Journal: bioRxiv

Article Title: An Advanced Humanized Systemic Lupus Erythematosus Model Enables Parallel Profiling of B Cell-Targeted Therapies

doi: 10.64898/2026.01.27.701893

Figure Lengend Snippet: (a) Phenotypic analysis of human immune cells in spleens from NCG HIS (n=5) and NCG-M HIS mice (n=6) . Frequencies and absolute numbers of CD19 + (B cells), CD3 + (T cells), CD19 − CD3 − (nonTB), CD19 − CD3 − HLA-DR + CD123 + (pDC) and CD19 − CD3 − HLA-DR + CD1c + (cDC) cells were summarized. (b) Representative plots of . (c-d) Immunofluorescence staining of human IgG (c) and mouse complement3(C3, d) deposition in glomerular area of kidneys from NCG-M HIS-Ctrl and NCG-M HIS-SLE group. Scare bar, 50 or 200μm.Data show mean ± SEM. Statistical analysis was performed using unpaired two-tailed t -test. The exact p values are shown.

Article Snippet: T cells collected from healthy donors were subjected to enrichment with magnetic separation using anti-CD3 microbeads (Miltenyi Biotec) and activation with T Cell TransAct (Miltenyi Biotec).

Techniques: Immunofluorescence, Staining, Two Tailed Test

Journal: bioRxiv

Article Title: An Advanced Humanized Systemic Lupus Erythematosus Model Enables Parallel Profiling of B Cell-Targeted Therapies

doi: 10.64898/2026.01.27.701893

Figure Lengend Snippet: A) NCG-M HIS mice were either topically treated with TLR7 agonist (R848-induced-lupus group, HIS-RIL, n=3 ) or intraperitoneal injection with 500μl pristane (Pristane-induced-lupus group, HIS-PIL, n=4 ) for 8 weeks, disease phenotypes were determined at the endpoint. B-C) Representative images(B) and summarized data(C) of anti-nuclear antibody in serum from two groups detected by IIF. Scale bar, 100μm. D) Serum anti-dsDNA antibody levels detected by ELISA. E) Disease associated cytokine levels in serum detected by Legendplex. F) Urea nitrogen and creatinine level in serum from two groups detected by biochemical analysis. G) Immunofluorescence staining of human IgM (left, whole section) and human IgG (mid, whole section and right, enlarged version) deposition in kidneys from HIS-PIL and HIS-RIL group. Scare bar, 1mm or 100μm. H) Phenotypic analysis of B cells in spleen and kidney tissues from HIS-PIL and HIS-RIL group. Frequencies of CD24 − CD38 hi (plasmablast), CD38 − CD27 + (memory) and CD86 + (activated) cells were summarized. I) Phenotypic analysis of T cells in spleen and kidney tissues from HIS-PIL and HIS-RIL group. Frequencies of CD45RA + CCR7 + (naive), CD45RA + CCR7 − (Temra), CD45RA − CCR7 + (Tcm) and CD45RA − CCR7 − (Tem) cells were summarized. J) Phenotypic analysis of human immune cells in skin tissues from HIS-PIL and HIS-RIL group. Frequencies of CD3 + (T cells), CD19 + (B cells) and CD3 − CD19 − (nonTB) cells were summarized. Data show mean ± SEM. Statistical analysis was performed using unpaired two-tailed t -test. The exact p values are shown.

Article Snippet: T cells collected from healthy donors were subjected to enrichment with magnetic separation using anti-CD3 microbeads (Miltenyi Biotec) and activation with T Cell TransAct (Miltenyi Biotec).

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Two Tailed Test

Journal: bioRxiv

Article Title: An Advanced Humanized Systemic Lupus Erythematosus Model Enables Parallel Profiling of B Cell-Targeted Therapies

doi: 10.64898/2026.01.27.701893

Figure Lengend Snippet: (a) The schematic diagram of UCAR-T cells preparation. (b)Allogeneic CAR-T cells were cocultured with PBMCs for 24 hours at indicated effector (UCAR-T cells) to target (B cells) ratio. Frequencies of B cells and calculated specific killing efficiency were summarized. (c) PBMCs were cultured in complete medium for 24 hours with different concentrations of CD3-CD19 bispecific antibody. Frequencies of B cells and calculated specific killing efficiency were summarized. (d) Flowcytometry analysis of absolute numbers of B cells in peripheral blood at indicated time points. (e-f) Representative plots(e) and summarized data(f) of frequencies of transitional B cells (CD24 hi CD38 hi ) in peripheral blood at indicated time points (related to ). (g) The schematic diagram of allogeneic CAR-T cells transfer and analysis. Five million UCAR-T cells were transferred to HIS-NCG-M mice and tissue infiltrated cells were analyzed one week later. (h-j) Phenotypic analysis of UCAR-T cells in different tissues. (h)Representative plots showing B cells depletion, UCAR-T cells infiltration and the activation (CD69 + ) and exhaustion (PD-1 + ) state of UCAR-T cells in peripheral blood(up) or bone marrow tissues(down). (i-j) Absolute numbers of UCAR-T cells(i) and frequencies of CD69 + and PD-1 + cells(j) were summarized. Data show mean ± SEM.

Article Snippet: T cells collected from healthy donors were subjected to enrichment with magnetic separation using anti-CD3 microbeads (Miltenyi Biotec) and activation with T Cell TransAct (Miltenyi Biotec).

Techniques: Cell Culture, Activation Assay

Journal: bioRxiv

Article Title: An Advanced Humanized Systemic Lupus Erythematosus Model Enables Parallel Profiling of B Cell-Targeted Therapies

doi: 10.64898/2026.01.27.701893

Figure Lengend Snippet: A) Quality control of allogeneic CAR-T(UCAR-T) cells. Representative plots of TCR, HLA-A2 and CAR expression on UCAR-T cells were shown. B) The schematic diagram of UCAR-T cells and Blinatumomab in vivo application. Uninduced NCG-M HIS mice (HIS-Ctrl) were treated with either one million UCAR-T cells once( n=3 ) or T cell engager Blinatumomab at D0,1,2,4 and 6( n=3 ). The number of B cells in peripheral blood was monitored. C) Flowcytometry analysis of numbers of UCAR-T cells in peripheral blood at indicated time points. D) Flowcytometry analysis of frequencies of B cells in peripheral blood at indicated time points. E) Flowcytometry analysis of frequencies of mature B cells (CD24 int CD38 int ) in peripheral blood at indicated time points. F) The schematic diagram of UCAR-T cells and Blinatumomab treatment in NCG-M HIS-SLE mice. NCG-M HIS-SLE mice induced with TLR7 agonist were treated with vehicle (Vehicle group, n=3 ), allogeneic CAR-T cells (UCAR-T group, n=5 ) or CD3-CD19 bispecific antibody (Blinatumomab group, n=5 ) for four weeks. Disease phenotypes were determined at the end point. G) Anti-dsDNA and anti-smith antibody levels in serum from three groups detected by ELISA. H) Disease associated cytokine levels in serum from three groups detected by Legendplex. I-J) Kidney function of three groups. Urea nitrogen levels in serum(I)and protein levels in urine(J) were summarized. K) Phenotypic analysis of B cells in spleen tissues from Vehicle, Blinatumomab and UCAR-T group. Frequencies of CD24 hi CD38 hi (transitional B), IgD + CD27 − (naïve B), CD38 − CD27 + (memory B) and CD24 − CD38 hi (plasmablast) cells were summarized. L) Phenotypic analysis of T cells in spleen tissues from Vehicle, Blinatumomab and UCAR-T group. Frequencies of CD45RA + CCR7 + (naive), CD45RA + CCR7 − (Temra), CD45RA − CCR7 + (Tcm) and CD45RA − CCR7 − (Tem) cells were summarized. M) Flowcytometry analysis of frequencies of CD69 + (activated) T cells in spleen, bone marrow and kidney tissues from three groups. N-P) Tissue pathology detected by HE staining and Masson Trichrome staining. (N) HE staining of lung tissues from three groups. Scare bar, 500μm. (O) HE staining of liver tissues from three groups. Arrows indicate the immune cells infiltrated area. Scare bar, 200μm. (P) Masson Trichrome staining of interstitial area of kidneys from three groups. Arrows indicate the fibrotic areas. Scare bar, 100μm. Q-R) Safety evaluation of the two therapies. (Q) Monitored body weight over the whole treatment period. (R) Cytokines in serum from three groups 24 hours after first injection. Data show mean ± SEM. Statistical analysis was performed using one-way ANOVA with Tukey multiple-comparison test. The exact p values are shown.

Article Snippet: T cells collected from healthy donors were subjected to enrichment with magnetic separation using anti-CD3 microbeads (Miltenyi Biotec) and activation with T Cell TransAct (Miltenyi Biotec).

Techniques: Control, Expressing, In Vivo, Enzyme-linked Immunosorbent Assay, Staining, Injection, Comparison