3A4, a new potential target for B and myeloid lineage leukemias.

摘要

Antibody-targeting therapy has drawn great interests to the hematologists and oncologists. Many antibodies have been studied for their potential targeting for hematopoietic malignancies. A few have been proved to be very effective for patients with these diseases. However, more antibodies are needed for clinical use. CD45 and its isoforms may convey clinical potential in terms of targeting therapy. Zhejiang Children's Hospital (ZCH)-6-3A4 (3A4), a novel antibody that can recognize an isoform of CD45 has been found to react with restricted cell components in hematopoietic system, which may have the potential for targeting therapy. Herein, we conducted an in vitro study of our newly prepared antibody 3A4 using various cellular and immunocytological methods. The results showed that the antibody 3A4 (murine IgG1kappa) was a new clone of anti-CD45RA. It could block the binding to an epitope of CD45RA recognized by a standard anti-CD45RA antibody (Clone name L48). The reactivity of the 3A4 to both fresh leukemia cells from patients and well-defined leukemia cell lines was largely similar to those of L48, but the former recognized more leukemia cells than the latter. Cytometric analysis after papain treatment showed that the internalization rate of the 3A4 antibody to the target cells was as high as 71.3% after incubation at 37 degrees C for 4 h, which was significantly higher than that of L48 (20.4%). The norcantharidin (NCTD)-conjugated immunotoxin (NCTD-3A4) was generated using an active ester method. The targeting inhibition rate on KG1a was as high as 61.10% after 96 h incubation in a dose-dependent manner, which was significantly higher than that (3.56%, P < 0.01) with 3A4-negative Nalm-6 cells. In conclusion, our new anti-CD45RA antibody 3A4 is probably a new target molecule of leukemia cells and holds a targeting therapeutic potential for hematopoietic malignancies, which warrants further development of this agent.