CloneID: 7D4
Antigen Long Description: The original rat IgM version of this antibody was raised by immunizing Lewis rats with HT2 cells in complete Freund's adjuvant via a intraperitoneal injection and subsequent generation of hybridomas.
Buffer Composition: PBS only.
Chimeric Use Statement: This chimeric mouse antibody was made using the variable domain sequences of the original Rat IgM format, for improved compatibility with existing reagents, assays and techniques.
Uniprot Accession No.: P01590
Specificity Statement: 7D4 recognizes mouse CD25 at an epitope comprising amino acid 184 to 194 (REHHRFLASEE). This epitope is distinct from the IL-2 binding site and does not block binding of IL-2 to CD25. Murine CD25 is a 55 kDa interleukin-2 receptor alpha chain (IL-2R alpha). CD25 is expressed by early progenitors of the T and B lineage as well as by activated mature T and B lymphocytes. By itself, CD25 binds IL-2 only with low affinity. However, CD25 associates with CD122 (IL-2 receptor beta chain) and CD132 (common gamma chain) to form the high affinity IL-2 receptor.
Application Notes (Clone): 7D4 is a non-IL-2 blocking anti-CD25 antibody that has been widely used to identify CD25-positive cells via flow cytometry (Kohyama et al., 2012; pmid: 22399592), for instance, to analyse Lymph node cells from BALB/c mice that were untreated or treated with 0.25 mg PC61 (Onizuka et al., 1999; pmid: 10397255) or to characterize brain-infiltrating immune cells (Maes et al., 2013). Although 7D4 does not inhibit IL-2 binding itself, it shows synergistic qualities with a cancer vaccine; namely, 7D4 with a cancer vaccine boosted the vaccine-induced anti-tumour response. Furthermore, this antibody was shown to be able to induce inactivation of regulatory T cells (Treg); the effect was particularly significant when this clone was used synergistically with another anti-CD25 clone - PC61 (Ab01107) (Kohm et al., 2006; pmid: 16517695; McNeill et al., 2007; pmid: 17212768). 7D4, when turned into a mouse IgG2a format, exhibits depleting qualities against Treg cells and can be used to remove Treg lymphocytes from the tumoral environment without blocking CD25 (IL-2 pathway) on T effector cells (Solomon et al., 2020). As a result, it can be utilized as a potential immunotherapy agent achieving Treg depletion without T effector cells' debilitation (Solomon et al., 2020). Finally, it was demonstrated that such anti-CD25 non-IL-2 blocking antibodies are successful agents in promoting effector activation and leading to strong antitumor activity (Solomon et al., 2020).
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