IL-4 and helminth infection downregulate MINCLE-dependent macrophage response to mycobacteria and Th17 adjuvanticity

  1. Judith Schick
  2. Meltem Altunay
  3. Matthew Lacorcia
  4. Nathalie Marschner
  5. Stefanie Westermann
  6. Julia Schluckebier
  7. Christoph Schubart
  8. Barbara Bodendorfer
  9. Dennis Christensen
  10. Christian Alexander
  11. Stefan Wirtz
  12. David Voehringer
  13. Clarissa Prazeres da Costa
  14. Roland Lang  Is a corresponding author
  1. Institut für Klinische Mikrobiologie, Immunologie und Hygiene, Universitätsklinikum Erlangen, Friedrich-Alexander Universität Erlangen-Nürnberg, Germany
  2. Institut für Medizinische Mikrobiologie, Immunologie und Hygiene, Center for Global Health, Technische Universität München, Germany
  3. Center for Global Health, Technical University Munich, Germany
  4. Infektionsbiologische Abteilung, Universitätsklinikum Erlangen, Friedrich-Alexander Universität Erlangen-Nürnberg, Germany
  5. Adjuvant Research, Department of Infectious Disease Immunology, Statens Serum Institut, Denmark
  6. Cellular Microbiology, Forschungszentrum Borstel, Leibniz Lung Center Borstel, Germany
  7. Medizinische Klinik 1, Universitätsklinikum Erlangen, Friedrich-Alexander Universität Erlangen-Nürnberg, Germany
8 figures, 1 table and 1 additional file

Figures

IL-4 impairs upregulation of MINCLE and other DECTIN-2 family C-type lectin receptor (CLR) in macrophages stimulated with bacille Calmette-Guerin (BCG).

(A–D) C57BL/6 bone marrow-derived macrophages (BMM) were stimulated as indicated in presence or absence of IL-4 for 6, 24, or 48 hr. BCG was used at MOI 10. MINCLE (A), DECTIN-2 (B), MCL (C), and …

Figure 1—source data 1

Source data Figure 1 (qRT-PCR and flow cytometry analysis of CLR in macrophages).

https://cdn.elifesciences.org/articles/72923/elife-72923-fig1-data1-v1.xlsx
IL-4 does not affect phagocytosis of bacille Calmette-Guerin (BCG) but inhibits cytokine production.

(A) C57BL/6 bone marrow-derived macrophages (BMMs) were infected with different MOI of fluorescent BCG-Dsred co-treated with IL-4 or not as indicated. Phagocytic uptake was measured via flow …

Overexpression of IL-4 or co-infection with Nippostrongylus brasiliensis impair MINCLE upregulation on peritoneal monocytes but does not reduce phagocytosis upon bacille Calmette-Guerin (BCG) infection.

(A) IL-4 concentration in serum of mice injected with IL-4 minicircle. C57BL/6 mice were hydrodynamically injected with 0.25 or 0.5 µg of IL-4 plasmid (i.v.) or Ringer solution. Five days (0.5 µg) …

Co-infection with Schistosoma mansoni (S.m.) suppresses Th1/Th17 induction by a MINCLE-dependent adjuvant in the spleen but not in the draining lymph node.

(A) Scheme of experimental procedure. Cercariae of S.m. were injected s.c. into C57BL/6 mice. Eight to 9 weeks p.i. mice were immunized with CAF01 (B, C, D) or CpG ODN (E, F). Seven days after …

Co-infection with Nippostrongylus brasiliensis suppresses Th1/Th17 induction by a MINCLE-dependent adjuvant in the spleen but not in the draining lymph node.

(A) Scheme of experimental procedure. C57BL/6 mice were infected subcutaneously in the flank with 500 L3 larvae of N. brasiliensis in 200 µl PBS. Five days p.i. mice were immunized with H1/CAF01 (B–F

Inhibition of Th1/Th17 induction in Nippostrongylus brasiliensis infection depends on IL-4 or IL-13.

C57BL/6 or of Il4-/-Il13-/- mice were infected with N. brasiliensis or not as indicated and immunized s.c. with H1/CAF01 5 days later. On day 7 after immunization, mice were sacrificed and …

Author response image 1
Author response image 2

Tables

Key resources table
Reagent type (species) or resourceDesignationSource or referenceIdentifiersAdditional information
Strain, strain
background
(Mus musculus)
C57BL/6 miceCharles River or Preclinical Experimental
Animal Center
University Hospital Erlangen
Strain, strain
background
(Mus musculus)
MINCLE knockout miceConsortium for Functional GlycomicsClec4e-/-Wells C et al. 2007J
Immunol PMID:18490740
Strain, strain
background
(Mus musculus)
FcRγ knockout miceProf. Dr Falk Nimmerjahn, Erlangen, GermanyFcer1g-/-Takai T et al. 1994Cell PMID:8313472
Strain, strain
background
(Mus musculus)
4-13ko miceProf. AN McKenzie,
Cambridge, UK
Il4-/- Il13-/-McKenzie et al., 1999 J Exp Med PMID:10330435
AntibodyRat monoclonal anti-
mouse Mincle (# 4A9)
Unlabeled
MBLMBL-D292-31:200 dilution
AntibodyRat monoclonal IgG1
Isotype unlabeled
eBioscience17-4812-821:200 dilution
AntibodyRat monoclonal anti-
mouse CD3-APC-eF780
eBioscience47-0031-821:100 dilution
AntibodyRat monoclonal anti-
mouse CD19-APC-eF780
eBioscience47-0193-821:100 dilution
AntibodyRat monoclonal anti-
mouse NK1.1-APC-eF780
eBioscience47-5941-821:100 dilution
AntibodyRat monoclonal anti-
mouse CD11b-FITC
Biolegend1012061:100 dilution
AntibodyRat monoclonal anti-
mouse SiglecF-BV421
BD5626811:400 dilution
AntibodyRat monoclonal anti-
mouse Ly6C-PerCP-Cy5.5
Biolegend1280121:200 dilution
AntibodyRat monoclonal anti-
mouse Ly6G-PE-Cy7
Biolegend1276181:400 dilution
AntibodyHuman monoclonal
anti-mouse Dectin1-APC
Miltenyi130-102-2501:50 dilution
AntibodyHuman monoclonal
anti-mouse Dectin2-APC
Miltenyi130-116-9111:50 dilution
AntibodyHuman IgG1 APC
(Isotype for Dectin2,
Dectin1)
Miltenyi130-113-4461:50 dilution
Chemical
compound, drug
Mouse Fixable
viability dye eF506
eBioscience65-0866-181:1000 dilution
AntibodyRat IgG1 APCeBioscience17-4301-811:200 dilution
AntibodyRat IgG1 APC Iso keBioscience14-4301-821:200 dilution
Sequence-
based reagent
Mincle Primer
(qRT-PCR)
MetabionFor: gctcacctggtggttatcg
Rev: aggttttgtgcgaaaaagga
Sequence-
based reagent
Mcl Primer (qRT-PCR)MetabionFor: agtaacgtgcatccgagagg
Rev: taacaggacagcaggtccaa
Sequence-
based reagent
Dectin2 Primer
(qRT-PCR)
MetabionFor: cagtgaagggactatggtgtca
Rev: ggagccaaatgacttccagt
Sequence-
based reagent
Dectin1 Primer
(qRT-PCR)
MetabionFor: atggttctgggaggatggat
Rev: atggttctgggaggatggat
Sequence-
based reagent
Hprt Primer
(qRT-PCR)
MetabionFor: tcctcctcagaccgctttt
Rev: cctggttcatcatcgctaatc
Sequence-
based reagent
Mincle ProbeRocheRoche Universal Probe Library (UPL)#15
Sequence-
based reagent
Mcl ProbeRocheRoche UPL#1
Sequence-
based reagent
Dectin2 ProbeRocheRoche UPL#89
Sequence-
based reagent
Dectin1 ProbeRocheRoche UPL#60
Sequence-
based reagent
Hprt ProbeRocheRoche UPL#95
Recombinant
DNA reagent
Minicircle DNA encoding IL-4Dr Stefan Wirtz, Erlangen, Germany
Strain, strain
background
(bacteria)
Mycobacterium bovis
BCG DsRed Danish
Dr Anca Dorhoi, Friedrich Löffler Institute, Greifswald, Germany
Strain, strain
background
(helminth)
Nippostrongylus brasiliensisDr David Vöhringer, Erlangen, Germany500 L3 larvae per mouse s.c.
Strain, strain
background
(helminth)
Schistosoma mansoniDr Clarissa Prazeres da Costa, Munich, GermanyNMRI strain100 cercariae per mouse s.c.
OtherCpG ODN1826TIB MOLBIOL1800002370.5 μM (in vitro); 10 nmol
per mouse in vivo
OtherCAF01 (TDB+DDA
liposomes)
Dr Dennis Christensen, Statens Serum Institute, Copenhagen, DenmarkAdjuvant, 50 µl injected
s.c. together with H1 protein
OtherG3D6A liposomal
formulation
Dr
Christian Alexander,
Research Center
Borstel
Adjuvant, 50 µl injected
s.c. together with H1 protein
Peptide,
Recombinant
protein
H1 Mycobacterial
antigen
Dr Dennis Christensen, Statens Serum Institute, Copenhagen, DenmarkH1 is a fusion protein of Ag85B
and ESAT-6.
2 µg H1 in 100 μl
CAF01 for immunization
and 1 μg/ml for re-stimulation
Peptide,
Recombinant
protein
Recombinant murine IL-4PeproTech214–14 B10 μg/ml for in vitro
stimulation
Peptide,
Recombinant
protein
Purified anti-CD3 eBiolegend1003020.5 μg/ml re-stimulation

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