Intranasal LCMV antigen was not sufficient to induce medLN TRM after i.p. LCMV infection. (A) Strategy for determining if local antigen presentation in the medLN could drive the formation of medLN TRM de novo. Mice were seeded with P14 and OT-I cells and contemporaneously infected with LCMV i.p. and PR8-ova i.n. ± 3 µg gp33 peptide within the i.n. PR8-ova inoculum. SAC, sacrifice; Spec., specific. (B) Cotransferred transgenic T cells (both P14 and OT-I are Vα2+). (C) Ratio of P14 to OT-I cells ± i.n. gp33 peptide (log2 scale). Solid line indicates 1:1 ratio. While P14 and OT-I cells were similarly expanded in circulation, LCMV-specific cells were underrepresented in the medLN and i.v.neg respiratory tissues. (D) Phenotyping of P14 and OT-I cells recovered from the spleen or medLN of contemporaneously challenged mice given i.n. gp33 peptide at time of infection as in C. n = 3 without gp33; n = 4 with gp33. Bars represent mean ± SEM.
