Listeriolysin is processed efficiently into an MHC class I-associated epitope in Listeria monocytogenes-infected cells.
Listeria monocytogenes is an intracellular pathogen that enters the cytoplasm of infected cells by secreting listeriolysin (LLO), a protein that destroys the phagosomal membrane. In infected mice, LLO is a major Ag detected by protective, MHC class I-restricted CTLs. Although the role of LLO in pathogenesis and host immunity is well established, its rate of intracellular synthesis has yet to be determined. Herein we show that cytosolic L. monocytogenes secrete LLO at a relatively low rate of approximately one molecule per bacterium per minute. Under extracellular labeling conditions, the rate of LLO secretion is approximately 50-fold higher. Intracellular LLO synthesis suffices, however, for the accumulation of 600 to 1000 H-2Kd-associated LLO 91-99 epitopes per cell. We calculate that between four and 11 LLO molecules are degraded for each LLO 91-99 epitope bound by H-2Kd. Our findings indicate that the antigenicity of LLO, with respect to MHC class I-restricted CTLs, cannot be attributed to high levels of intracellular secretion. Rather, LLO is a dominant Ag because it is rapidly degraded and very efficiently processed into an MHC class I-associated epitope.
Villanueva
M S
MS
Section of Infectious Diseases, Yale University School of Medicine, New Haven, CT 06520, USA.
Sijts
A J
AJ
Pamer
E G
EG
D000818:Animals; D017951:Antigen Presentation; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000276:immunology / Q000378:metabolism; D001427:Bacterial Toxins; D002460:Cell Line; D015180:Electrophoresis, Gel, Two-Dimensional; D000939:Epitopes / Q000378:metabolism*; D006183:H-2 Antigens / Q000276:immunology*; D006360:Heat-Shock Proteins / Q000096:biosynthesis* / Q000276:immunology / Q000378:metabolism; D006460:Hemolysin Proteins; D008264:Macrophages / Q000378:metabolism; D008407:Mast Cells / Q000378:metabolism; D051379:Mice; D011233:Precipitin Tests; D014407:Tumor Cells, Cultured
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D000939:Epitopes; D006183:H-2 Antigens; D006360:Heat-Shock Proteins; D006460:Hemolysin Proteins; C038705:hlyA protein, Listeria monocytogenes