The Trypanosoma cruzi membrane glycoprotein AGC10 inhibits human lymphocyte activation by a mechanism preceding translation of both, interleukin-2 and its high-affinity receptor subunits.
Like living Trypanosoma cruzi, its AGC10 membrane glycoprotein inhibits interleukin-2 (IL-2) secretion and membrane expression of CD25, CD122, and CD132 (the components of the high-affinity IL-2 receptor) by activated human lymphocytes. Since these molecules are required for effective lymphocyte division, we explored the molecular mechanism underlying these alterations. In the presence of AGC10 the cytoplasmic levels of IL-2 protein of CD4(+) and CD8(+) blood lymphocytes stimulated with phorbol myristate acetate (PMA) plus ionomycin were markedly reduced. AGC10 also decreased the intracellular levels of CD25, CD122, and CD132 in CD4(+) and CD8(+) cells stimulated with the T-cell mitogen phytohemagglutinin (PHA). These results indicated that the AGC10-induced alterations preceded IL-2 secretion and transport of IL-2 receptor components to the cell membrane. Supporting this view were the substantially diminished levels of IL-2, CD25, CD122, and CD132 mRNA found in AGC10-containing cultures of PHA-activated lymphocytes. These decreases were not due to increased mRNA instability. Thus, the rates of decay for each of these mRNA species were comparable in the presence or absence of AGC10, suggesting a mechanism involving transcription inhibition. AGC10 targeted an early lymphocyte activation event since inhibition of lymphoproliferation subsided when AGC10 was added to cultures at or after 20 h post-activation. AGC10 also caused large reductions in the mRNA levels of cyclin D2 and cdk4, both critical for progression through G1. These results show for the first time that AGC10-induced inhibition of lymphoproliferation entails curtailed biosynthesis of IL-2 and, IL-2 receptor molecules, and suggest that the effect involves inhibition of gene transcription.
Kierszenbaum
Felipe
F
Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, MI 48824, USA. kierszen@msu.edu
Fresno
Manuel
M
Sztein
Marcelo B
MB
D000818:Animals; D002453:Cell Cycle; D002478:Cells, Cultured; D056742:Cyclin D2; D016213:Cyclins / Q000032:analysis; D016207:Cytokines / Q000096:biosynthesis / Q000145:classification; D016193:G1 Phase / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects / Q000276:immunology; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D008213:Lymphocyte Activation / Q000187:drug effects*; D008562:Membrane Glycoproteins / Q000378:metabolism / Q000494:pharmacology*; D014176:Protein Biosynthesis; D015800:Protozoan Proteins / Q000378:metabolism / Q000494:pharmacology*; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D015375:Receptors, Interleukin-2 / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D014349:Trypanosoma cruzi / Q000737:chemistry* / Q000166:cytology
C080336:CCND2 protein, human; D056742:Cyclin D2; D016213:Cyclins; D016207:Cytokines; D007376:Interleukin-2; D008562:Membrane Glycoproteins; D015800:Protozoan Proteins; D012333:RNA, Messenger; D015375:Receptors, Interleukin-2