Sepsis impedes EAE disease development and diminishes autoantigen-specific naive CD4 T cells

  1. Isaac J Jensen
  2. Samantha N Jensen
  3. Frances V Sjaastad
  4. Katherine N Gibson-Corley
  5. Thamothrampillai Dileepan
  6. Thomas S Griffith
  7. Ashutosh K Mangalam  Is a corresponding author
  8. Vladimir P Badovinac  Is a corresponding author
  1. Interdisciplinary Graduate Program in Immunology, University of Iowa, United States
  2. Microbiology, Immunology, and Cancer Biology PhD Program, University of Minnesota, United States
  3. Department of Pathology, University of Iowa, Holden Comprehensive Cancer Center, University of Iowa Hospitals and Clinics, United States
  4. Department of Microbiology and Immunology, University of Minnesota, Center for Immunology, United States
  5. Microbiology, Immunology, and Cancer Biology PhD Program, Department of Urology, Center for Immunology, Minneapolis VA Health Care System, University of Minnesota, United States
  6. Interdisciplinary Graduate Program in Immunology, Department of Pathology, University of Iowa, United States
  7. Interdisciplinary Graduate Program in Immunology, Department of Pathology, Department of Microbiology and Immunology, University of Iowa, United States
8 figures, 1 table and 1 additional file

Figures

Figure 1 with 3 supplements
Sepsis ablates EAE disease and reduces CNS pathology.

(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 …

Figure 1—figure supplement 1
Sepsis ablates EAE disease and reduces CNS pathology.

(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 …

Figure 1—figure supplement 2
Altered systemic cytokine response to EAE immunization in septic mice.

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 …

Figure 1—figure supplement 3
Reduced representation and number of activated microglia, monocytes, and macrophages in the CNS of CLP mice following EAE immunization.

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 …

Fewer MOG-specific CD4 T cells are present in the CNS of CLP mice.

(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) …

Figure 3 with 1 supplement
Sepsis reduces the number of cytokine-producing MOG-specific CD4 T cells in the CNS.

(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 …

Figure 3—figure supplement 1
FMO controls for direct ex vivo cytokine staining.

(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’ …

Sepsis reduces the number MOG-specific CD4 T cells present in sites of T cell priming following EAE induction.

(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 …

Sepsis reduces the number of naive MOG-specific CD4 T cell precursors.

(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) …

Figure 6 with 2 supplements
Sepsis reduces the number of MOG-specific CD4 T cells but not their capacity to proliferate.

(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 …

Figure 6—figure supplement 1
Sepsis does not influence the expression of Fas, FasL, and TRAIL by autoantigen-specific T cells.

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 …

Figure 6—figure supplement 2
Sepsis does not influence the expression of RORγT, Tbet, and FoxP3 by autoantigen-specific T cells.

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 …

Sepsis does not lead to a cell intrinsic deficit in CD4 T cell expansion.

(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 …

Sepsis-induced numerical loss of MOG-specific naive CD4 T cell precursors diminishes EAE disease development.

(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 …

Tables

Key resources table
Reagent type
(species) or
resource
DesignationSource or
reference
IdentifiersAdditional
information
Strain, strain background (Mus musculus)C57BL6/JJackson LaboratoryStock No: 000664
(RRID:IMSR_JAX:000664)
Strain, strain background (Mus musculus)B6.PL(84NS)/CyJackson LaboratoryStock No: 000983
(RRID:IMSR_JAX:000406)
C57BL6/J
Thy1.1
Strain, strain background (Mus musculus)C57BL/6-Tg(Tcra2D2,Tcrb2D2)1Kuch/JJackson LaboratoryStock No: 006912
(RRID:IMSR_JAX:006912)
Strain, strain background (Mus musculus)Thy1.1/1.1- C57BL/6-Tg(Tcra2D2,Tcrb2D2)1Kuch/JThis paperThy1.1/1.1 2D2Can 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/JThis paperThy1.1/1.2 2D2Can be acquired through lab contact or breeding of above commercially available strains
Peptide, recombinant proteinMOG35-55GenScriptSC1208
OtherCFA containing M. tuberculosis H37RaDifcoDF3114-33-8
Peptide, recombinant proteinPertussis toxin from Bordetella pertussisSigma-AldrichP7208
AntibodyCD4
(Rat monoclonal)
BiolegendGK1.5
(AB_312689)
FACs
(1:400)
AntibodyCD11a
(Rat monoclonal)
BiolegendM17/4
(AB_312776)
FACs
(1:300)
AntibodyIFNγ
(Rat monoclonal)
eBioscienceXMG1.2
(AB_465410)
FACs
(1:100)
AntibodyIL-17A
(Rat monoclonal)
eBioscienceeBio17B7
(AB_906240)
FACs
(1:100)
AntibodyTNFα
(Rat monoclonal)
eBioscienceMP6-XT22
(AB_465416)
FACs
(1:100)
AntibodyCD8a
(Rat monoclonal)
Biolegend5H10-1
(AB_312762)
FACs
(1:400)
AntibodyKi67
(Mouse monoclonal)
BD PharmingenB56
(AB_2858243)
FACs
(1:100)
AntibodyThy1.1
(Mouse monoclonal)
eBioscienceHIS51
(AB_1257173)
FACs
(1:1000)
AntibodyThy1.2
(Rat monoclonal)
eBioscience53–2.1
(AB_467378)
FACs
(1:1000)
AntibodyCD44
(Rat monoclonal)
eBioscienceIM7
(AB_469715)
FACs
(1:200)
AntibodyCD45
(Mouse monoclonal)
eBioscience104
(AB_469724)
FACs
(1:100)
AntibodyF4/80
(Rat monoclonal)
BiolegendBM8
(AB_893499)
FACs
(1:100)
AntibodyCD11b
(Rat monoclonal)
eBioscienceM1/70
(AB_468883)
FACs
(1:200)
AntibodyIA-b
(Rat monoclonal)
eBioscienceM5/114.15.2
(AB_529608)
FACs
(1:100)
AntibodyCD3e
(Armenian Hamster monoclonal)
eBioscience145–2 C11
(AB_467048)
FACs
(1:100)
AntibodyCD19
(Mouse monoclonal)
eBioscienceMB19-1
(AB_467145)
FACs
(1:100)
AntibodyFAS
(Mouse monoclonal)
BiolegendSA367H8
(AB_2629777)
FACs
(1:100)
AntibodyFASL
(Armenian Hamster monoclonal)
BiolegendMFL3
(AB_313276)
FACs
(1:100)
AntibodyTRAIL
(Rat monoclonal)
BiolegendN2B2
(AB_345271)
FACs
(1:100)
AntibodyRORγT
(Rat monoclonal)
eBioscienceAFKJS-9
(AB_1834470)
FACs
(1:100)
AntibodyTbet
(Mouse monoclonal)
eBioscienceeBio4b10
(AB_763636)
FACs
(1:100)
AntibodyFoxP3
(Rat monoclonal)
InvitrogenFJK-16S
(AB_467576)
FACs
(1:100)
AntibodyCD11c
(Armenian Hamster monoclonal)
BiolegendN418
(AB_313772)
FACs
(1:100)
AntibodyB220
(Rat monoclonal)
BiolegendRA3-6B2
(AB_312989)
FACs
(1:100)
Peptide, recombinant proteinMOG40-48
I-Ab linked
(Drosophila melanogaster S2 cells)
NIH tetramer coreFACs
(1:100)
Commercial assay or kitFoxp3
/
Transcription
Factor
Staining Buffer Set
Invitrogen00-5523-00
Commercial assay or kitVybrant FAM Caspase-3 and −7 assay kitThermo-FischerV35118
Commercial assay or kitBioRad Bio-plex Pro Mouse Cytokine 23-plexBioradM60009RDPD
Software, algorithmGraphPad PrismGraphPad Prism 8Version 8.4.2 (464)
(RRID:SCR_002798)

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