Figure 6. Expansion of Nb-T2 T-cell receptor (TCR) cells in mesenteric lymph node (MLN) after Nippostrongylus brasiliensis (Nb) infection.

Hematopoietic stem cells were transduced with retroviral vectors for expression of potential Nb-specific clones (Nb-T1, Nb-T2, and Nb-T3) and used to generate retrogenic mice. (A) UMAP overlay shows distribution of TCRs selected for retrogenic expression. V gene segments, CDR3 sequence, and distribution between organs for selected lung-expanded TCR clones (Nb-T1, Nb-T2, and Nb-T3). (B) Eight weeks after reconstitution of irradiated Rag1KO mice with TCR chain and fluorescent marker-encoding retrovirus containing retrogenic Rag1 stem cells, T cells were harvested, transferred to Ly5.1 B6 wild-type mice which were then infected with Nb or Heligmosomoides polygyrus (Hp) as control. (C) Plots show the percentage of TCR transgenic cells (GFP⁺, Thy1.1⁺) among live CD4⁺ cells from constructs Nb-T1, Nb-T2, and Nb-T3 in the MLN of recipient mice at day 9 post infection. (D) Percentage and total number of Nb-T2 cells in MLN. Quantification is based on six independent experiments. Statistical significance determined by Mann–Whitney U test; **p < 0.001.

Figure 6—figure supplement 1. Differentiation into CD4⁺ T-cell receptor (TCR) transgenic cells.

(A) Composition of TCRα and TCRβ chains of TCRs to be overexpressed in TCR retrogenic mice. (B) Eight weeks after reconstitution of irradiated Rag1KO mice, T-cell development was evaluated by flow cytometry in spleen and mesenteric lymph node (MLN). TCR expression is inferred by expression of GFP (TCRα chain) and Thy1.1 (TCRβ chain). Pre-gating on living singlets was performed. Further gates are applied to distinguish between CD4 and CD8 expressing cells. Representative plots are shown.