Purification and further characterization of a non-tumor necrosis factor alpha or beta differentiation-inducing cytokine, P48.
In this report, we present the further characterization and purification of a cytokine differentiation factor, termed P48, which unlike previously described differentiation factors is antigenically unrelated to tumor necrosis factor alpha (TNF-alpha), tumor necrosis factor beta (TNF-beta), and gamma interferon. HL-60 cells and phorbol diester-resistant HL-60-1E3 cells exposed to conditioned medium from Reh cells mature along the monocyte/macrophage pathway, as assessed by several assays (express nonspecific esterase, produce superoxide anion, morphologically resemble monocytes, mediate phorbol diester-triggered extracellular cytolytic activity). Reh cell conditioned medium is antiproliferative toward a panel of cell lines, is not nonspecifically cytotoxic, has no antiviral or colony-stimulating factor activities, and is not affected by exposure to insolubilized anti-gamma interferon. A 48-kDa glycoprotein (P48) which mediates this differentiation factor activity has been purified to homogeneity from Reh cell conditioned medium, and a polyclonal neutralizing antiserum has been produced. P48 activity is not blocked by either anti-TNF-alpha and anti-TNF-beta and on Western blot analysis is antigenically distinct from TNF-alpha and TNF-beta. In addition, polyclonal anti-P48 does not block either TNF-alpha or TNF-beta activities or recognize either on Western blots. Unlike gamma interferon, colony-stimulating factor, TNF-alpha, or TNF-beta, P48 reverses phorbol diester resistance of HL-60-1E3 cells. These studies present strong evidence for the existence of a previously unrecognized cytokine which, unlike other reported differentiation factors, is antigenically unrelated to TNF-alpha or TNF-beta. P48 may play an important role in growth and development of normal and abnormal (leukemic) hematopoietic and nonhematopoietic cells.
Leftwich
J A
JA
Laboratory of Tumor Immunology, Guthrie Clinic/Medical Center, Sayre, Pennsylvania 18840-1692.
Hall
R E
RE
D000818:Animals; D000906:Antibodies / Q000276:immunology; D000918:Antibody Specificity; D001685:Biological Factors / Q000276:immunology / Q000302:isolation & purification* / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects*; D003470:Culture Media / Q000494:pharmacology; D016207:Cytokines; D004305:Dose-Response Relationship, Drug; D005622:Friend murine leukemia virus; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute / Q000473:pathology; D007942:Leukemia, Experimental / Q000473:pathology; D051379:Mice; D015240:Phorbol 12,13-Dibutyrate; D011506:Proteins / Q000276:immunology / Q000302:isolation & purification* / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors / Q000276:immunology
D000906:Antibodies; D001685:Biological Factors; D003470:Culture Media; D016207:Cytokines; D011506:Proteins; D014409:Tumor Necrosis Factor-alpha; D015240:Phorbol 12,13-Dibutyrate