Rat Anti-Cd8b1 Recombinant Antibody (clone YTS156.7.7) (CAT#: HPAB-S0182-YC)

Figure 1 CD8-β is a target of ART2.2-mediated ADP-ribosylation.

In the liver, the CD8-α⁺ T cells displayed strong staining with the anti-CD8-β mAb H35-17.2, but reduced staining with the anti-CD8-β mAb YTS156.7.7 and 53-5.8.

Lischke, T., Schumacher, V., Wesolowski, J., Hurwitz, R., Haag, F., Koch‐Nolte, F., & Mittrücker, H. W. (2013). CD 8‐β ADP‐ribosylation affects CD 8+ T‐cell function. European Journal of Immunology, 43(7), 1828-1838.

Figure 2 The anti-CD8-β mAb clones YTS156.7.7 and 53-5.8 do not recognize CD8-β in the presence of extracellular NAD⁺.

ART2.2 transfers ADP-ribose from extracellular NAD⁺ onto arginine residues of adjacent surface proteins. Incubation of splenic CD8+ T cells with NAD⁺ resulted in a concentrationdependent loss of binding of the anti-CD8-β mAb YTS156.7.7 and 53-5.8, but not of mAb H35-17.2 or of the anti-CD8-α mAb 53-6.7.

Lischke, T., Schumacher, V., Wesolowski, J., Hurwitz, R., Haag, F., Koch‐Nolte, F., & Mittrücker, H. W. (2013). CD 8‐β ADP‐ribosylation affects CD 8+ T‐cell function. European Journal of Immunology, 43(7), 1828-1838.

Figure 3 CD8-β ADP-ribosylation is stable in vivo for up to 24 h.

Spleen cells were incubated in vitro with 100 μM NAD⁺, and the ADP-ribosylation was determined with anti-CD8-β mAb at different time points after the start of NAD⁺ treatment. After 30 min, we observed substantial loss of staining with the sensitive anti-CD8-β mAb YTS156.7.7 and mAb 53-5.8.

Lischke, T., Schumacher, V., Wesolowski, J., Hurwitz, R., Haag, F., Koch‐Nolte, F., & Mittrücker, H. W. (2013). CD 8‐β ADP‐ribosylation affects CD 8+ T‐cell function. European Journal of Immunology, 43(7), 1828-1838.

Figure 4 ADP-ribosylation of CD8-β is enhanced in the absence of CD38.

Spleen cells from CD38 KO mice showed profoundly reduced staining levels for the sensitive anti-CD8-β mAb YTS156.7.7 and 53-5.8, and treatment of mice with NAD⁺ prior to spleen cell isolation resulted in a further decline in the staining intensity.

Lischke, T., Schumacher, V., Wesolowski, J., Hurwitz, R., Haag, F., Koch‐Nolte, F., & Mittrücker, H. W. (2013). CD 8‐β ADP‐ribosylation affects CD 8+ T‐cell function. European Journal of Immunology, 43(7), 1828-1838.

Figure 5 CD8-β ADP-ribosylation occurs during cell isolation.

For all analyzed tissues, we consistently observed lower staining levels for the sensitive anti-CD8-β mAb for cells from WT mice when compared to cells from ART2 KO mice.

Lischke, T., Schumacher, V., Wesolowski, J., Hurwitz, R., Haag, F., Koch‐Nolte, F., & Mittrücker, H. W. (2013). CD 8‐β ADP‐ribosylation affects CD 8+ T‐cell function. European Journal of Immunology, 43(7), 1828-1838.

Figure 6 YTS156.7.7, but not YTS105.18, blocks MHC class I and CD8 interaction and subsequent T cell activation.

Incubation of OT-1 T cells with YTS156.7 mAb prior to tetramers of H-2K^b/OVA completely blocked calcium flux, indicating inhibition of pMHCI/CD8 interaction.

Shore, D. A., Issafras, H., Landais, E., Teyton, L., & Wilson, I. A. (2008). The crystal structure of CD8 in complex with YTS156. 7.7 Fab and interaction with other CD8 antibodies define the binding mode of CD8 αβ to MHC class I. Journal of molecular biology,384(5), 1190-1202.