) and their WT controls, cultured with LPS (0.5 μg/ml) or CpG (40 μg/ml) for 48 hrs.The levels of the released products were measured by a bead-based multi-plex screening assay. The assay measured secretion of 14 analytes, as detailed in the Materials and Methods section, but molecules with undetectable levels were not included in the figure. The figures show combined data collected in two individual experiments and combined for presentation.

Mentions: In addition to their participation in phagocytosis, macrophages play major roles in the immune system as producers of cytokines and chemokines. To compare between macrophages of Lrrk2 deficient mice and WT controls, we collected their induced peritoneal macrophages and measured the levels of certain cytokines and chemokines released in cultures stimulated with two TLR ligands, LPS and CpG [22]. The data collected in two separate experiments are combined in Fig 5 and show only small differences in levels of the majority of tested molecules produced by macrophages of the two mouse groups when stimulated by LPS or CpG. These observations are in line with findings of a study by Dzamko, et al [23]. It is of interest, however, that the mean levels of TNF-α were higher in supernatants of WT than of Lrrk2 (-/-) cells when cultured with either LPS or CpG.


Leucine-Rich Repeat Kinase 2 (Lrrk2) Deficiency Diminishes the Development of Experimental Autoimmune Uveitis (EAU) and the Adaptive Immune Response.

Wandu WS, Tan C, Ogbeifun O, Vistica BP, Shi G, Hinshaw SJ, Xie C, Chen X, Klinman DM, Cai H, Gery I - PLoS ONE (2015)

Minute differences seen in the profiles of cytokines/chemokines released by peritoneal macrophages of the Lrrk2 (-/-) (“KO”) and their WT controls, cultured with LPS (0.5 μg/ml) or CpG (40 μg/ml) for 48 hrs.The levels of the released products were measured by a bead-based multi-plex screening assay. The assay measured secretion of 14 analytes, as detailed in the Materials and Methods section, but molecules with undetectable levels were not included in the figure. The figures show combined data collected in two individual experiments and combined for presentation.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4465928&req=5

pone.0128906.g005: Minute differences seen in the profiles of cytokines/chemokines released by peritoneal macrophages of the Lrrk2 (-/-) (“KO”) and their WT controls, cultured with LPS (0.5 μg/ml) or CpG (40 μg/ml) for 48 hrs.The levels of the released products were measured by a bead-based multi-plex screening assay. The assay measured secretion of 14 analytes, as detailed in the Materials and Methods section, but molecules with undetectable levels were not included in the figure. The figures show combined data collected in two individual experiments and combined for presentation.
Mentions: In addition to their participation in phagocytosis, macrophages play major roles in the immune system as producers of cytokines and chemokines. To compare between macrophages of Lrrk2 deficient mice and WT controls, we collected their induced peritoneal macrophages and measured the levels of certain cytokines and chemokines released in cultures stimulated with two TLR ligands, LPS and CpG [22]. The data collected in two separate experiments are combined in Fig 5 and show only small differences in levels of the majority of tested molecules produced by macrophages of the two mouse groups when stimulated by LPS or CpG. These observations are in line with findings of a study by Dzamko, et al [23]. It is of interest, however, that the mean levels of TNF-α were higher in supernatants of WT than of Lrrk2 (-/-) cells when cultured with either LPS or CpG.

Bottom Line: In both these diseases inflammatory processes participate in the pathogenic process.Peritoneal macrophages were examined for their production of cytokines/chemokines in culture following stimulation with LPS or the oligodeoxynucleotide CpG.The expression levels of FoxP3 by Lrrk2 (-/-) spleen cells, however, were similar to those seen in WT controls.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunology, National Eye Institute, National Institutes of Health, Bethesda, MD, 20892, United States of America.

ABSTRACT

Background: Mutations in LRRK2 are related to certain forms of Parkinson's disease and, possibly, to the pathogenesis of Crohn's disease. In both these diseases inflammatory processes participate in the pathogenic process. LRRK2 is expressed in lymphoid cells and, interestingly, Lrrk2 (-/-) mice were reported to develop more severe experimental colitis than their wild type (WT) controls. Here, we examined the possible involvement of LRRK2 in the pathogenesis of experimental autoimmune uveitis (EAU), an animal model for human uveitis, by testing Lrrk2 (-/-) mice for their capacity to develop this experimental eye disease and related immune responses.

Methods: Lrrk2 (-/-) mice and their WT controls (C57Bl/6) were immunized with interphotoreceptor retinoid-binding protein (IRBP) and compared for their development of EAU, delayed type hypersensitivity (DTH) by skin tests, production of cytokines in culture, and expression of interferon (IFN)-γ, interleukin (IL)-17 and FoxP3 by spleen cells, using flow cytometry. Peritoneal macrophages were examined for their production of cytokines/chemokines in culture following stimulation with LPS or the oligodeoxynucleotide CpG. The Lrrk2 (-/-) and WT mice were also compared for their response to bovine serum albumin (BSA).

Results: The Lrrk2 (-/-) mice developed lower levels of EAU, DTH responses and cytokine production by lymphocytes than did their WT controls. Intracellular expression of IFN-γ and IL-17, by spleen cells, and secretion of cytokines/chemokines by activated peritoneal macrophages of Lrrk2 (-/-) mice trended toward diminished levels, although variabilities were noted. The expression levels of FoxP3 by Lrrk2 (-/-) spleen cells, however, were similar to those seen in WT controls. Consistent with their low response to IRBP, Lrrk2 (-/-) mice responded to BSA less vigorously than their WT controls.

Conclusions: Lrrk2 deficiency in mice diminished the development of EAU and the related adaptive immune responses to IRBP as compared to the WT controls.

No MeSH data available.


Related in: MedlinePlus