Expt | Condition | T cell: APC ratio giving 50% maximum proliferation of OT-I T cellsab
|
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L-DC | Resi Mono | Infl mono | Neu | cDC |
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I | + LPS | 33 | - | - | 33 | 300 |
- LPS | 33 | - | - | <33 | 300 |
II | + LPS | 33 | 33 | - | - | 300 |
- LPS | 33 | <33 | - | - | 300 |
III | + LPS | 100 | 42 | - | - | 300 |
- LPS | 100 | 42 | - | - | 141 |
IV | + LPS | 100 | 42 | <33 | - | 300 |
- LPS | 100 | 33 | <33 | - | 300 |
V | - LPS | 100 | 33 | 0 | <33 | - |
VI | + LPS | 141 | 0 | - | <33 | - |
- LPS | 100 | 0 | - | <33 | - |
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aFor preparation of APC, splenocytes were harvested from Act-mOVA mice and prepared by red blood cell lysis followed by T and B cell depletion. L-DC, cDC, resident monocytes (Resi mono), inflammatory monocytes (Infl mono), neutrophils (Neu) were sorted as described in Table 1
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b APC, with and without LPS (10ug/ml), were cocultured with 105 CFSE-labelled OT-I (TCR-tg) CD8+ T cells, sorted as PI−Thy1.2+Vα2+CD4− cells, at T cell:APC ratios of 33, 100, 300 and 900:1. After 72 h, CD8+ OT-I T cells were gated as PI−CD11b−Thy1.2+Vα2+ cells, and CFSE dilution assessed flow cytometrically to estimate % proliferating T cells. OT-I T cells alone served as a control (Con)