Figure 2. Parabiosis surgery reveals... | Rockefeller University Press

Figure 2.

$Parabiosis surgery reveals that a substantive fraction of influenza-specific memory CD8+ T cells in the medLN are TRM. (A) Left: C57/Bl6 (CD45.2) recipient mice received CD45.1+ P14 or Thy1.1+ P14 cells 1 d before i.n. infection with PR8-gp33. 33 d later, mice were conjoined by parabiosis for an additional 23 d before sacrifice (SAC) to allow equilibration of the circulating memory compartment between parabiotic partners. Right: Phenotype of cells to be analyzed in subsequent panels: Q1 (CD103− Ly6Chi), Q2 (CD103+ Ly6Chi), Q3 (CD103+ Ly6Clo), and Q4 (CD103− Ly6Clo). (B) I.v.neg P14 cells isolated from lungs. (C) Representative flow cytometry plots demonstrating the degree of equilibration among cells in Q1 (CD103− Ly6Chi) and Q3 (CD103+ Ly6Clo) within host medLNs. H, host-derived; P, partner-derived P14 cells. (D) Enumeration of host and partner P14 cells (left) and percentage resident (right) within the indicated tissues or phenotypic subsets present in the medLN of parabionts. Data are representative of 12 parabiotic partners (6 pairs) analyzed over two independent experiments. *, P < 0.05; ****, P < 0.0001 as determined by one-way ANOVA comparing CD103− Ly6Chi P14 cells in Q1 (denoted by symbol ¤) to any other tissue. Bars represent mean ± SEM. (E) Ratio of host- and partner-derived P14 cells isolated from peripheral blood, right ventricular blood, or the i.v.pos fraction of the lung (n = 4).$

Parabiosis surgery reveals that a substantive fraction of influenza-specific memory CD8⁺ T cells in the medLN are T_RM. (A) Left: C57/Bl6 (CD45.2) recipient mice received CD45.1⁺ P14 or Thy1.1⁺ P14 cells 1 d before i.n. infection with PR8-gp33. 33 d later, mice were conjoined by parabiosis for an additional 23 d before sacrifice (SAC) to allow equilibration of the circulating memory compartment between parabiotic partners. Right: Phenotype of cells to be analyzed in subsequent panels: Q1 (CD103⁻ Ly6C^hi), Q2 (CD103⁺ Ly6C^hi), Q3 (CD103⁺ Ly6C^lo), and Q4 (CD103⁻ Ly6C^lo). (B) I.v.^neg P14 cells isolated from lungs. (C) Representative flow cytometry plots demonstrating the degree of equilibration among cells in Q1 (CD103⁻ Ly6C^hi) and Q3 (CD103⁺ Ly6C^lo) within host medLNs. H, host-derived; P, partner-derived P14 cells. (D) Enumeration of host and partner P14 cells (left) and percentage resident (right) within the indicated tissues or phenotypic subsets present in the medLN of parabionts. Data are representative of 12 parabiotic partners (6 pairs) analyzed over two independent experiments. *, P < 0.05; ****, P < 0.0001 as determined by one-way ANOVA comparing CD103⁻ Ly6C^hi P14 cells in Q1 (denoted by symbol ¤) to any other tissue. Bars represent mean ± SEM. (E) Ratio of host- and partner-derived P14 cells isolated from peripheral blood, right ventricular blood, or the i.v.^pos fraction of the lung (n = 4).