Journal: bioRxiv

Article Title: CD164 is an endolysomal host factor for entry of Clade A New World Arenaviruses

doi: 10.64898/2026.04.21.719929

Figure Lengend Snippet: (A) Coomassie-stained SDS-PAGE of soluble proteins used in . 2.5 µg of PNGaseF treated, nondenatured, or denatured purified protein. (B) Nonlinear fit of binding curves at pH 5.0 (blue) and 7.4 (grey) for PICV sGP1, PARV sGP1, FLEXV sGP1, and MACV sGP1 to 100 nM of immobilized sCD164. Curves represent the averaged final response at the end of association (n=3, independent experiments in triplicate). (C) Binding of 5 µM MACV sGP1 (maroon triangle) or 1 µg mouse anti-human TfR1 (CD71) antibody (cyan diamond) to 100 nM of immobilized hTfR1-Fc as a function of pH. Normalized to max binding at pH 8.0 (n=2, independent experiments in duplicate). (D) pH 7.4 binding curve of MACV sGP1 to hTfR1 (100 nM). Left: 1:1 Langmuir fit; middle: nonlinear fit; right: Scatchard plot. Binding affinity (K D ) calculated from 1:1 Langmuir fit, K D = 2.52 µM ± 0.006 µM. (n=2, independent experiments in duplicate). (E) Representative histogram of mean fluorescence intensity (MFI) from . Cells bound with 2.5 µM AF647-labeled PICV sGP1 at pH=6.0 (orange: CD164 KO +WT-CD164 PM ; purple: CD164 KO ) or pH 7.4 (red: CD164 KO +CD164 PM ; blue: CD164 KO ) (n=3, independent experiments in triplicate) (F) MFI (/10,000) of CD164 KO +WT-CD164 PM or CD164 KO cells from and S4E, labeled with 5 µM AF647-PICV sGP1 at pH 6.0 and then washed at pH 7.4 (hashed) or 6.0 (solid). (n=3, independent experiments in triplicate).

Article Snippet: For each binding assay, sCD164-Fc, hTfR1-Fc, sCRD-Fc, or mutant CD164, were first immobilized on GatorBio anti-human IgG Fc Gen II (HFCII) probes (GatorBio,160024) for 300 sec.

Techniques: Staining, SDS Page, Purification, Binding Assay, Fluorescence, Labeling

Journal: bioRxiv

Article Title: CD164 is an endolysomal host factor for entry of Clade A New World Arenaviruses

doi: 10.64898/2026.04.21.719929

Figure Lengend Snippet: (A) Reducing/denaturing or non-denaturing Coomassie of sCD164 mutant proteins . Asterisk (*) denotes denaturing. (B) Final binding response of 1 µg N6B6 anti-human CD164 antibody to 100nM immobilized sCD164 mutants. (C) Inhibition of chimeric VSV expressing indicated glycoproteins by anti-CD164 monoclonal antibody, N6B6. Cells were pretreated with N6B6 for 1 hour on ice, virus was then added at MOI=1. At 6 hours post-infection, cells were fixed and % GFP-positive cells was determined by flow cytometry. % GPF+ was normalized to HeLa cells infected with no inhibition (n=2). (D) Relative expression of CD164 in CD164 mutant addback cells. Top: area under the curve (CD164/actin) compared to WT; bottom: representative Western blot. (E) Representative histograms of total CD164 expression in CD164 mutant addback cells.

Article Snippet: For each binding assay, sCD164-Fc, hTfR1-Fc, sCRD-Fc, or mutant CD164, were first immobilized on GatorBio anti-human IgG Fc Gen II (HFCII) probes (GatorBio,160024) for 300 sec.

Techniques: Mutagenesis, Binding Assay, Inhibition, Expressing, Virus, Infection, Flow Cytometry, Western Blot

Journal: Journal of Translational Autoimmunity

Article Title: Anti-IgLON5 encephalitis is associated with anti-retinal immunological reactivity without retinal alteration

doi: 10.1016/j.jtauto.2026.100359

Figure Lengend Snippet: Indirect immunofluorescence on sections of monkey retina. The fluorescence observed is identified by a black arrow. (A) Patient 4 (IgG4); (B) Patient 1 (IgG4); (C) Patient 3 (IgG1); (D) Patient 5 (IgG4); (E) Patient 2 (IgG1); (F) Patient 1 (IgG4) after immunoadsorption of IgLON5 antibodies; (G) Control with macular edema (IgG1); (H) Control with anti-Hu encephalitis (IgG1); (I) Control with CAR syndrome (IgG4). The different layers of the retina are identified by their initials: pigment epithelium (pe), photoreceptor layer (pr), outer grain layer (og), outer plexiform layer (op), inner grain layer (ig), inner plexiform layer (ip), ganglion cell layer (gc), nerve fibre layer (nf).

Article Snippet: The reactivity of the patients’ sera and/or CSF against the retina was evaluated retrospectively with frozen samples (−80°, EXPLAINEUR biobank) through an indirect immunofluorescence technique using sections of monkey retina (Ref. FA1172-1005, Euroimmun), detected with an FITC-labelled secondary antibody anti-human IgAGM (Euroimmun conjugate) or directed against IgA (ref. F0204, DAKO), IgM (ref. F0203, DAKO), IgG1 (ref. 9052-02, Southern Biotech) and IgG4 (ref. 9200-02, Southern Biotech).

Techniques: Immunofluorescence, Fluorescence, Control

Journal: bioRxiv

Article Title: Anti-HIV Immunotoxin and Antibody-Drug Conjugate Display Both Common and Distinct Effects in Killing Target Cells

doi: 10.64898/2026.04.07.717054

Figure Lengend Snippet: A . The binding of anti-gp41 7B2 to H9/N:L4-3 cells was demonstrated by indirect immunofluorescence and flow cytometry. The gray histogram indicates binding in the presence of secondary FITC-conjugated goat anti-human IgG only, the blue histogram demonstrates fluorescence of cells first incubated with 7B2 followed by FITC-conjugated secondary antibody. B. Comparative cytotoxicity of 7B2-dgA and 7B2-PNU. Cells were incubated with the indicated concentration of CIC and CD4-IgG2 (500 ng/mL) for 72 hr. MTS dye reduction was measured during the final 3 hr of culture. C. The kinetics of apoptosis induction by the CICs was measured by the binding of fluorescent annexin V to cells at different times following treatment with CIC.

Article Snippet: Cells were washed twice in PBS and incubated with 2 μg /mL of secondary FITC-conjugated goat anti-human IgG (Novus Biologicals, Centennial CO USA) for 1 hr at room temperature in the dark, then washed a final time with PBS and resuspended in 2% paraformaldehyde.

Techniques: Binding Assay, Immunofluorescence, Flow Cytometry, Fluorescence, Incubation, Concentration Assay