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The exposure of autoantigens by microparticles underlies the formation of potent inflammatory components: the microparticle-associated immune complexes.

Cloutier N, Tan S, Boudreau LH, Cramb C, Subbaiah R, Lahey L, Albert A, Shnayder R, Gobezie R, Nigrovic PA, Farndale RW, Robinson WH, Brisson A, Lee DM, Boilard E - EMBO Mol Med (2012)

Bottom Line: Rather, mpICs display autoantigens vimentin and fibrinogen, and recognition of these targets by anti-citrullinated peptide antibodies contributes to the production of mpICs.Functionally, platelet mpICs are highly pro-inflammatory, eliciting leukotriene production by neutrophils.Taken together, our data suggest a unique role for platelet MPs as autoantigen-expressing elements capable of perpetuating formation of inflammatory ICs.

View Article: PubMed Central - PubMed

Affiliation: Faculté de Médecine de l'Université Laval, Centre de Recherche en Rhumatologie et Immunologie, Centre de Recherche du Centre Hospitalier Universitaire de Québec, Québec, Québec, Canada.

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MPs, including platelet MPs, in RA SF form mpICsFSC-PMT and SSC dot plots of representative RA SF labelled with a combination of FITC-conjugated Annexin-V and Cy5-conjugated anti-IgG to demonstrate the presence of IgG on surface of Annexin-V+ MPs. The four-quadrant gates were positioned according to the isotypic controls. The MPs (Annexin-V+-IgG−) in blue show dimensions ranging mostly from 100 to 300 nm (lower inset) while the mpICs (Annexin-V+-IgG+) in green have dimensions from 700 to 3000 nm (upper inset). The relative diameters are presented according to size-defined microsphere calibrations.Quantifications of the mpICs contained in RA and PA SF (n = 23 RA and n = 18 PA ***p < 0.0001).Quantifications of the mpICs relative to the total amounts of detectable ICs in RA SF (n = 23). The statistical analyses are presented under each graph. Data are mean ± SEM.FSC-PMT and SSC dot plots of representative PA SF labelled with a combination of FITC-conjugated Annexin-V and Cy5-conjugated anti-IgG.FSC-PMT and SSC dot plots of RA SF patients labelled with PE-conjugated anti-CD41 and Cy5-conjugated anti-IgG. The four-quadrant gates were positioned according to the isotypic controls. The presence of CD41+ mpICs is evidenced by the dual expression of CD41 and IgG by MPs (green region). RA SF from two patients are presented to illustrate the heterogeneity that exists among the patients.Quantifications of the CD41+ mpICs in RA and PA SF (n = 25 RA and n = 18 PA ***p = 0.0006). The statistical analyses are presented under each graph. Data are mean ± SEM.Triton sensitivity of the CD41+ mpICs contained in RA SF presented as % of untreated (control) CD41+ mpICs.
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fig03: MPs, including platelet MPs, in RA SF form mpICsFSC-PMT and SSC dot plots of representative RA SF labelled with a combination of FITC-conjugated Annexin-V and Cy5-conjugated anti-IgG to demonstrate the presence of IgG on surface of Annexin-V+ MPs. The four-quadrant gates were positioned according to the isotypic controls. The MPs (Annexin-V+-IgG−) in blue show dimensions ranging mostly from 100 to 300 nm (lower inset) while the mpICs (Annexin-V+-IgG+) in green have dimensions from 700 to 3000 nm (upper inset). The relative diameters are presented according to size-defined microsphere calibrations.Quantifications of the mpICs contained in RA and PA SF (n = 23 RA and n = 18 PA ***p < 0.0001).Quantifications of the mpICs relative to the total amounts of detectable ICs in RA SF (n = 23). The statistical analyses are presented under each graph. Data are mean ± SEM.FSC-PMT and SSC dot plots of representative PA SF labelled with a combination of FITC-conjugated Annexin-V and Cy5-conjugated anti-IgG.FSC-PMT and SSC dot plots of RA SF patients labelled with PE-conjugated anti-CD41 and Cy5-conjugated anti-IgG. The four-quadrant gates were positioned according to the isotypic controls. The presence of CD41+ mpICs is evidenced by the dual expression of CD41 and IgG by MPs (green region). RA SF from two patients are presented to illustrate the heterogeneity that exists among the patients.Quantifications of the CD41+ mpICs in RA and PA SF (n = 25 RA and n = 18 PA ***p = 0.0006). The statistical analyses are presented under each graph. Data are mean ± SEM.Triton sensitivity of the CD41+ mpICs contained in RA SF presented as % of untreated (control) CD41+ mpICs.

Mentions: Having visualized mpICs, we employed hs-FCM to analyse them (Fig 3A). We measure 39,400 ± 9400 mpICs/µl in RA SF (Fig 3B). The quantification of the ICs in these same fluids (Supporting Information Fig S1) reveals that the majority (62 ± 7%) of the detectable ICs are in fact mpICs (Fig 3C). These observations provide an explanation for the presence of two subpopulations of MPs evident in RA SF. Specifically larger particles (from ∼700 to 3000 nm; upper inset in Fig 3A) contain mpICs. Conversely, the MPs not associated with IgG are characterized by smaller dimensions (100–300 nm; lower inset in Fig 3A) and represent the majority of the total Annexin-V+ MPs (93% ± 1.4; Supporting Information Fig S2). Interestingly, although MPs and IgG are both present in PA SF, only 2000 ± 900 mpICs/µl could be detected (Fig 3B and D).


The exposure of autoantigens by microparticles underlies the formation of potent inflammatory components: the microparticle-associated immune complexes.

Cloutier N, Tan S, Boudreau LH, Cramb C, Subbaiah R, Lahey L, Albert A, Shnayder R, Gobezie R, Nigrovic PA, Farndale RW, Robinson WH, Brisson A, Lee DM, Boilard E - EMBO Mol Med (2012)

MPs, including platelet MPs, in RA SF form mpICsFSC-PMT and SSC dot plots of representative RA SF labelled with a combination of FITC-conjugated Annexin-V and Cy5-conjugated anti-IgG to demonstrate the presence of IgG on surface of Annexin-V+ MPs. The four-quadrant gates were positioned according to the isotypic controls. The MPs (Annexin-V+-IgG−) in blue show dimensions ranging mostly from 100 to 300 nm (lower inset) while the mpICs (Annexin-V+-IgG+) in green have dimensions from 700 to 3000 nm (upper inset). The relative diameters are presented according to size-defined microsphere calibrations.Quantifications of the mpICs contained in RA and PA SF (n = 23 RA and n = 18 PA ***p < 0.0001).Quantifications of the mpICs relative to the total amounts of detectable ICs in RA SF (n = 23). The statistical analyses are presented under each graph. Data are mean ± SEM.FSC-PMT and SSC dot plots of representative PA SF labelled with a combination of FITC-conjugated Annexin-V and Cy5-conjugated anti-IgG.FSC-PMT and SSC dot plots of RA SF patients labelled with PE-conjugated anti-CD41 and Cy5-conjugated anti-IgG. The four-quadrant gates were positioned according to the isotypic controls. The presence of CD41+ mpICs is evidenced by the dual expression of CD41 and IgG by MPs (green region). RA SF from two patients are presented to illustrate the heterogeneity that exists among the patients.Quantifications of the CD41+ mpICs in RA and PA SF (n = 25 RA and n = 18 PA ***p = 0.0006). The statistical analyses are presented under each graph. Data are mean ± SEM.Triton sensitivity of the CD41+ mpICs contained in RA SF presented as % of untreated (control) CD41+ mpICs.
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fig03: MPs, including platelet MPs, in RA SF form mpICsFSC-PMT and SSC dot plots of representative RA SF labelled with a combination of FITC-conjugated Annexin-V and Cy5-conjugated anti-IgG to demonstrate the presence of IgG on surface of Annexin-V+ MPs. The four-quadrant gates were positioned according to the isotypic controls. The MPs (Annexin-V+-IgG−) in blue show dimensions ranging mostly from 100 to 300 nm (lower inset) while the mpICs (Annexin-V+-IgG+) in green have dimensions from 700 to 3000 nm (upper inset). The relative diameters are presented according to size-defined microsphere calibrations.Quantifications of the mpICs contained in RA and PA SF (n = 23 RA and n = 18 PA ***p < 0.0001).Quantifications of the mpICs relative to the total amounts of detectable ICs in RA SF (n = 23). The statistical analyses are presented under each graph. Data are mean ± SEM.FSC-PMT and SSC dot plots of representative PA SF labelled with a combination of FITC-conjugated Annexin-V and Cy5-conjugated anti-IgG.FSC-PMT and SSC dot plots of RA SF patients labelled with PE-conjugated anti-CD41 and Cy5-conjugated anti-IgG. The four-quadrant gates were positioned according to the isotypic controls. The presence of CD41+ mpICs is evidenced by the dual expression of CD41 and IgG by MPs (green region). RA SF from two patients are presented to illustrate the heterogeneity that exists among the patients.Quantifications of the CD41+ mpICs in RA and PA SF (n = 25 RA and n = 18 PA ***p = 0.0006). The statistical analyses are presented under each graph. Data are mean ± SEM.Triton sensitivity of the CD41+ mpICs contained in RA SF presented as % of untreated (control) CD41+ mpICs.
Mentions: Having visualized mpICs, we employed hs-FCM to analyse them (Fig 3A). We measure 39,400 ± 9400 mpICs/µl in RA SF (Fig 3B). The quantification of the ICs in these same fluids (Supporting Information Fig S1) reveals that the majority (62 ± 7%) of the detectable ICs are in fact mpICs (Fig 3C). These observations provide an explanation for the presence of two subpopulations of MPs evident in RA SF. Specifically larger particles (from ∼700 to 3000 nm; upper inset in Fig 3A) contain mpICs. Conversely, the MPs not associated with IgG are characterized by smaller dimensions (100–300 nm; lower inset in Fig 3A) and represent the majority of the total Annexin-V+ MPs (93% ± 1.4; Supporting Information Fig S2). Interestingly, although MPs and IgG are both present in PA SF, only 2000 ± 900 mpICs/µl could be detected (Fig 3B and D).

Bottom Line: Rather, mpICs display autoantigens vimentin and fibrinogen, and recognition of these targets by anti-citrullinated peptide antibodies contributes to the production of mpICs.Functionally, platelet mpICs are highly pro-inflammatory, eliciting leukotriene production by neutrophils.Taken together, our data suggest a unique role for platelet MPs as autoantigen-expressing elements capable of perpetuating formation of inflammatory ICs.

View Article: PubMed Central - PubMed

Affiliation: Faculté de Médecine de l'Université Laval, Centre de Recherche en Rhumatologie et Immunologie, Centre de Recherche du Centre Hospitalier Universitaire de Québec, Québec, Québec, Canada.

Show MeSH
Related in: MedlinePlus