Journal: Cancer research

Article Title: A Re-evaluation of CD22 Expression by Human Lung Cancer

doi: 10.1158/0008-5472.CAN-13-1436

Figure Lengend Snippet: Relative fold expression of CD22 mRNA as compared to GAPDH in the cell lines included in the study, as measured by qRT- PCR a

Article Snippet: Gene specific TaqMan probes (Applied Biosystems, Foster City, CA) were utilized for quantitative analyses of mRNA transcript levels (CD22 Hs00233533_m1, GAPDH 02758991_g1).

Techniques: Expressing, Quantitative RT-PCR, Gene Expression

Journal: Cancer research

Article Title: A Re-evaluation of CD22 Expression by Human Lung Cancer

doi: 10.1158/0008-5472.CAN-13-1436

Figure Lengend Snippet: Representative PCR amplification of three distinct CD22 transcript regions using cDNA generated from the designated cell lines (1- Daudi; 2 - Jurkat; 3 - HCC827; 4 - H1355; 5 - H1975; 6 - A549; 7 - Calu-1; 8 - H1650; 9 - H727; 10 - dH2O). Daudi and Jurkat cells served as positive and negative controls, respectively. After 30 cycles of amplification, faint bands corresponding to CD22 transcripts were observed for Calu-1, H727, HCC827, H1650 and H1975 relative to the control (Daudi cells). Similar results were obtained in three or more experiments.

Article Snippet: Gene specific TaqMan probes (Applied Biosystems, Foster City, CA) were utilized for quantitative analyses of mRNA transcript levels (CD22 Hs00233533_m1, GAPDH 02758991_g1).

Techniques: Amplification, Generated, Control

Journal: Cancer research

Article Title: A Re-evaluation of CD22 Expression by Human Lung Cancer

doi: 10.1158/0008-5472.CAN-13-1436

Figure Lengend Snippet: A. Flow cytometric analysis. One million cells from each cell line were incubated with 25 µg/mL of either a mouse isotype control antibody (MPC-11) or mouse antihuman CD22 (clone HB22.7) MAb. After washing out the unbound primary antibody, a FITC-labeled secondary goat anti-human IgG antibody was used. Cells were analyzed in FL-1 using a BD FACSCalibur. Red line – cells without antibodies; green line – cells plus the isotype control antibody and secondary antibody; blue line – cells plus the anti-CD22 antibody and secondary antibody. This is one representative out of 4–6 independent experiments. B. WB analysis. Equal amounts (25 µg) of protein from cell lysates were subjected to 8% SDS-PAGE followed by a WB with a rabbit anti-human CD22 antibody (H-221 clone). Upper panel - CD22 expression; lower panel - β-actin expression (loading control): 1 – Daudi; 2 - no loading; 3 - Calu-1; 4 - H1975; 5 - H1650; 6- H1355; 7 - H1299; 8 - H1184; 9 - HCC827; 10 - H727; 11 - A549; 12 - H460; 13 - no loading; 14 – molecular weight marker for 140 kDa. This is one representative out of 3–5 independent experiments.

Article Snippet: Gene specific TaqMan probes (Applied Biosystems, Foster City, CA) were utilized for quantitative analyses of mRNA transcript levels (CD22 Hs00233533_m1, GAPDH 02758991_g1).

Techniques: Incubation, Control, Labeling, SDS Page, Expressing, Molecular Weight, Marker

Journal: Cancer research

Article Title: A Re-evaluation of CD22 Expression by Human Lung Cancer

doi: 10.1158/0008-5472.CAN-13-1436

Figure Lengend Snippet: Cell lines were cultured in quadruplicate in 96-well plates at 2 × 105 cells/mL in a volume of 100 µL complete medium. The following day, mouse anti-human CD22 MAbs (HB22.7 or RFB4) were added in a final volume of 200 µL at a final concentration ranging from 5 to 50 µg/mL (3.33 × 10−8 to 3.33 × 10−7 M). Both ITs [RFB4 (anti-CD22)-dgA and RFT5 (anti CD25)-dgA] were added in a final volume of 200 µL at a final molar concentration ranging from 1 × 10−10 to 1 × 10−13 M. Cells were incubated for 72 hours and cell viability was calculated by using CellTiter 96® AQueous One Solution [blank columns - no treatment, media only; horizontally hatched columns - HB22-7 at 3.33 × 10−7 M; checkered columns - RFB4 at 3.33 × 10−7 M; black columns - RFB4 (anti-CD22)-dgA at 1.0 × 10−11 M; light gray columns - RFT5 (anti-CD25)-dgA at 1.0 × 10−11 M]. The figure depicts the average ± SD of cell viability from three independent experiments.

Article Snippet: Gene specific TaqMan probes (Applied Biosystems, Foster City, CA) were utilized for quantitative analyses of mRNA transcript levels (CD22 Hs00233533_m1, GAPDH 02758991_g1).

Techniques: Cell Culture, Concentration Assay, Incubation

Journal: Cancer research

Article Title: A Re-evaluation of CD22 Expression by Human Lung Cancer

doi: 10.1158/0008-5472.CAN-13-1436

Figure Lengend Snippet: Immunostaining performed on serial sections of lung squamous cell carcinoma shows pan cytokeratin 5/6/18 positive cancer cells (A) and CD22 positive lymphocytes (B). Immunostaining performed on serial sections of lung adenocarcinoma shows thyroid transcription factor-1 positive cancer cells (C) and CD22 positive lymphocytes (D). Scale bars = 40 µm.

Article Snippet: Gene specific TaqMan probes (Applied Biosystems, Foster City, CA) were utilized for quantitative analyses of mRNA transcript levels (CD22 Hs00233533_m1, GAPDH 02758991_g1).

Techniques: Immunostaining

Journal: PLoS ONE

Article Title: Ovariectomy and Subsequent Treatment with Estrogen Receptor Agonists Tune the Innate Immune System of the Hippocampus in Middle-Aged Female Rats

doi: 10.1371/journal.pone.0088540

Figure Lengend Snippet: Expression of macrophage-associated genes and inhibitory neuronal ligands in the hippocampus of middle-aged female rats.

Article Snippet: Cd22 , Rn01457837_m1 , 10.54.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Ovariectomy and Subsequent Treatment with Estrogen Receptor Agonists Tune the Innate Immune System of the Hippocampus in Middle-Aged Female Rats

doi: 10.1371/journal.pone.0088540

Figure Lengend Snippet: Expression of Cd22 (A), Sema3a (B) and Cd200 (C) in the hippocampus was measured by real-time PCR. Error bars show SD of five samples for each group. There were statistically significant treatment effects in the case of Cd22 and Sema3a (p value of the ANOVA was smaller than 0.001 in both cases, p = 0.328 for Cd200). Turquoise, black and red asterisks indicate significant group differences compared to M, M/OVX, M/OVX+E2 animals, respectively, by Newman-Keuls post hoc test. * corresponds to 0.01

Article Snippet: Cd22 , Rn01457837_m1 , 10.54.

Techniques: Expressing, Real-time Polymerase Chain Reaction

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

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: The glucocorticoids prednisone and dexamethasone differentially modulate T cell function in response to anti-PD-1 and anti-CTLA-4 immune checkpoint blockade

doi: 10.1007/s00262-020-02555-2

Figure Lengend Snippet: Dexamethasone treatment impacts the expression of LAG-3 by CD8+ T cells. PBMCs were treated with the indicated concentrations of dexamethasone (dexa), anti-PD-1 (pembrolizumab (pem)), a-CTLA-4 (ipilimumab (ipi)) alone or in combination and stimulated with SEB for 72 h. a Representative FACS plots showing expression of LAG-3 by gated CD8+ T cells in response to dexamethasone treatment and SEB stimulation. b Cumulative data of LAG-3 expression by CD8+ T cells. Data from four independent experiments shown. Error bars represent mean ± SEM. P values obtained by ordinary one-way ANOVA followed by Dunnett’s multiple comparisons test. Unstim, unstimulated cells. c Plots showing phosphorylation of the SHP-2 tyrosine phosphatase Y542 by PD-1 + Jurkat cells stimulated with CHO cells which constitutively express PD-L + and a TCR activator. d PD-1negJurkat cells were pre-treated with the indicated steroids or anti-PD-1 (pembrolizumab) alone or in combination for 48 h. Jurkat cells were co-cultured with CHO cells for 3 h at a ratio of 10:1. Cell lysates were collected followed by measurement of Y542 phosphorylation by ELISA. e PBMCs were pre-treated with the indicated steroids or anti-PD-1 (pembrolizumab) alone or in combination for 3 h. Cell lysates were collected followed by measurement of Y542 phosphorylation by ELISA. Data from four experiments shown. Error bars represent mean ± SD. P values obtained by ordinary one-way ANOVA followed by Dunnett’s multiple comparisons test

Article Snippet: Recombinant Jurkat T cells expressing firefly luciferase gene under the control of NFAT with constitutive expression of PD-1 and Chinese Hamster Ovary (CHO) cells constitutively expressing PD-L1 and an engineered TCR activator were purchased from BPS Bioscience.

Techniques: Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Nature medicine

Article Title: Antigen-independent activation enhances the efficacy of 41BB co-stimulated CD22 CAR T cells

doi: 10.1038/s41591-021-01326-5

Figure Lengend Snippet: a, Pipeline for development and evaluation of new CD22-f2-short CAR. b, Affinity and size of purified CD22-f2-long and short scFvs. c, Expression of CD22 CARs on primary T cells. d, Measurement of secreted IFNγ by CD22-engineered T cells after 24h exposure to CD22+ target cells. e, Progression of Nalm6 disease burden in xenograft mice treated with CD22-f2-short and long T cells (Representative of 4 replicate experiments, n=4–7 mice per condition; see Supplementary Figure 5 for individual animal responses and Supplementary Figure 6 for experimental replicates). f, Survival of Nalm6-bearing xenograft mice after treatment with m971 or CD22-f2 CAR T cells. Data are presented as mean values +/− standard error of the mean (S.E.M.) Statistics reflect differences between CAR22-short and long T cells.

Article Snippet: 40 All animal studies were approved and supervised by the University of Pennsylvania Institutional Animal Care and Use Committee (IACUC). scFv design and optimization: To identify novel binders to human CD22 extracellular domain, three rounds of panning were done against recombinant human CD22 (R&D systems, Cat. # 1968-SL-050) using a fully human derived scFv phage library derived internally.

Techniques: Purification, Expressing

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: Free radical biology & medicine

Article Title: Extracellular vesicles released by ALL patients contain HNE-adducted proteins: Implications of collateral damage.

doi: 10.1016/j.freeradbiomed.2024.12.006

Figure Lengend Snippet: Fig. 3. Immunoblotting of protein markers in EV lysates. For all graphs in this figure, individual fold change was measured, followed by averaging the overall fold change across the different collection time points. Quantification of CD22 (A) and CD19 (B) as cell surface markers of leukemia. A significant increase of CD22 was detected in the EVs at consolidation day 15 compared to induction day 29 (p < 0.05). (C) A significant decrease in GFAP expression was observed in the EV lysates at consolidation days 1 and 8 compared to induction day 29 (p < 0.05). (D) An insignificant decline in the neuronal marker, NeuN, in the EVs was observed during consolidation phase compared to pre-treatment or induction day 29. (E) A statistically significant decrease in BDNF was observed in the EV lysates during both induction and consolidation phase collection points compared to pre-treatment (p < 0.05). *denotes p < 0.05 when compared to pre-treatment and # denotes p < 0.05 when compared to induction day 29. N.S. = No statistically significant difference.

Article Snippet: The primary antibodies and their dilutions used were: flotillin-1 (Flot-1) (1:20) from Bioss (catalog #: BS7798R), HSC70 (1:20) from Santa Cruz (catalog #: sc-7298), CD63 (1:10) from Santa Cruz (catalog #: sc-5275), ApoA1 (1:50) from Cell Signaling (catalog #: 3350), CD22 (1:50) from ProteinTech (catalog #: 66103-1-Ig) CD19 (1:10) from Abcam (catalog #: ab227019), GFAP (1:20) from Aviva Systems (catalog #: OAEB01041), BDNF (1:50) from Abcam (catalog #: ab10505), and NeuN from Novus (catalog #: NBP192716).

Techniques: Western Blot, Expressing, Marker