Journal: Journal of Neuroinflammation

Article Title: CD22 modulation alleviates amyloid β-induced neuroinflammation

doi: 10.1186/s12974-025-03361-2

Figure Lengend Snippet: CD22 promotes neuroinflammation via microglia. A Schematic diagram of sCD22 i.c.v. injection into wildtype C57BL/6 mice. B Volcano plot of sCD22-treated mouse cortex after 3 days treatment. N = 3. C-D Gene ontology analysis of sCD22-treated mice (3 days treatment) in KEGG pathway ( C ) and Biological function ( D ). E Volcano plot of sCD22-treated mouse cortex after 7 days treatment. N = 4. F Gene ontology analysis of sCD22-treated mice (7 days treatment) in Biological function. G GSEA showing enrichment of IL6/JAK/STAT3, TNFα Signaling via NFκB, and Cholesterol Homeostasis of mouse cortex after sCD22 7 days treatment relative to PBS group. H Representative image and quantitation showing the effect of sCD22 on IbaI and GFAP expression in mouse cortex. IbaI: Student t -test, ** P < 0.01; GFAP: ns: not significant. N = 6. I Schematic diagram of MDMi differentiation and sCD22 treatment. J Principal component analysis showing sCD22 treated MDMi versus control MDMi. N = 4. K Volcano plot of sCD22-treated MDMi. L Gene ontology analysis of sCD22-treated MDMi in Biological function. M Gene ontology analysis of sCD22-treated MDMi in KEGG

Article Snippet: All other materials included RPMI-1640 (Gibco, A10491-01), Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, 11995-040), FBS (Gibco, A5256701), Pen/strep (Gibco, 15140-122), IL-34 (Sino Biological, 10948-H08S), GM-CSF (Sino Biological, 10015-HNAH), Geltrex (Gibco, A14133-02), human CD22 ECD (CD22 FL) (Sino Biological, 11958-H08H), human CD22 a.a. 176–687 (CD22 δ1) (Sino Biological, 11958-H08H1), mouse CD22 ECD (Sino Biological, 51177-M08H), Tanzisertib (Selleckchem, S8490), Perifosine (Selleckchem, S1037), Ravoxertinib (Selleckchem, S7554), SB856553 (Selleckchem, S7215).

Techniques: Injection, Quantitation Assay, Expressing, Control

Journal: Journal of Neuroinflammation

Article Title: CD22 modulation alleviates amyloid β-induced neuroinflammation

doi: 10.1186/s12974-025-03361-2

Figure Lengend Snippet: sCD22 promotes microglial neuroinflammation via MAPK-signaling pathway and in a sialic acid-dependent manner. A Immunostaining with anti-IbaI antibody to examine microglia activation after sCD22 treatment in MDMi. Student t -test, **** P < 0.0001, ns: not significant. N = 26–27, from 3 independent experiments. B-D Effect of sCD22 on viability of MDMi ( B ), HMC-3 ( C ) and BV-2 cells ( D ). E Representative and quantitation of western blot examining ERK1/2 and p38 phosphorylation in sCD22-treated MDMi. p38: One-way ANOVA, F = 16.01, P = 0.001, Tukey post hoc test ** P < 0.01; ERK1/2: One-way ANOVA, F = 7.38, P = 0.0108, Tukey post hoc test * P < 0.05. F Effect of ERK1/2 inhibitor (Ravoxertinib) and p38 inhibitor (SB856553) on sCD22-mediated TNFα, IL-6 & CCL3 release. TNFα: One-way ANOVA, F = 16.09, Tukey post hoc test **** P < 0.0001; IL-6: One-way ANOVA, F = 4.917, Tukey post hoc test * P < 0.05, ** P < 0.01; CCL3: One-way ANOVA, F = 6.672, Tukey post hoc test ** P < 0.01. N = 4–5. G Effect of pan JNK inhibitor (Tanzisertib) and Akt inhibitor (Perifosine) on sCD22-mediated CCL3 release. H Schematic diagram of sCD22 with complete extracellular domain (CD22-FL) and with D1-truncated (CD22-δ1). I Full length and D1-truncated sCD22 effect on TNFα, IL-6 & CCL3 release in MDMi. TNFα: One-way ANOVA, F = 7.847, Tukey post hoc test * P < 0.05, ** P < 0.01; IL-6: One-way ANOVA, F = 4.375, Tukey post hoc test * P < 0.05; CCL3: One-way ANOVA, F = 3.669, Tukey post hoc test * P < 0.05. ns: not significant. N = 5–6. J Effect of CHO-derived sCD22 on CCL3 release in MDMi. Student t -test, P = 0.83. N = 2. K Effect of HEK293-derived sCD22 and CHO-derived sCD22 on CCL3 release in THP-1. Two-way ANOVA, source of sCD22: F (1,4) = 124, P = 0.0007; CCL3 release: F (1,4) = 87.57, P = 0.0007. Tukey post hoc test, i < 0.001. ns: not significant. N = 2. All data are presented as mean ± SEM

Article Snippet: All other materials included RPMI-1640 (Gibco, A10491-01), Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, 11995-040), FBS (Gibco, A5256701), Pen/strep (Gibco, 15140-122), IL-34 (Sino Biological, 10948-H08S), GM-CSF (Sino Biological, 10015-HNAH), Geltrex (Gibco, A14133-02), human CD22 ECD (CD22 FL) (Sino Biological, 11958-H08H), human CD22 a.a. 176–687 (CD22 δ1) (Sino Biological, 11958-H08H1), mouse CD22 ECD (Sino Biological, 51177-M08H), Tanzisertib (Selleckchem, S8490), Perifosine (Selleckchem, S1037), Ravoxertinib (Selleckchem, S7554), SB856553 (Selleckchem, S7215).

Techniques: Immunostaining, Activation Assay, Quantitation Assay, Western Blot, Derivative Assay

Journal: Journal of Neuroinflammation

Article Title: CD22 modulation alleviates amyloid β-induced neuroinflammation

doi: 10.1186/s12974-025-03361-2

Figure Lengend Snippet: CD22 modulation by suciraslimab alleviates Aβ-induced neuroinflammation in human CD22 transgenic mice. A Schematic diagram of Aβ-induced neuroinflammation model in human CD22 transgenic mice. B Effect of suciraslimab on Aβ-injected model mice in Y-maze test. Alternation: One-way ANOVA, F = 4.724, P = 0.0196. Tukey post hoc test, * P < 0.05. Number of arm entry: One-way ANOVA, F = 0.07, P = 0.93. N = 8–9. C Volcano plot of suciraslimab-treated mouse cortex. N = 3. D Gene ontology analysis of suciraslimab-treated mouse cortex in Biological function. E Gene ontology analysis of suciraslimab-treated mouse cortex in molecular function. F Effect of suciraslimab on chemokine release in mouse brain of model mice. Student t -test, P value as stated in the figure. N = 3. All data are presented as mean ± SEM

Article Snippet: All other materials included RPMI-1640 (Gibco, A10491-01), Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, 11995-040), FBS (Gibco, A5256701), Pen/strep (Gibco, 15140-122), IL-34 (Sino Biological, 10948-H08S), GM-CSF (Sino Biological, 10015-HNAH), Geltrex (Gibco, A14133-02), human CD22 ECD (CD22 FL) (Sino Biological, 11958-H08H), human CD22 a.a. 176–687 (CD22 δ1) (Sino Biological, 11958-H08H1), mouse CD22 ECD (Sino Biological, 51177-M08H), Tanzisertib (Selleckchem, S8490), Perifosine (Selleckchem, S1037), Ravoxertinib (Selleckchem, S7554), SB856553 (Selleckchem, S7215).

Techniques: Transgenic Assay, Injection

Journal: Journal of Neuroinflammation

Article Title: CD22 modulation alleviates amyloid β-induced neuroinflammation

doi: 10.1186/s12974-025-03361-2

Figure Lengend Snippet: Suciraslimab suppresses Aβ-induced inflammation in microglia and human PBMC. A Effect of CD22 overexpression on Aβ-induced NFκB signaling in HEK293. Two-way ANOVA: CD22 expression, F (1,20) = 62.97, i < 0.0001; Aβ treatment, F (4,20) = 16.83, P < 0.0001. Tukey post hoc test, **** P < 0.0001. N = 3. B Effect of suciraslimab on Aβ-induced IL-1β release in MDMi. One-way ANOVA, F = 7.767, P = 0.004. Tuley post hoc test, * P < 0.05, ** P < 0.01. N = 6–7. C Immunofluorescent staining and quantitation of NLRP3 and ASC after Aβ and suciraslimab treatment in MDMi. NLRP3: One-way ANOVA, F = 10.09, P < 0.0001, Tukey post hoc test * P < 0.05, *** P < 0.001; ASC, One-way ANOVA, F = 19.10, P < 0.0001, Tukey post hoc test **** P < 0.0001. N = 6–15. D Effect of suciraslimab on Aβ-induced IL-1β release in human PBMC. One-way ANOVA, F = 6.833, P = 0.0052. Tukey post hoc test, ** P < 0.01, ns = not significant. N = 8. E Effect of suciraslimab on IFNγ + LPS-induced IL-23 and IL-12 release in human PBMC. IL-23: One-way ANOVA, F = 26.93, P = 0.0002. Tukey post hoc test, ** P < 0.01, *** P < 0.001; IL-12: One-way ANOVA, F = 10.21, P = 0.0008. Tukey post hoc test, * P < 0.05, *** P < 0.001. N = 4–8. F Effect of suciraslimab on TLR4 surface expression on monocyte upon IFNγ and LPS activation. Two-tailed paired Student’s t test, P = 0.0293, t = 3.322, df = 4. N = 5 G Effect of suciraslimab on α4 integrin surface expression on T cell of human PBMC. One-way ANOVA, F = 0.7059, P = 0.5131. N = 5. H Effect of suciraslimab on α4 integrin surface expression on B cell of human PBMC. One-way ANOVA, F = 66.02, P < 0.0001. Tukey’s post hoc test, * P < 0.05, **** P < 0.0001. N = 4–5. I Effect of suciraslimab on α4 integrin surface expression on T cell-depleted human PBMC. One-way ANOVA, F = 16.91, P = 0.0009. Tukey’s post hoc test, ** P < 0.01. N = 4. J Effect of suciraslimab on α4 integrin surface expression on monocyte-depleted human PBMC. One-way ANOVA, F = 8.565, P = 0.0083. Tukey’s post hoc test, IgG1 vs. αCD22 Ab, P = 0.1748. N = 4. All data are presented as mean ± SEM

Article Snippet: All other materials included RPMI-1640 (Gibco, A10491-01), Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, 11995-040), FBS (Gibco, A5256701), Pen/strep (Gibco, 15140-122), IL-34 (Sino Biological, 10948-H08S), GM-CSF (Sino Biological, 10015-HNAH), Geltrex (Gibco, A14133-02), human CD22 ECD (CD22 FL) (Sino Biological, 11958-H08H), human CD22 a.a. 176–687 (CD22 δ1) (Sino Biological, 11958-H08H1), mouse CD22 ECD (Sino Biological, 51177-M08H), Tanzisertib (Selleckchem, S8490), Perifosine (Selleckchem, S1037), Ravoxertinib (Selleckchem, S7554), SB856553 (Selleckchem, S7215).

Techniques: Over Expression, Expressing, Staining, Quantitation Assay, Activation Assay, Two Tailed Test

Journal: Journal of Neuroinflammation

Article Title: CD22 modulation alleviates amyloid β-induced neuroinflammation

doi: 10.1186/s12974-025-03361-2

Figure Lengend Snippet: Suciraslimab promotes Aβ phagocytosis. A BLI analysis of mouse CD22-Aβ interaction. Association: 600s; Dissociation: 600s. B BLI analysis of human CD22-Aβ interaction. Association: 600s; Dissociation: 600s. C Immunofluorescent staining and quantitation of FITC-Aβ on HEK293 and HEK293-hCD22 cells. Student t -test, ** P < 0.01. N = 20–21, from 3 independent experiments. D Representative image and quantitation of Proximity-ligation assay of CD22-Aβ complex in HMC-3. Student’s t -test, **** P < 0.0001. N = 41, from 3 independent experiments. E Structural alignment of mouse CD22 and human CD22. The structures of both mouse and human CD22 extracellular domain were generated with Alphafold2. Pairwise structural alignment score (TM-score) higher than 0.5 assumes generally proteins aligned of the same fold. F Surface CD22 expression in HMC-3 after suciraslimab treatment. One-way ANOVA, F = 2.892, P = 0.0139. Tukey post hoc test, * P < 0.05. N = 76–83. G Surface suciraslimab binding on HMC-3. One-way ANOVA, F = 125, P < 0.0001. Tukey post hoc test, ** P = 0.002. N = 3. H Effect of suciraslimab on FITC-Aβ phagocytosis in HMC-3. One-way ANOVA, F = 43.92, P < 0.0001. Tukey post hoc test, * P = 0.046, ** P = 0.0018, **** P < 0.0001. N = 3. I Effect of suciraslimab on FITC-Aβ phagocytosis in PMA-differentiated MO3.13. Two-tailed Student’s t test, P = 0.0477, t = 2.482, df = 6

Article Snippet: All other materials included RPMI-1640 (Gibco, A10491-01), Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, 11995-040), FBS (Gibco, A5256701), Pen/strep (Gibco, 15140-122), IL-34 (Sino Biological, 10948-H08S), GM-CSF (Sino Biological, 10015-HNAH), Geltrex (Gibco, A14133-02), human CD22 ECD (CD22 FL) (Sino Biological, 11958-H08H), human CD22 a.a. 176–687 (CD22 δ1) (Sino Biological, 11958-H08H1), mouse CD22 ECD (Sino Biological, 51177-M08H), Tanzisertib (Selleckchem, S8490), Perifosine (Selleckchem, S1037), Ravoxertinib (Selleckchem, S7554), SB856553 (Selleckchem, S7215).

Techniques: Staining, Quantitation Assay, Proximity Ligation Assay, Generated, Expressing, Binding Assay, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Engineering the fragment crystallizable (Fc) region of human IgG1 multimers and monomers to fine-tune interactions with sialic acid-dependent receptors

doi: 10.1074/jbc.M117.795047

Figure Lengend Snippet: Binding of IgG1-Fc variants to glycan receptors. A , mutants lacking the Asn-297 glycan are severely restricted in their capacity to bind DC-SIGN by ELISA. The addition of an N -linked sugar at position 221 results in proteins with a reduced capacity to bind DC-SIGN compared with their equivalent variants in which Asn-221 is absent. B , the hypersialylated D221N mutants bind Siglec-1. No binding was observed with the N297A/N563A glycan-deficient mutant ( error bars represent standard deviations around the mean value, n = 2 independent experiments).

Article Snippet: The same ELISA protocol used to detect DC-SIGN binding was used for human Siglec-1, Siglec-4, and Siglec-3 (Sino Biologicals).

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Mutagenesis

Journal: The Journal of Biological Chemistry

Article Title: Engineering the fragment crystallizable (Fc) region of human IgG1 multimers and monomers to fine-tune interactions with sialic acid-dependent receptors

doi: 10.1074/jbc.M117.795047

Figure Lengend Snippet: Model showing the contribution of different N -linked glycan and cysteine residues on Fc stoichiometry. The presence of Cys-575 allows optimal disulfide bonding between tail pieces of monomeric-Fcs. The tail piece glycan Asn-563 controls the number of monomeric tails that fit into the central corona (five to six in the case of hexa-Fc) while still allowing Cys-309 interdisulfide bridge formation. Cys-575 allows disulfide bonding between tail pieces of different monomers, but the absence of the Asn-563 glycan (the N563A mutant) allows many more tail pieces (up to twelve in the case of dodecamers) to fit into the central corona while still allowing disulfide bond formation through Cys-309 and/or Cys-575. The absence of Cys-575 prevents disulfide bonding between tail pieces, thereby generating sialylated monomers at Asn-563. The additional Asn-563 tail piece glycan in these monomers must explain the increased binding seen to Siglec-1 ( , A and B , and inset in this figure). The bulkier Asn-563 glycan with its predicted overall negative charge may lead to repulsion between two monomers, thus preventing disulfide bond formation between two Cys-309 residues in each monomeric Fc. The loss of both Asn-563 and Cys-575 (the N563A/C575A mutant) means that the observed laddered multimers must arise through Cys-309–mediated disulfide bonding in the Cγ2 domain. The presence of monomers, dimers, trimers, tetramers, pentamers, hexamers, and other intermediates in this mutant ( C ) suggests that these structures arise through a different mechanism, most likely via the sequential addition of 25-kDa half-mer Fc units at Cys-309. The lack of observable ladders with the L448STOP mutant implies that other amino acids in the tail piece are involved in bringing about monomer interactions that then facilitate disulfide bonding through either Cys-309 and/or Cys-575. Monomers with glycans located at both the N- and C-terminal ends of the Fc (Asn-221 and Asn-563) may allow for binding to receptors in cis as shown ( inset ).

Article Snippet: The same ELISA protocol used to detect DC-SIGN binding was used for human Siglec-1, Siglec-4, and Siglec-3 (Sino Biologicals).

Techniques: Mutagenesis, Binding Assay

Journal: The Journal of Biological Chemistry

Article Title: Engineering the fragment crystallizable (Fc) region of human IgG1 multimers and monomers to fine-tune interactions with sialic acid-dependent receptors

doi: 10.1074/jbc.M117.795047

Figure Lengend Snippet: Binding of monomeric IgG1-Fc glycan variants to sialic acid-binding immunoglobulin-type lectins (Siglecs) with specificity for α2,3-linked sialic acid. A , the C575A monomer binds Siglec-1. B , the D221N/C575A monomer binds Siglec-1 and Siglec-4. ELISA as described under “Experimental procedures” with receptors coated down at 2 μg/ml and Fc-fragments at 20 μg/ml in TMS buffer ( error bars represent standard deviations around the mean value, n = 2 independent experiments).

Article Snippet: The same ELISA protocol used to detect DC-SIGN binding was used for human Siglec-1, Siglec-4, and Siglec-3 (Sino Biologicals).

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay