cd152 pevio770  (Miltenyi Biotec)

Journal: bioRxiv

Article Title: Structural basis of CD28 and CTLA-4 interactions with CD80, CD86, and the CD80-PD-L1 heterodimer on artificial and cellular membranes

doi: 10.64898/2026.03.30.715283

Figure Lengend Snippet: (A) Schematic of the lipid monolayer experiment. An octa-histidine tag was fused to the C-terminus of the CD86 ectodomain, enabling tethering of the CTLA-4-CD86 complex to the Ni 2+ -NTA lipid monolayer. “H” denotes the octa-histidine tag, and Ni 2+ -NTA lipids are shown in green. (B) Representative cryo-EM micrograph, with 2D class averages shown below. (C) Cryo-EM density map of the CTLA-4-CD86 complex. CTLA-4 is shown in dark and light red, whereas CD86 is shown in dark and light blue. (D) Overall structure of the CTLA-4-CD86 complex. (E) Close-up view of the CTLA-4-CD86 heterodimerization interface. The region outlined by the dashed pink box in (D) is enlarged.

Article Snippet: The gene encoding full-length CTLA-4 with an ALFA affinity tag was synthesized (Twist Bioscience) and cloned into the pEG BacMam transfer vector (Table S1) .

Techniques: Cryo-EM Sample Prep

Journal: bioRxiv

Article Title: Structural basis of CD28 and CTLA-4 interactions with CD80, CD86, and the CD80-PD-L1 heterodimer on artificial and cellular membranes

doi: 10.64898/2026.03.30.715283

Figure Lengend Snippet: (A) Schematic of the lipid monolayer experiment. An octa-histidine tag was fused to the C-terminus of the CD80 ectodomain, enabling tethering of the CTLA-4-CD80 complex to the Ni 2+ -NTA lipid monolayer. “H” denotes the octa-histidine tag, and Ni 2+ -NTA lipids are shown in green. (B) Representative cryo-EM micrograph, with 2D class averages shown below. (C) Cryo-EM density map of the CTLA-4-CD80 linear cluster, shown in grey. Refined atomic models of the CTLA-4-CD80 complex are fitted into the map. CTLA-4 is shown in red, whereas CD80 is shown in purple. (D) Three-dimensional refined cryo-EM map of the CTLA-4-CD80 linear cluster. (E) Overall structure of the CTLA-4-CD80 complex within the linear cluster. The IgV and IgC domains of CD80 are labeled. (F) Close-up view of the CTLA-4-CD80 heterodimerization interface. The region outlined by the dashed pink box in (E) is enlarged.

Article Snippet: The gene encoding full-length CTLA-4 with an ALFA affinity tag was synthesized (Twist Bioscience) and cloned into the pEG BacMam transfer vector (Table S1) .

Techniques: Cryo-EM Sample Prep, Labeling

Journal: bioRxiv

Article Title: Structural basis of CD28 and CTLA-4 interactions with CD80, CD86, and the CD80-PD-L1 heterodimer on artificial and cellular membranes

doi: 10.64898/2026.03.30.715283

Figure Lengend Snippet: (A) Representative cryo-EM micrograph of the CTLA-4-CD80 lattice. The region outlined by the red box is enlarged in the right panel. (B) 2D class averages of the CTLA-4-CD80 linear clusters and two-dimensional lattices. (C) Cryo-EM density map of the CTLA-4-CD80 lattice, with refined atomic models of the CTLA-4-CD80 complex fitted into the map. CTLA-4 is shown in red, whereas CD80 is shown in purple. (D) Close-up view of the CTLA-4-CD80 complex within the two-dimensional lattice. The region outlined by the dashed cyan box in (C) is enlarged. The second CTLA-4 and CD80 monomers are indicated by primes. (E) Close-up view of the CTLA-4-CTLA-4 interaction interface within the CTLA-4-CD80 lattice. The region outlined by the dashed green box in (D) is enlarged. Residue numbers for the second CTLA-4 monomer are indicated by prime symbols.

Article Snippet: The gene encoding full-length CTLA-4 with an ALFA affinity tag was synthesized (Twist Bioscience) and cloned into the pEG BacMam transfer vector (Table S1) .

Techniques: Cryo-EM Sample Prep, Residue

Journal: bioRxiv

Article Title: Structural basis of CD28 and CTLA-4 interactions with CD80, CD86, and the CD80-PD-L1 heterodimer on artificial and cellular membranes

doi: 10.64898/2026.03.30.715283

Figure Lengend Snippet: (A) Schematic of the lipid monolayer experiment using cellular membrane fragments. An octa-histidine tag was fused to the C-terminus of the CD80 ectodomain, enabling the tethering of the CTLA-4fl-CD80 complex to the Ni²⁺-NTA lipid monolayer. “H” denotes the octa-histidine tag, and the Ni²⁺-NTA lipids are shown in green. Other proteins, carbohydrates, and cholesterols on the cellular membrane fragments are schematically shown as cyan, blue, and orange lines, respectively. (B) Representative cryo-EM micrograph of the CTLA-4fl-CD80 lattice. The regions outlined by magenta lines are enlarged in the lower panels. (C) 2D class averages of linear CTLA-4fl-CD80 clusters and two-dimensional lattices. (D) Cryo-EM density map of the CTLA-4fl-CD80 lattice. (E) Structure of the CTLA-4fl-CD80 lattice on the cellular membrane fragment. The atomic model of the CTLA-4-CD80 complex is fitted into the cryo-EM density map. CTLA-4 is shown in red, whereas CD80 is shown in purple.

Article Snippet: The gene encoding full-length CTLA-4 with an ALFA affinity tag was synthesized (Twist Bioscience) and cloned into the pEG BacMam transfer vector (Table S1) .

Techniques: Membrane, Cryo-EM Sample Prep

Journal: bioRxiv

Article Title: Structural basis of CD28 and CTLA-4 interactions with CD80, CD86, and the CD80-PD-L1 heterodimer on artificial and cellular membranes

doi: 10.64898/2026.03.30.715283

Figure Lengend Snippet: (A) Schematic of the lipid monolayer experiment. An octa-histidine tag was fused to the C-termini of the CD80 and PD-L1 ectodomains. Purified CD80 and PD-L1 were mixed and tethered to a Ni²⁺-NTA lipid monolayer, followed by the addition of CTLA-4 to form the final CTLA-4-CD80-PD-L1 complex. “H” denotes the octa-histidine tag, and the Ni²⁺-NTA lipids are shown in green. (B) Representative cryo-EM micrograph and 2D class averages of the CTLA-4-CD80-PD-L1 complex assembled at a 1:1:1 molar ratio. (C) Representative cryo-EM micrograph and 2D class averages of the CTLA-4-CD80-PD-L1 complex assembled at a 1:1:3 molar ratio. Schematic representations of the CTLA-4-CD80-PD-L1 complexes corresponding to each 2D class are shown below the class averages. (D) Cryo-EM density map of the CTLA-4-CD80-PD-L1 complex. CTLA-4, CD80, and PD-L1 are shown in red, purple and light green, respectively. The planar lipid layer is drawn as a grey parallelogram. (E) Overall structure of the CTLA-4-CD80-PD-L1 complex. The IgV and IgC domains of PD-L1 are labeled. (F) Close-up view of the CD80-PD-L1 interaction interface. The region outlined by the dashed red box in (E) is enlarged.

Article Snippet: The gene encoding full-length CTLA-4 with an ALFA affinity tag was synthesized (Twist Bioscience) and cloned into the pEG BacMam transfer vector (Table S1) .

Techniques: Purification, Cryo-EM Sample Prep, Labeling

Journal: bioRxiv

Article Title: Structural basis of CD28 and CTLA-4 interactions with CD80, CD86, and the CD80-PD-L1 heterodimer on artificial and cellular membranes

doi: 10.64898/2026.03.30.715283

Figure Lengend Snippet: CD28 (orange), CTLA-4 (red), CD80 (purple), CD86 (blue), and PD-L1 (green) are illustrated above. Structural data indicate that CD28 forms a discrete complex with CD80, whereas CTLA-4 displaces CD28 from this complex to promote the formation of higher-order linear and two-dimensional CD80 clusters. PD-L1 binding to CD80 disrupts CD80 homodimerization and remodels the CTLA-4–CD80 assembly into a distinct, “open-leg” CTLA-4–CD80–PD-L1 complex. These observations suggest a model in which CTLA-4-mediated clustering facilitates inhibitory receptor organization and CD80 capture, while PD-L1-mediated de-clustering attenuates CTLA-4–CD80 clustering and alters downstream T-cell signaling. Notably, the PD-L1–CD80 heterodimer preserves the ability of CD80 to bind to CD28.

Article Snippet: The gene encoding full-length CTLA-4 with an ALFA affinity tag was synthesized (Twist Bioscience) and cloned into the pEG BacMam transfer vector (Table S1) .

Techniques: Binding Assay

Journal: Cell Reports Medicine

Article Title: Neoadjuvant Fc-enhanced anti-CTLA-4 targets Tregs to augment androgen deprivation in high-risk prostate cancer: A randomized phase I trial

doi: 10.1016/j.xcrm.2026.102638

Figure Lengend Snippet: Effect of ADT + Fc-enhanced anti-CTLA-4 on DCs (A) Violin plot of myeloid cluster frequencies as percentage of myeloid cells stratified by patient’s 2-year PSA recurrence status. No recurrence, n = 7; recurrence, n = 4. Two-tailed Student’s t test was utilized to test statistical significance. For this and all violin plots to follow, solid lines denote group medians, while dashed lines denote quartiles. (B) Kaplan-Meier curve representing time-to-PSA recurrence in patients stratified by dendritic cell frequency; 95% confidence intervals shown in the shaded areas. There was one recurrence event in DC-high patients and three recurrence events in DC-low patients. Log rank test was performed to evaluate statistical significance. (C) Schematic of pre-clinical validation experiment in the MycCaP model. (D) Percent change in tumor volume relative to baseline volume prior to ADT stratified by treatment group. Average values are represented by solid lines and standard deviations are represented by shaded areas. (E) Kaplan-Meier curve representing survival of mice shown in (D). Log rank (Mantel-Cox) test was used to evaluate statistical significance. (F) Left: supervised UMAP of live CD45 + cells from MycCaP tumors harvested following therapy as denoted in (D and E). Semi-supervised clustering was performed using FlowSOM and clusters were manually annotated according to expression of lineage-defining markers. Right: heatmap of lineage-defining markers across all clusters. Data represent geometric MFI for each marker normalized to the minimum and maximum values across all clusters. (G and H) (G) Frequency of TI-Tregs and (H) frequency of cDCs as percentage of all live CD45 + cells, stratified by treatment group. (I) Left: PaCMAP representing FlowSOM-derived cDC clusters. Clusters were manually annotated according to expression of canonical marker proteins. Right: pseudocolor representation of cDC phenotypes in PaCMAP space stratified by treatment group. (J) Violin plot representing frequency of PD-L2 + DCs as percentage of all DCs stratified by treatment group. (K) Violin plot representing frequency of CD40 + DCs as percentage of all DCs stratified by treatment group. Welch’s t test was used to assess statistical significance. Murine data shown are n = 8 mice per group and representative of two independent experiments each for survival and immune profiling studies.

Article Snippet: Anti-Human CTLA-4 (14D3) 170Er , Standard BioTools , Cat# 3170005B.

Techniques: Two Tailed Test, Biomarker Discovery, Expressing, Marker, Derivative Assay