Journal: bioRxiv

Article Title: Leishmania major co-opts IL-7 feedback in monocytes to suppress CD4⁺ T-cell immunity

doi: 10.64898/2026.03.24.713728

Figure Lengend Snippet: A. Upper panels: H2KB + or PD-L1 + fraction and H2KB or PD-L1 MFI of among fibroblasts from mice reconstituted with Pdpn-cre × Il7r WT/WT littermate control BM (black symbols) or Pdpn-cre × Il7r fl/fl BM (blue symbols). Data normalized to mice reconstituted with littermate control BM. Lower panels: H2KB + or PD-L1 + fraction and H2KB or PD-L1 MFI of among fibroblasts from non-depleted control (black symbols) or Pdpn -dependent 100% depleted (blue symbols) ear infections. Each symbol represents one individual mouse ear. Data normalized to non-depleted control animals. Horizontal lines denote the mean. ***, p<0.001; **, p<0.01; *, p<0.05 according to unpaired t-test. Data shown correspond to the experiments shown in ,, and , respectively. B. Expression of Il7 within L. major -infected (week 3 p.i.) and non-infected ear tissue. Expression as determined by ΔΔCT between Il7 and Hprt qPCR. CT values are corrected according to the mean difference in RNA extracted (see fig. S4C). Each symbol represents one individual mouse ear. Horizontal lines denote the mean. *, p<0.05 according to unpaired t-test. C. Expression level of Prrx among fibroblasts, LEC, BEC and PDPN - CD31 - non-immune cells within L. major -infected ear tissue at week 3 p.i.. Expression as determined by ΔΔCT between Prrx and Hprt qPCR followed by normalization to PDPN - CD31 - cells. Each symbol represents one individual mouse ear. Horizontal lines denote the mean. ****, p<0.0001 according to one-way ANOVA multiple comparison with Dunnett’s post-test. Data in (A-C) pooled from two independent experiments. D. Proportion of fibroblasts, LEC, and BEC among CD45 - TER119 - 7AAD - cells within the L. major infected ear at week 3 p.i.. Each symbol represents one individual mouse ear. Horizontal lines denote the mean. ****, p<0.0001 according to one-way ANOVA multiple comparison with Dunnett’s post-test. E. Experimental setup to test for Prrx -expressing fibroblasts as a necessary source of IL-7 to promote Lm lo Mo3 mediated suppression of CD4 + T cell responses in the L. major infected ear tissue. F. Expression level of Il7 among fibroblasts, LEC, and BEC within L. major -infected ear tissue from littermate control ( Prrx-Cre × Il7 wt/wt ) (black symbols) and Prrx-cre × Il7 fl/fl (blue symbols). Expression as determined by ΔCT between Il7 and Hprt qPCR. G. Left: the fraction of L. major infected cells among recruited monocytes in the ear tissue at week 3 p.i., littermate control ( Prrx-Cre × Il7 wt/wt ) (filled symbols) and Prrx-cre × Il7 fl/fl (open symbols). Differently shaded symbols represent two independent experiments. Each symbol represents one individual mouse ear. Horizontal lines denote the median. ***, p<0.001 according to two-way ANOVA comparing genotypes. Right: Limiting dilution assay (LDA) of L. major tissue burden in the ear. Data normalized to littermate control animals. H. IFNγ + fraction and MFI of IFNγ signal among effector CD4 + T cells within L. major -infected ear at week 3 p.i. of either littermate control (black symbols) or Prrx-cre × Il7 fl/fl animals (blue symbols). Data normalized to littermate control animals. Each symbol in (G-I) represents one individual mouse ear, data are from two independent experiments. Horizontal lines denote the mean. *, p<0.05 according to unpaired t-test.

Article Snippet: The following TaqMan® Gene Expression Assays (Thermo Fisher Scientific) were used according to the manufacturer’s instructions: Il7 (FAM-MGB probe Mm01295804 m1) and Hprt (FAM-MGB probe Mm00446968 m1).

Techniques: Control, Expressing, Infection, Comparison, Limiting Dilution Assay

Journal: bioRxiv

Article Title: Leishmania major co-opts IL-7 feedback in monocytes to suppress CD4⁺ T-cell immunity

doi: 10.64898/2026.03.24.713728

Figure Lengend Snippet: A . Proportion of IFNγ + produced by T, NK, and dendritic cells at week 3 p.i. Each symbol represents one individual mouse ear. Horizontal lines denote the mean. ****, p<0.0001 according to unpaired t-test. B. Expression of the Il7 cytokine gene in ex vivo cultivated fibroblasts without (black symbols) and with (red symbols) addition of IFNγ + for 24h. Horizontal lines denote the median. **, p<0.01 according to Mann-Whitney test. C. Experimental setup to test the impact of anti-IFNγ treatment in fibroblasts within an ongoing L. major infection. D. Correlation (r 2 , with p value indicated) and linear regression (red line) between L. major infection rate (determined by flow cytometry) and Il7 expression level (qPCR) in fibroblasts within the infected ear (week 3 p.i.), in either isotype control (black symbols, upper graph) or anti-IFNγ treated (orange symbols, lower graph) animals. Each symbol represents one replicate of flow cytometry analysis from six individual infected ears per condition with qPCR of up to 3 pools of 200 fibroblasts sorted from the same infected ear. The vertical dashed line shows the threshold defined for analyzing highly infected tissues. E. Comparison of Il7 expression in fibroblasts sorted from tissues with an infection rate above the threshold (dashed line) shown in (D). F. H2KB + or PD-L1 + fraction and H2KB or PD-L1 MFI among fibroblasts isolated from week 3 p.i. infected ears of animals treated with either isotype control (black symbols) or anti-IFNγ antibody (orange symbols). Each symbol in (E-G) represents one individual mouse ear, horizontal lines denote the mean. ***, p<0.001; **, p<0.01; *, p<0.05 according to unpaired t-test. Data were collected in two independent experiments.

Article Snippet: The following TaqMan® Gene Expression Assays (Thermo Fisher Scientific) were used according to the manufacturer’s instructions: Il7 (FAM-MGB probe Mm01295804 m1) and Hprt (FAM-MGB probe Mm00446968 m1).

Techniques: Produced, Expressing, Ex Vivo, MANN-WHITNEY, Infection, Flow Cytometry, Control, Comparison, Isolation

Journal: bioRxiv

Article Title: Leishmania major co-opts IL-7 feedback in monocytes to suppress CD4⁺ T-cell immunity

doi: 10.64898/2026.03.24.713728

Figure Lengend Snippet: A. Hypothesis (box) and experimental setup for testing the effects of anti-IL-7/IL-7R treatment during L. major infection. Treatment was applied twice (day 16, 20, black arrows, data shown in panel B-F) or once (day 18, grey arrow, data shown in panel C). B-C. Limiting dilution assay (LDA) of L. major tissue burden in the ear following anti-IL-7/IL-7R treatment normalized to isotype control treated animals (B) and L. major infected cell fraction among recruited monocytes with two-dosage (left panel) or single-dosage anti-IL-7/IL-7R treatment (right panel) (C). D. IFNγ + fraction (left) and IFNγ MFI (normalized to isotype control treatment, right) among effector CD4 + T cells isolated from the ears at week 3 p.i. of either isotype antibody (black symbols) or anti-IL-7R/IL-7 treated animals (blue symbols). E. H2KB + and PD-L1 + fraction among fibroblasts, and the MFI of H2KB or PD-L1 among fibroblasts (normalized to isotype control animals), either isotype control (black symbols) or anti-IL-7R/IL-7 treated (blue symbols). F. iNOS + fraction and iNOS MFI (normalized to isotype control animals) among total recruited monocyte-derived cells (left panel) and infected monocyte-derived cells (right panel), either isotype control (black symbols) or anti-IL-7R/IL-7 treated samples (blue symbols). Each symbol in (B-F) represents one individual mouse ear, data pooled from two independent experiments. Horizontal lines denote the mean.. **, p<0.01; *, p<0.05 according to unpaired t-test. G . Experimental setup to determine the long-term effects of anti-IL-7/IL-7R treatment. H. Infection-associated immune cell recruitment determined by ear diameter of isotype control-treated (grey) and anti-IL-7/IL-7R treated (blue) animals. Data were normalized to the mean of the corresponding values of the same ear measured before the treatment. Bars denote the mean+SD. Each symbol represents one individual ear repeatedly measured throughout the infection time course. Data obtained from two independent experiments. ****, p<0.0001 *, p<0.05 according to 2-way ANOVA (treatment, experiment) with Bonferroni post-test. I. Limiting dilution assay (LDA) of L. major tissue burden in the ear at week 7 p.i. of animals previously treated (week 3 p.i.) with isotype control-treated (black symbols) or anti-IL-7/IL-7R treated (blue symbols). J. IFNγ + fraction among CD4 + T cells isolated from the draining lymph node at week 7 p.i. of animals previously treated (week 3 p.i.) with either isotype control (black symbols) or anti-IL-7R/IL-7 antibody (blue symbols). Each symbol in (I-J) represents one individual ear. Data pooled from two independent experiments and normalized to control animals of each experiment for limiting dilution. Horizontal lines denote the median. *, p<0.05 according unpaired Mann-Whitney test. Each symbol represents one individual mouse ear. Horizontal lines denote the mean. ***, p<0.001; according to unpaired Mann-Whitney test.

Article Snippet: The following TaqMan® Gene Expression Assays (Thermo Fisher Scientific) were used according to the manufacturer’s instructions: Il7 (FAM-MGB probe Mm01295804 m1) and Hprt (FAM-MGB probe Mm00446968 m1).

Techniques: Infection, Limiting Dilution Assay, Control, Isolation, Derivative Assay, MANN-WHITNEY