Anti-Human CD2 Recombinant Antibody (BTI-322 (also known as Lo-CD2a)) (CAT#: TAB-1631CL)

Figure 1 Effect of BTI-322 on primary and secondary xenogeneic MLR.

(a) Human PBMCs (1 × 10⁶ /ml) were cultured at a 1 : 1 ratio with irradiated SLAᵈᵈ porcine PBMCs, in the presence of 200 ng/ml BTI-322 or the humanized version MEDI-507, or control isotype-matched Ig, followed by measurement of [³H]-TdR incorporation (cpm) at days 3, 5, and 7. (b) Cells harvested after primary xenogeneic stimulation in the presence of these antibodies or controls were cultured for 3 days without stimulant, and then subjected to a secondary xenogeneic MLR, followed by measurement of [³H]-TdR incorporation (cpm) at days 3 and 5.

Xu, Y., Kolber‐Simonds, D., Hope, J. A., Bazin, H., Latinne, D., Monroy, R.,... & SCHUURMAN, H. J. (2004). The anti‐CD2 monoclonal antibody BTI‐322 generates unresponsiveness by activation‐associated T cell depletion. Clinical & Experimental Immunology, 138(3), 476-483.

Figure 2 Use of T cells as responder cells and effect of monocytes.

(a)PBMCs or nylon-wool-purified T cells (both 1 × 10⁶ /ml) were cultured at a 1 : 1 ratio with irradiated SLAᵈᵈ porcine PBMCs, in the presence of 200 ng/ml BTI-322 or control Ig, followed by measurement of [³H]-TdR incorporation (cpm) at days 3, 5, and 7. (b) Purified responder T cells were supplemented with monocytes (adherent cells) at relative proportions [100% reflects the original proportion in the original PBMC preparation (10-20%)].

Xu, Y., Kolber‐Simonds, D., Hope, J. A., Bazin, H., Latinne, D., Monroy, R.,... & SCHUURMAN, H. J. (2004). The anti‐CD2 monoclonal antibody BTI‐322 generates unresponsiveness by activation‐associated T cell depletion. Clinical & Experimental Immunology, 138(3), 476-483.

Figure 3 Effect of BTI-322 on secondary MLR: SLAᶜᶜ, third-party SLAᵈᵈ and allogeneic restimulation after primary stimulation with SLAᵈᵈ cells.

(a) Cells (1 × 10⁶/ml) from primary 7-day xenogeneic MLR (1 : 1 responder : stimulator ratio) in the presence of 200 ng/ml BTI-322 or control Ig were cultured in secondary MLR with cells of the original SLA haplotype, third-party xenogeneic cells and allogeneic cells. Data shown are [³H]-TdR incorporation at various time-points during secondary culture,or cells cultured during the primary MLR. (b) The same experiment, using allogeneic stimulation in primary MLR. (c) The same experiment using suboptimal xenogeneic stimulation in primary MLR (1 : 0.125 responder : stimulator ratio).

Xu, Y., Kolber‐Simonds, D., Hope, J. A., Bazin, H., Latinne, D., Monroy, R.,... & SCHUURMAN, H. J. (2004). The anti‐CD2 monoclonal antibody BTI‐322 generates unresponsiveness by activation‐associated T cell depletion. Clinical & Experimental Immunology, 138(3), 476-483.

Figure 4 Effect of BTI-322 on primary and secondary stimulation with anti TCR Vβ family-specific antibodies.

(a) PBMCs were incubated in primary culture with 10 μg/ml anti-TCR Vβ8 antibody, with or without BTI-322 or control Ig (100 ng/ml): [³H]-TdR incorporation was measured at day 7. (b) Cells after primary stimulation as in (a) were washed and cultured for 3 days without stimulator, and then subjected to secondary stimulation with the original anti-Vβ8 antibody or an irrelevant anti-Vβ13 antibody (10 μg/ml): [³H]-TdR incorporation was measured at day 3.

Xu, Y., Kolber‐Simonds, D., Hope, J. A., Bazin, H., Latinne, D., Monroy, R.,... & SCHUURMAN, H. J. (2004). The anti‐CD2 monoclonal antibody BTI‐322 generates unresponsiveness by activation‐associated T cell depletion. Clinical & Experimental Immunology, 138(3), 476-483.

Figure 5 Presence of CD3⁺Vβ8⁺ cells after stimulation of PBMCs with anti-TCR Vβ8 or anti-TCR Vβ13 antibody in the presence of BTI- 322 or control Ig.

PBMCs (1 × 10⁶/ml) were stimulated for 7 days by control antibody (a,b), anti-Vβ8 antibody (c,d) or anti-Vβ13 antibody (e,f) at 100 ng/ml, in the presence of 100 ng/ml control antibody (a,c,e) or BTI-322 (b,d,f). Subsequently, flow cytometry was performed with a fluorescein isothiocyanate (FITC)-conjugated anti-CD3 antibody in combination with an anti-TCR Vβ8 antibody [indirect staining using a phycoerythrin (PE)-conjugated secondary antibody]. The position of the CD3⁺Vβ8⁺ double-positive population in the individual plots is indicated by circles, determined by flow cytometry on freshly isolated PBMCs from the same donor with appropriate isotypematched control antibody and single-antibody staining. The CD3⁺Vβ8⁺ double-positive population comprises approximately 4.5% of the viable cells in (a,b,e and f); approximately 14% in (c); and approximately 1% in (d).

Xu, Y., Kolber‐Simonds, D., Hope, J. A., Bazin, H., Latinne, D., Monroy, R.,... & SCHUURMAN, H. J. (2004). The anti‐CD2 monoclonal antibody BTI‐322 generates unresponsiveness by activation‐associated T cell depletion. Clinical & Experimental Immunology, 138(3), 476-483.