(A) Experimental design: C57Bl/6 mice were either left naive or underwent sham or CLP surgery. All groups were immunized s.c. with MOG35-55 emulsified in complete Freunds adjuvant (CFA) on both …
Source data for Figure 1B–D.
Source data for Figure 1E.
Source data for Figure 1F,G.
(A) Experimental design: C57Bl/6 mice were either left naive or underwent sham or CLP surgery. Both groups were immunized s.c. with MOG35-55 emulsified in complete Freunds adjuvant (CFA) on both …
Source data for Figure 1—figure supplement 1.
Sham and CLP mice were immunized s.c. with MOG35-55 emulsified in complete Freunds adjuvant (CFA) on both flanks 5 days after surgery and given pertussis toxin (PTX) i.p. on the day of immunization …
Source data for Figure 1—figure supplement 2.
Sham and CLP mice were immunized s.c. with MOG35-55 emulsified in complete Freunds adjuvant (CFA) on both flanks 5 days after surgery and given pertussis toxin (PTX) i.p. on the day of immunization …
Source data for Figure 1—figure supplement 2.
(A) Experimental design: C57Bl/6 mice underwent either sham or CLP surgery. EAE induction occurred 5 days after surgery. Mice were perfused 15 days post-EAE induction and CNS was harvested. (B) …
Source data for Figure 2.
(A ) Representative flow plots of direct ex vivo IFNγ, IL-17A, and TNFα producing cells, gated on MOG-specific CD4 T cells, from sham (left) and CLP (right) mice. Numbers indicate frequency of …
Source data for Figure 3.
(A) Representative flow plots of direct ex vivo IFNγ, IL-17A, and TNFα producing cells, gated on MOG-specific CD4 T cells, from sham (black) mice. Gray profiles indicate ‘fluorescence minus one’ …
(A) Experimental design: C57Bl/6 mice underwent sham or CLP surgery. EAE induction occurred 5 days after surgery. Draining inguinal lymph nodes (iLN) were harvested 7 days after EAE disease …
Source data for Figure 4.
(A) Experimental design: C57Bl/6 mice underwent sham or CLP surgery. MOG-specific CD4 T cell precursors were enriched from the spleen 5 days after surgery via tetramer pulldown and enumerated. (B) …
Source data for Figure 5.
(A) Experimental design: Thy1.2 C57Bl/6 mice were separated into Pre- and Post-transfer groups. The Pre-transfer group received 5 × 103 naive Thy1.1 2D2 TCR-Tg CD4 T cells 1 day before sham or CLP …
Source data for Figure 6B–D.
Source data for Figure 6B,E.
Thy1.2 C57Bl/6 mice were separated into Pre- and Post-transfer groups. The Pre-transfer group received 5 × 103 naive Thy1.1 2D2 TCR-Tg CD4 T cells 1 day before sham or CLP surgery. The Post-transfer …
Source data for Figure 6—figure supplement 1.
Thy1.2 C57Bl/6 mice were separated into Pre- and Post-transfer groups. The Pre-transfer group received 5 × 103 naive Thy1.1 2D2 TCR-Tg CD4 T cells 1 day before sham or CLP surgery. The Post-transfer …
Source data for Figure 6—figure supplement 2.
(A) Experimental design: Thy1.1/1.1 2D2 TCR-Tg mice underwent sham surgery while Thy1.1/1.2 2D2 TCR-Tg mice underwent CLP surgery. 2D2 mice were euthanized 4 days post-surgery and splenic 2D2 cells …
Source data for Figure 7.
(A) Experimental design: Thy1.2 C57Bl/6 mice were separated into Pre- and Post-transfer groups. The Pre-transfer group received 5 × 103 naive Thy1.1 2D2 TCR-Tg CD4 T cells 1 day before sham or CLP …
Source data for Figure 8.
Source data for Figure 8.
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Strain, strain background (Mus musculus) | C57BL6/J | Jackson Laboratory | Stock No: 000664 (RRID:IMSR_JAX:000664) | |
Strain, strain background (Mus musculus) | B6.PL(84NS)/Cy | Jackson Laboratory | Stock No: 000983 (RRID:IMSR_JAX:000406) | C57BL6/J Thy1.1 |
Strain, strain background (Mus musculus) | C57BL/6-Tg(Tcra2D2,Tcrb2D2)1Kuch/J | Jackson Laboratory | Stock No: 006912 (RRID:IMSR_JAX:006912) | |
Strain, strain background (Mus musculus) | Thy1.1/1.1- C57BL/6-Tg(Tcra2D2,Tcrb2D2)1Kuch/J | This paper | Thy1.1/1.1 2D2 | Can be acquired through lab contact or breeding of above commercially available strains |
Strain, strain background (Mus musculus) | Thy1.1/1.2- C57BL/6-Tg(Tcra2D2,Tcrb2D2)1Kuch/J | This paper | Thy1.1/1.2 2D2 | Can be acquired through lab contact or breeding of above commercially available strains |
Peptide, recombinant protein | MOG35-55 | GenScript | SC1208 | |
Other | CFA containing M. tuberculosis H37Ra | Difco | DF3114-33-8 | |
Peptide, recombinant protein | Pertussis toxin from Bordetella pertussis | Sigma-Aldrich | P7208 | |
Antibody | CD4 (Rat monoclonal) | Biolegend | GK1.5 (AB_312689) | FACs (1:400) |
Antibody | CD11a (Rat monoclonal) | Biolegend | M17/4 (AB_312776) | FACs (1:300) |
Antibody | IFNγ (Rat monoclonal) | eBioscience | XMG1.2 (AB_465410) | FACs (1:100) |
Antibody | IL-17A (Rat monoclonal) | eBioscience | eBio17B7 (AB_906240) | FACs (1:100) |
Antibody | TNFα (Rat monoclonal) | eBioscience | MP6-XT22 (AB_465416) | FACs (1:100) |
Antibody | CD8a (Rat monoclonal) | Biolegend | 5H10-1 (AB_312762) | FACs (1:400) |
Antibody | Ki67 (Mouse monoclonal) | BD Pharmingen | B56 (AB_2858243) | FACs (1:100) |
Antibody | Thy1.1 (Mouse monoclonal) | eBioscience | HIS51 (AB_1257173) | FACs (1:1000) |
Antibody | Thy1.2 (Rat monoclonal) | eBioscience | 53–2.1 (AB_467378) | FACs (1:1000) |
Antibody | CD44 (Rat monoclonal) | eBioscience | IM7 (AB_469715) | FACs (1:200) |
Antibody | CD45 (Mouse monoclonal) | eBioscience | 104 (AB_469724) | FACs (1:100) |
Antibody | F4/80 (Rat monoclonal) | Biolegend | BM8 (AB_893499) | FACs (1:100) |
Antibody | CD11b (Rat monoclonal) | eBioscience | M1/70 (AB_468883) | FACs (1:200) |
Antibody | IA-b (Rat monoclonal) | eBioscience | M5/114.15.2 (AB_529608) | FACs (1:100) |
Antibody | CD3e (Armenian Hamster monoclonal) | eBioscience | 145–2 C11 (AB_467048) | FACs (1:100) |
Antibody | CD19 (Mouse monoclonal) | eBioscience | MB19-1 (AB_467145) | FACs (1:100) |
Antibody | FAS (Mouse monoclonal) | Biolegend | SA367H8 (AB_2629777) | FACs (1:100) |
Antibody | FASL (Armenian Hamster monoclonal) | Biolegend | MFL3 (AB_313276) | FACs (1:100) |
Antibody | TRAIL (Rat monoclonal) | Biolegend | N2B2 (AB_345271) | FACs (1:100) |
Antibody | RORγT (Rat monoclonal) | eBioscience | AFKJS-9 (AB_1834470) | FACs (1:100) |
Antibody | Tbet (Mouse monoclonal) | eBioscience | eBio4b10 (AB_763636) | FACs (1:100) |
Antibody | FoxP3 (Rat monoclonal) | Invitrogen | FJK-16S (AB_467576) | FACs (1:100) |
Antibody | CD11c (Armenian Hamster monoclonal) | Biolegend | N418 (AB_313772) | FACs (1:100) |
Antibody | B220 (Rat monoclonal) | Biolegend | RA3-6B2 (AB_312989) | FACs (1:100) |
Peptide, recombinant protein | MOG40-48 I-Ab linked (Drosophila melanogaster S2 cells) | NIH tetramer core | FACs (1:100) | |
Commercial assay or kit | Foxp3 / Transcription Factor Staining Buffer Set | Invitrogen | 00-5523-00 | |
Commercial assay or kit | Vybrant FAM Caspase-3 and −7 assay kit | Thermo-Fischer | V35118 | |
Commercial assay or kit | BioRad Bio-plex Pro Mouse Cytokine 23-plex | Biorad | M60009RDPD | |
Software, algorithm | GraphPad Prism | GraphPad Prism 8 | Version 8.4.2 (464) (RRID:SCR_002798) |