Journal: bioRxiv

Article Title: Divergent and dominant anti-inflammatory effects of patient-derived anti-citrullinated protein antibodies (ACPA) in arthritis development

doi: 10.1101/2022.09.30.510377

Figure Lengend Snippet: (A) Schematic representation of the animal model used. (B) Anti-citrullinated peptide and protein reactivity by monoclonal ACPAs used in the study. (C-I) CAIA was induced in mice by i.v. transfer of arthritogenic anti-CII antibody cocktail. Monoclonal ACPAs (C, D and E), C03 F(ab’)2 fragments (G), mutated FcγR-null GRLR-C03 ACPA (H) or polyclonal CCP-pool and respective flow-through (I) were transferred simultaneously at time of disease induction. Therapeutic effect of ACPAs was assessed by transfer of C03 ACPA at the peak of disease, day 8 (F). Boosting and synchronization of disease symptoms was done by i.p. administration of LPS 3 days post disease induction. Statistical analysis calculated by non-parametric repeated-measures Friedman test with Dunn’s multiple comparison test. Disease curves from mC03 and mE02 reference groups are identical in panels C and D due to splitting of the data into two panels for better visualization. Cit: citrullination; KAc: acetylated lysine; Carb: carbamylation; TNC: tenascin C; Fib: fibrinogen; Vim: vimentin; Fil: fillagrin; His4: histone 4; mC03: murine IgG2a C03; hC03: human IgG1 C03; FT: flow-through; IVIG: intravenous IgG preparation.

Article Snippet: ACPA mAbs were screened for binding to citrullinated, native control peptides and control antigens using a custom-designed antigen microarray (ThermoFisher Scientific, Immunodiagnostics) .

Techniques: Animal Model

Journal: Acta neuropathologica

Article Title: Frontal cortex and striatal cellular and molecular pathobiology in individuals with Down syndrome with and without dementia

doi: 10.1007/s00401-019-01965-6

Figure Lengend Snippet: List of antibodies

Article Snippet: To validate custom-designed microarray expression data, additional FC sections from both groups were incubated with a rabbit polyclonal antibody against the human nuclear sirtuin 6 (SIRT 6; 1:5000, Sigma), after performing antigen retrieval using Dako target retrieval solution, pH 9 (DAKO, Carpinteria, CA) for 20 minutes in a steamer [ 96 ]. table ft1 table-wrap mode="anchored" t5 Table 2. caption a7 Antigen Antibody Dilution IH (IF) Company Catalog # APP/Aβ (6E10) IgG1 mouse monoclonal to residues 1–16 of Aβ 1:1000 BioLegend #803002 Aβ (MOAB2) IgG2b mouse monoclonal to Aβ 1:800 Gift from M. J. LaDu Aβ42 IgG rabbit polyclonal to Aβ42 1:1000 Millipore #AB5078 Conformational Tau (Alz50) IgM mouse monoclonal to tau residues 5–15, 312–322 1:1000 (1:50) Gift from P. Davies Conformational Tau (MC1) IgG1 mouse monoclonal to tau residues 5–15, 312–322 1:1000 Gift from P. Davies Phosphorylated Tau (pS422) IgG rabbit polyclonal to tau phospho-Serine 422 1:1000 (1:50) Invitrogen # 03151 Phosphorylated Tau (AT8) IgG1 mouse monoclonal to tau phospo-Serine 202 and phospho-Threonine 205 1:1000 (1:50) Invitrogen # MN1020 Truncated Tau (TauC3) IgG1 mouse monoclonal to truncated tau at 421/422 residues 1:1000 (1:50) ThermoFisher # AHB0061 Truncated Tau (MN423) IgG1 mouse monoclonal to truncated tau at glutamic acid 391 1:5000 Gift from L. Binder Choline Acetyltransferase (ChAT) IgG goat polyclonal to human placental choline acetyltransferase 1:800 (1:50) Millipore #AB144P Tyrosine Hydroxylase (TH) IgG rabbit polyclonal to tyrosine hydroxylase 1:1000 Invitrogen #OPA1–04050 Neurofilament (SMI-34) IgG1 mouse monoclonal to phosphorylated neurofilament H 1:1000 BioLegend #8535503 Sirtuin 6 IgG rabbit polyclonal to amino acids 330–348 of human Sirt6 1:5000 Sigma #S4197 Open in a separate window IH, Immunohistochemistry; IF, Immunofluorescence List of antibodies To visualize the relationship between amyloid plaques and tau-containing neurites, additional cortical and striatal sections were dual immunolabeled using two different colored chromogens.

Techniques:

Journal: Acta neuropathologica

Article Title: Frontal cortex and striatal cellular and molecular pathobiology in individuals with Down syndrome with and without dementia

doi: 10.1007/s00401-019-01965-6

Figure Lengend Snippet: Photomicrographs of dual labeled frontal cortex sections showing dystrophic neurites displaying immunoreactivity for the tau conformational epitope MC1 (brown) intermingled within Aβ42-ir plaques (blue) in a 47 year-old female non-demented (a) and a 46 year-old male demented (d) individual with DS. Note the presence of numerous MC1-ir NTs (small arrows) in the demented compared to non-demented DS case. b, c and e, High-power images of plaques (large arrows in a and d) displaying dystrophic neurites. Images of frontal cortex showing Aβ42-ir plaques (blue) surrounded by dystrophic neurites positive for the phosphorylated neurofilament H marker SMI-34 (dark brown) in a 46 year-old male demented person with DS (f). g and h, High-power images showing the bulbous nature of the dystrophic neurites within Aβ42-ir plaques shown in panel f (arrows). Photomicrographs of the putamen showing Aβ42-ir plaques (dark blue) and neuropil lacking MC1-ir dystrophic neurites (brown) in a 47 year-old female non-demented individual with DS (i). Inset shows high-power image of Aβ42-ir plaque from panel i (arrow). j. Image of a dual labeled caudate section demonstrating the absence of SMI-34-ir (dark brown) dystrophic neurites within Aβ42-ir plaques (blue) in a 46 year-old male demented subject with DS. Inset show an apparent intact SMI-34-ir fibers (arrow) in close proximity to an Aβ42-ir plaque (j). k and l, Images showing Aβ42-ir plaques (dark blue) and ChAT-ir neurons (brown) (k) as well as APP/Aβ-ir plaques (blue) and TH-ir fibers (brown) (l) in the putamen of a 46 year-old male demented subject with DS. Note the lack of ChAT-ir (k) and TH-ir (l) dystrophic neurites within plaques. Insets in k and l images show the absence of morphological alterations in a ChAT positive interneuron adjacent to an Aβ42-ir plaque as well as TH-ir fibers neighboring an APP/Aβ-ir plaque indicated (arrows), respectively. Abbreviations, DSD−, DS without dementia, DSD+, DS with dementia. Scale bars in a, d, f and i-l = 50 μm and c, e, g, h and insets =10 μm.

Article Snippet: To validate custom-designed microarray expression data, additional FC sections from both groups were incubated with a rabbit polyclonal antibody against the human nuclear sirtuin 6 (SIRT 6; 1:5000, Sigma), after performing antigen retrieval using Dako target retrieval solution, pH 9 (DAKO, Carpinteria, CA) for 20 minutes in a steamer [ 96 ]. table ft1 table-wrap mode="anchored" t5 Table 2. caption a7 Antigen Antibody Dilution IH (IF) Company Catalog # APP/Aβ (6E10) IgG1 mouse monoclonal to residues 1–16 of Aβ 1:1000 BioLegend #803002 Aβ (MOAB2) IgG2b mouse monoclonal to Aβ 1:800 Gift from M. J. LaDu Aβ42 IgG rabbit polyclonal to Aβ42 1:1000 Millipore #AB5078 Conformational Tau (Alz50) IgM mouse monoclonal to tau residues 5–15, 312–322 1:1000 (1:50) Gift from P. Davies Conformational Tau (MC1) IgG1 mouse monoclonal to tau residues 5–15, 312–322 1:1000 Gift from P. Davies Phosphorylated Tau (pS422) IgG rabbit polyclonal to tau phospho-Serine 422 1:1000 (1:50) Invitrogen # 03151 Phosphorylated Tau (AT8) IgG1 mouse monoclonal to tau phospo-Serine 202 and phospho-Threonine 205 1:1000 (1:50) Invitrogen # MN1020 Truncated Tau (TauC3) IgG1 mouse monoclonal to truncated tau at 421/422 residues 1:1000 (1:50) ThermoFisher # AHB0061 Truncated Tau (MN423) IgG1 mouse monoclonal to truncated tau at glutamic acid 391 1:5000 Gift from L. Binder Choline Acetyltransferase (ChAT) IgG goat polyclonal to human placental choline acetyltransferase 1:800 (1:50) Millipore #AB144P Tyrosine Hydroxylase (TH) IgG rabbit polyclonal to tyrosine hydroxylase 1:1000 Invitrogen #OPA1–04050 Neurofilament (SMI-34) IgG1 mouse monoclonal to phosphorylated neurofilament H 1:1000 BioLegend #8535503 Sirtuin 6 IgG rabbit polyclonal to amino acids 330–348 of human Sirt6 1:5000 Sigma #S4197 Open in a separate window IH, Immunohistochemistry; IF, Immunofluorescence List of antibodies To visualize the relationship between amyloid plaques and tau-containing neurites, additional cortical and striatal sections were dual immunolabeled using two different colored chromogens.

Techniques: Labeling, Marker

Journal: Acta neuropathologica

Article Title: Frontal cortex and striatal cellular and molecular pathobiology in individuals with Down syndrome with and without dementia

doi: 10.1007/s00401-019-01965-6

Figure Lengend Snippet: List of antibodies

Article Snippet: To validate custom-designed microarray expression data, additional FC sections from both groups were incubated with a rabbit polyclonal antibody against the human nuclear sirtuin 6 (SIRT 6; 1:5000, Sigma), after performing antigen retrieval using Dako target retrieval solution, pH 9 (DAKO, Carpinteria, CA) for 20 minutes in a steamer [ 96 ]. table ft1 table-wrap mode="anchored" t5 Table 2. caption a7 Antigen Antibody Dilution IH (IF) Company Catalog # APP/Aβ (6E10) IgG1 mouse monoclonal to residues 1–16 of Aβ 1:1000 BioLegend #803002 Aβ (MOAB2) IgG2b mouse monoclonal to Aβ 1:800 Gift from M. J. LaDu Aβ42 IgG rabbit polyclonal to Aβ42 1:1000 Millipore #AB5078 Conformational Tau (Alz50) IgM mouse monoclonal to tau residues 5–15, 312–322 1:1000 (1:50) Gift from P. Davies Conformational Tau (MC1) IgG1 mouse monoclonal to tau residues 5–15, 312–322 1:1000 Gift from P. Davies Phosphorylated Tau (pS422) IgG rabbit polyclonal to tau phospho-Serine 422 1:1000 (1:50) Invitrogen # 03151 Phosphorylated Tau (AT8) IgG1 mouse monoclonal to tau phospo-Serine 202 and phospho-Threonine 205 1:1000 (1:50) Invitrogen # MN1020 Truncated Tau (TauC3) IgG1 mouse monoclonal to truncated tau at 421/422 residues 1:1000 (1:50) ThermoFisher # AHB0061 Truncated Tau (MN423) IgG1 mouse monoclonal to truncated tau at glutamic acid 391 1:5000 Gift from L. Binder Choline Acetyltransferase (ChAT) IgG goat polyclonal to human placental choline acetyltransferase 1:800 (1:50) Millipore #AB144P Tyrosine Hydroxylase (TH) IgG rabbit polyclonal to tyrosine hydroxylase 1:1000 Invitrogen #OPA1–04050 Neurofilament (SMI-34) IgG1 mouse monoclonal to phosphorylated neurofilament H 1:1000 BioLegend #8535503 Sirtuin 6 IgG rabbit polyclonal to amino acids 330–348 of human Sirt6 1:5000 Sigma #S4197 Open in a separate window IH, Immunohistochemistry; IF, Immunofluorescence List of antibodies To visualize the relationship between amyloid plaques and tau-containing neurites, additional cortical and striatal sections were dual immunolabeled using two different colored chromogens.

Techniques:

Journal: Acta neuropathologica

Article Title: Frontal cortex and striatal cellular and molecular pathobiology in individuals with Down syndrome with and without dementia

doi: 10.1007/s00401-019-01965-6

Figure Lengend Snippet: a. Histogram showing a downregulation of mean mRNA expression levels of several cholinergic [acetylcholinesterase (Ache), butyrylcholinesterase (Bche), muscarinic receptor M2 (Chrm2), M4 (Chmr4), cholinergic receptor nicotinic alpha 2 (Chrna2), 3 (Chrna3), 4(Chrna4) and 7 (Chrna7) subunits] and monoaminergic [α1B adrenergic receptor (Adra1b), serotonin receptor 1B (Htr1b), 2C (Htr2c), 3A (Htr3a), 7 (Htr7), monoamine oxidase A (Maoa) and B (Maob), noradrenaline transporter (Slc6a2), serotonin transporter (Slc6a4) and tyrosine hydroxylase (Th)] neurotransmission related genes in frontal cortex pS422 positive NFTs in demented compared to non-demented subjects with DS (Mann-Whitney rank sum test, p<0.05). b. Histogram showing a downregulation of the mean values of the relative density of autoimmune regulator (Aire), FAM3B protein (Fam3b), homeobox 2 (Emx2) transcript factor, the autophagy protein 5 (Atg5), TNF type-1 (Tradd), protein p53 (Tp53), caspase 6 (Casp6), BCL2-associated X (Bax), several cyclin proteins [cyclin B1 (Ccnb1), cyclin D1 (Ccnd1) and cyclin D2 (Ccnd2)], G-proteins [(Rgs2), (Rgs3), (Rgs4), (Rgs9), (Rgs10)], adenylate cyclases [(Adcy1), (Adcy6)], proto-oncogene c-fos (Fos), Jun (Jun), protein fosB (Fosb), and transcript factor EB (Tfeb) mRNAs in frontal cortex pS422 positive NFTs in demented compared to non-demented subjects with DS, while transcript expression levels for the Sirt6 gene was upregulated in frontal cortex pS422 positive NFTs in demented compared to non-demented subjects with DS (Mann-Whitney rank sum test, p<0.05).

Article Snippet: To validate custom-designed microarray expression data, additional FC sections from both groups were incubated with a rabbit polyclonal antibody against the human nuclear sirtuin 6 (SIRT 6; 1:5000, Sigma), after performing antigen retrieval using Dako target retrieval solution, pH 9 (DAKO, Carpinteria, CA) for 20 minutes in a steamer [ 96 ]. table ft1 table-wrap mode="anchored" t5 Table 2. caption a7 Antigen Antibody Dilution IH (IF) Company Catalog # APP/Aβ (6E10) IgG1 mouse monoclonal to residues 1–16 of Aβ 1:1000 BioLegend #803002 Aβ (MOAB2) IgG2b mouse monoclonal to Aβ 1:800 Gift from M. J. LaDu Aβ42 IgG rabbit polyclonal to Aβ42 1:1000 Millipore #AB5078 Conformational Tau (Alz50) IgM mouse monoclonal to tau residues 5–15, 312–322 1:1000 (1:50) Gift from P. Davies Conformational Tau (MC1) IgG1 mouse monoclonal to tau residues 5–15, 312–322 1:1000 Gift from P. Davies Phosphorylated Tau (pS422) IgG rabbit polyclonal to tau phospho-Serine 422 1:1000 (1:50) Invitrogen # 03151 Phosphorylated Tau (AT8) IgG1 mouse monoclonal to tau phospo-Serine 202 and phospho-Threonine 205 1:1000 (1:50) Invitrogen # MN1020 Truncated Tau (TauC3) IgG1 mouse monoclonal to truncated tau at 421/422 residues 1:1000 (1:50) ThermoFisher # AHB0061 Truncated Tau (MN423) IgG1 mouse monoclonal to truncated tau at glutamic acid 391 1:5000 Gift from L. Binder Choline Acetyltransferase (ChAT) IgG goat polyclonal to human placental choline acetyltransferase 1:800 (1:50) Millipore #AB144P Tyrosine Hydroxylase (TH) IgG rabbit polyclonal to tyrosine hydroxylase 1:1000 Invitrogen #OPA1–04050 Neurofilament (SMI-34) IgG1 mouse monoclonal to phosphorylated neurofilament H 1:1000 BioLegend #8535503 Sirtuin 6 IgG rabbit polyclonal to amino acids 330–348 of human Sirt6 1:5000 Sigma #S4197 Open in a separate window IH, Immunohistochemistry; IF, Immunofluorescence List of antibodies To visualize the relationship between amyloid plaques and tau-containing neurites, additional cortical and striatal sections were dual immunolabeled using two different colored chromogens.

Techniques: Expressing, MANN-WHITNEY

Journal: Acta neuropathologica

Article Title: Frontal cortex and striatal cellular and molecular pathobiology in individuals with Down syndrome with and without dementia

doi: 10.1007/s00401-019-01965-6

Figure Lengend Snippet: List of antibodies

Article Snippet: To validate custom-designed microarray expression data, additional FC sections from both groups were incubated with a rabbit polyclonal antibody against the human nuclear sirtuin 6 (SIRT 6; 1:5000, Sigma), after performing antigen retrieval using Dako target retrieval solution, pH 9 (DAKO, Carpinteria, CA) for 20 minutes in a steamer [ 96 ]. table ft1 table-wrap mode="anchored" t5 Table 2. caption a7 Antigen Antibody Dilution IH (IF) Company Catalog # APP/Aβ (6E10) IgG1 mouse monoclonal to residues 1–16 of Aβ 1:1000 BioLegend #803002 Aβ (MOAB2) IgG2b mouse monoclonal to Aβ 1:800 Gift from M. J. LaDu Aβ42 IgG rabbit polyclonal to Aβ42 1:1000 Millipore #AB5078 Conformational Tau (Alz50) IgM mouse monoclonal to tau residues 5–15, 312–322 1:1000 (1:50) Gift from P. Davies Conformational Tau (MC1) IgG1 mouse monoclonal to tau residues 5–15, 312–322 1:1000 Gift from P. Davies Phosphorylated Tau (pS422) IgG rabbit polyclonal to tau phospho-Serine 422 1:1000 (1:50) Invitrogen # 03151 Phosphorylated Tau (AT8) IgG1 mouse monoclonal to tau phospo-Serine 202 and phospho-Threonine 205 1:1000 (1:50) Invitrogen # MN1020 Truncated Tau (TauC3) IgG1 mouse monoclonal to truncated tau at 421/422 residues 1:1000 (1:50) ThermoFisher # AHB0061 Truncated Tau (MN423) IgG1 mouse monoclonal to truncated tau at glutamic acid 391 1:5000 Gift from L. Binder Choline Acetyltransferase (ChAT) IgG goat polyclonal to human placental choline acetyltransferase 1:800 (1:50) Millipore #AB144P Tyrosine Hydroxylase (TH) IgG rabbit polyclonal to tyrosine hydroxylase 1:1000 Invitrogen #OPA1–04050 Neurofilament (SMI-34) IgG1 mouse monoclonal to phosphorylated neurofilament H 1:1000 BioLegend #8535503 Sirtuin 6 IgG rabbit polyclonal to amino acids 330–348 of human Sirt6 1:5000 Sigma #S4197 Open in a separate window IH, Immunohistochemistry; IF, Immunofluorescence List of antibodies To visualize the relationship between amyloid plaques and tau-containing neurites, additional cortical and striatal sections were dual immunolabeled using two different colored chromogens.

Techniques: