Enhancement by monocytes of perforin production and its gene expression by human CD8+ T cells stimulated with interleukin-2.
Pore-forming protein (PFP) is an important effector molecule for cytotoxicity mediated by cytotoxic T cells and NK cells. In the present study, the effect of monocytes on PFP production by interleukin-2 (IL-2)-stimulated T lymphocytes was examined. Highly purified lymphocytes (> 99%) and monocytes (> 90%) were isolated by centrifugal elutriation from peripheral blood of healthy donors, and, CD4+ and CD8+ cells were isolated from the purified lymphocytes by using antibody-bound magnetic beads. PFP production was quantitated with a universal microspectrophotometer in combination with immunostaining using anti-PFP antibody. Monocytes did not produce any PFP. High levels of PFP production were observed in CD8+ cells, but not CD4+ cells after incubation for 4 days with IL-2. Addition of monocytes to cultures of CD8+ cells resulted in significant augmentation of PFP production after 3 days' stimulation with IL-2. Monokines (TNF alpha and IL-6) caused a significant increase in PFP production by IL-2-stimulated CD8+ cells. Northern blot analysis revealed that the PFP mRNA levels was enhanced by stimulation with IL-2, and that addition of monocytes to cultures of CD8+ cells plus IL-2 augmented their PFP mRNA expression. These observations suggest that monocytes are important in in situ regulation of the CD8+ T cell-mediated cytotoxic response through production of PFP.
Third Department of Internal Medicine, University of Tokushima School of Medicine.
K
K
Sugihara
S
S
Sone
M
M
Shono
A
A
Nii
M
M
Munekata
K
K
Okumura
T
T
Ogura
D000328:Adult; D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects / Q000378:metabolism; D016827:CD8 Antigens / Q000502:physiology*; D015870:Gene Expression / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D007376:Interleukin-2 / Q000494:pharmacology*; D007700:Kinetics; D008213:Lymphocyte Activation / Q000187:drug effects / Q000502:physiology; D008297:Male; D008562:Membrane Glycoproteins / Q000096:biosynthesis* / Q000235:genetics*; D009000:Monocytes / Q000502:physiology*; D054353:Perforin; D052899:Pore Forming Cytotoxic Proteins; D012333:RNA, Messenger / Q000235:genetics; D013268:Stimulation, Chemical; D013601:T-Lymphocytes / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D015854:Up-Regulation / Q000187:drug effects / Q000502:physiology
D016827:CD8 Antigens; D007376:Interleukin-2; D008562:Membrane Glycoproteins; D052899:Pore Forming Cytotoxic Proteins; D012333:RNA, Messenger; D054353:Perforin