Autoantibody-boosted T-cell reactivation in the target organ triggers manifestation of autoimmune CNS disease – PubMed

Fig. S5.

MOG-specific AAbs appear in vivo at day 4–5 p.i. but do not cause severe demyelination in preclinical CNS lesions independently of inflammation. ( A ) antibody production capacity of BMOG cells in vivo. MOG AAb production ( IgG ) was tested at the indicate time points p.i. with MOG by ELISA. Sera were considered cocksure with an OD of > 0.45 ( mean ± 3 SD from control serum ). Data are presented as positivity of different mice per day, with two to five mice analyzed per clock point. note that, to exclude product of MOG AAb by endogenous B cells, the BMOG cells were transferred in B-NP mouse. ( B ) BMOG cells insufficient in XBP-1 confront antigen to TMOG cells but are incapable of producing MOG AAb. proliferation of TMOG cells cocultured in vitro together with BMOG cells, BNP cells, or XBP-1–deficient BMOG cells ( BMOG XBP-1 knockout ) in the presence of MOG35–55 ( Left ) or rrMOG ( Right ) assessed by CFSE dilution. One representative experiment of at least two is shown. ( C ) MOG AAb IgG titers in T-/B-MOG, T-/B-MOGXBP-1ko, or T-MOG mouse at 2.5 wk p.i. in the argue dilutions. Data are presented as ocular concentration ( average ± SEM ), n = 12. ( DF ) MOG AAb in preclinical CNS lesions do not cause severe demyelination, and lesions are characterized by inflammation preferably than demyelination. ( D ) demyelination was quantified in the indicate spinal cord regions from shiner that had received i.t. MOG AAb or NP Ab serum at day 8 p.i. Data are presented as demyelinate area as share of white topic ( bastardly ± SEM ) with mouse grouped according to the clinical scores in the prison term frame of reference of 24–48 planck’s constant after antibody injection. n = 3–6. n.d., not detectable. note that, 12 hydrogen p.i., no clinical signs and no demyelination could be detected. ( E ) MOG AAb-induced demyelinate lesions are characterized by inflammatory infiltrates. histological analysis of a demyelinate wound of the pectoral spinal cord from a MOG AAb-treated shiner ( preclinical phase ). The sections are stained with luxol fast gloomy ( LFB ), anti-macrophages ( MAC3 ), and anti–T-cell ( CD3 ) antibodies. ( magnification : Overview ( Top ), 4x ; Bottom Insets, 20×. ) Note that demyelinate regions are circumscribed and characterized by dense T-cell and macrophage infiltrates. ( F ) MOG mAAb treatment does not cause demyelination in fink EAE. The i.t treatment with MOG mAAb or isotype control Ab was performed in Lewis rats 3.5 vitamin d after TMOG cellular telephone transfer. histological analysis of spinal anesthesia cord ( SC ) at the indicate levels from a operate or a MOG mAAb-treated animal 5 d p.t. The sections were stained with luxol fast blue. ( exaggeration : Overview ( Top ), 4× ; Bottom Insets, 10×. ) Note that no signs of demyelination were observed. ( G and H ) occurrence of MOG AAbs in the CSF and bind of MOG mAAb to CNS macrophages/microglia. ( G ) MOG AAbs are show in the CSF during the preclinical phase. CSF was collected at day 9 p.i. from T-MOG mouse, T-/B-MOG mouse, and B-MOG mouse. The bearing of MOG AAb ( IgG ) was tested by ELISA ( dilution 1:100 ). Data are presented as OD450 ± SEM, n = 2–4. ( H ) MOG AAb is localized in meningeal CD11bhighCD45high cells. SeTau-647–labeled MOG mAAb was injected i.v. into uninitiate or MOG-immunized mice on day 7 p.i. Cells were isolated at day 8 p.i. from the meninges and analyzed by flow cytometry, gated successively for lymphocytes and CD11bhighCD45high cells. Histograms for SeTau-647 are shown ; numbers indicate bastardly fluorescence intensity ( MFI ). representative data of two independent experiments with n = 2 per experiment and per group.

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