Use of colony assays and anti-T cell immunotoxins to elucidate the immunobiologic features of leukemic progenitor cells in T-lineage acute lymphoblastic leukemia.
The specific binding of radioiodinated rIL-2 to fresh marrow blasts from T-lineage acute lymphoblastic leukemia (ALL) patients was initially investigated. The estimated number of radioiodinated rIL-2 molecules bound per blast ranged from undetectable to 1948. In colony assays, 72% of 32 cases analyzed showed a significant proliferative response to rIL-2, which depended on PHA-stimulated lymphocyte conditioned medium activation. Colony stimulation indices correlated with the number of radioiodinated rIL-2 molecules bound per blast but not with expression of CD25/Tac Ag on fresh marrow blasts or primary colony blasts. These findings provide evidence that in T-lineage ALL functional IL-2R proteins are expressed on leukemic progenitor blasts which may be distinct from Tac Ag. We used the mAb 35.1, T101, and G3.7 to test for expression of CD2, CD5, and CD7 on fresh marrow blasts from 126 T-lineage ALL patients. CD2, CD5, and CD7 were expressed in 84%, 93%, and 99% of cases, respectively. Furthermore, colony blasts that represent the early progeny of leukemic progenitor blasts were also CD2+CD5+CD7+. Ricin conjugates of 35.1, T101, and G3.7 mAb were used as Ag-specific cytotoxic probes to test for expression of CD2, CD5, and CD7 at the level of T-lineage leukemic progenitor blasts. Each immunotoxin was able to selectively eliminate greater than 99% of leukemic progenitor blasts, providing unique and direct evidence that these cells co-express CD2, CD5, and CD7. Neither mixtures of anti-CD5 and anti-CD7 nor anti-CD2, anti-CD5, and anti-CD7 immunotoxins were more effective against blast progenitor cells than the individual immunotoxins alone, confirming that CD2, CD5, and CD7 are not expressed on non-overlapping progenitor cell subpopulations.
D000911:Antibodies, Monoclonal / Q000276:immunology; D000945:Antigens, Differentiation, T-Lymphocyte / Q000032:analysis; D003114:Colony-Forming Units Assay; D006412:Hematopoietic Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D000922:Immunotoxins / Q000494:pharmacology*; D007376:Interleukin-2 / Q000378:metabolism; D007945:Leukemia, Lymphoid / Q000473:pathology*; D008213:Lymphocyte Activation; D014411:Neoplastic Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D010641:Phenotype; D011971:Receptors, Immunologic / Q000096:biosynthesis*; D015375:Receptors, Interleukin-2; D011994:Recombinant Proteins / Q000378:metabolism; D012276:Ricin / Q000494:pharmacology*; D013601:T-Lymphocytes; D014410:Tumor Stem Cell Assay
D000911:Antibodies, Monoclonal; D000945:Antigens, Differentiation, T-Lymphocyte; D000922:Immunotoxins; D007376:Interleukin-2; D011971:Receptors, Immunologic; D015375:Receptors, Interleukin-2; D011994:Recombinant Proteins; D012276:Ricin