Human Fcγ-receptor IIb modulates pathogen-specific versus self-reactive antibody responses in lyme arthritis
- Institute of Genetics, Department of Biology, University of Erlangen-Nuremberg, Germany
- Institute of Medical Microbiology and Hygiene, University Regensburg, Germany
- Laboratory for Cancer Immunotherapy, University Hospital Basel, Switzerland
- Department of Dermatology, Laboratory of Dendritic Cell Biology, University Hospital Erlangen, Germany
- Medical Immunology Campus Erlangen, University of Erlangen-Nuremberg, Germany
- Medical Department 3, University Hospital Erlangen, Germany
Figures
Figure 1
The human immune system controls B. burgdorferi infection.
(A, B) Humanized and non-humanized mice were infected with B. burgdorferi and followed for signs of joint inflammation and pathogen spread. In (A) representative pictures of the hind limbs of …
Figure 2 with 2 supplements
Effect of B. burgdorferi infection on the human immune system in humanized mice.
Shown is the effect of B. burgdorferi infection on the abundance (percentage of all human CD45+ cells) and activation status (CD69 expression) of human CD19+ B cells (A-C), CD33+ myeloid cells (D-F),…
Figure 2—figure supplement 1
Gating strategy to identify mouse and human immune cells in humanized mice.
(A) Shown is a representative gating strategy used to identify human immune cells in the peripheral blood of humanized mice. After excluding cell duplets, hCD45+ DAPI negative cells were pre-gated …
Figure 2—figure supplement 2
Infiltration of mouse immune cells in B. burgdorferi infected humanized mice.
(A) Shown are box and whisker plots including individual data points representing the relative abundance of mouse eosinophils, neutrophils (Ly6G+), Ly6C+ monocytes (Ly6C+ mono.), and Ly6C-negative …
Figure 3 with 2 supplements
The role of B cells in B. burgdorferi control.
(A) Schematic representation of the experimental strategy. B cells were depleted with a human CD20-specific antibody (α-CD20) injected 2 days before (d-2), at the day of infection (d0) and 4 and 7 …
Figure 3—figure supplement 1
Effect of human FcγRIIb alleles, T cells and B cells on B. burgdorferi load and on pathogen-specific immune responses.
(A) Shown is an ethidium bromide stained agarose gel electrophoresis of a B. burgdorferi flagellin-specific nested PCR performed on ear tissue of humanized mice carrying the functional (232I, lanes …
Figure 3—figure supplement 2
FcγRIIb expression on dendritic cells and effect of human T cells on B. burgdorferi specific immune responses.
(A) Shown is a representative example of a FACS analysis detecting FcγRIIb expression on human dendritic cells in the spleen of humanized mice using CD45, CD33, CD11c, CD4, and FcγRIIb (using the …
Figure 4
Impact of human FcγRIIb alleles on the humoral immune response and pathogen load during B. burgdorferi infection.
(A) Shown are representative dot plots of CD19+CD138+ plasma blasts (marked by red squares including the respective percentage of CD19+CD138+ cells within the population of CD19+ B cells) in the …
Figure 5
Correlation of human autoreactive and pathogen-specific antibody responses in humanized mice.
Humanized mice homozygous for the functional (FcγRIIb-232I; n = 9) (A) and non-functional (FcγRIIb-232T; n = 12) (B) FcγRIIb alleles were infected with B. burgdorferi and serum samples were …
Figure 6
Induction of B. burgdorferi-specific and autoreactive antibody responses in humans.
Shown is the correlation of serum IgM (µg/ml), glucose 6-phosphate (OD), and DNA specific (OD) IgM responses with B. burgdorferi specific IgM responses in healthy donors (n = 11) (A) and a cohort of …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Antibody | anti-human CD3 PE/Cy7 (mouse monoclonal) | BioLegend | Cat.#:300316 | FACS (1:2000) |
| Antibody | anti-human CD4 APC (rat monoclonal) | BioLegend | Cat.#:357408 | FACS (1:200) |
| Antibody | anti-human CD8a FITC (mouse monoclonal) | BioLegend | Cat.#:301050 | FACS (1:100) |
| Antibody | Anti-human CD11c FITC (mouse monoclonal) | BioLegend | Cat.#:301604 | FACS (1:50) |
| Antibody | Anti-human FcγRIIb PE (recombinant human IgG1-N297A) | in house | Clone 2B6 | FACS (1:50) |
| Antibody | anti-human CD19 Brilliant Violet 510 (mouse monoclonal) | BioLegend | Cat.#:302242 | FACS (1:100) |
| Antibody | anti-human CD20 FITC (mouse monoclonal) | BioLegend | Cat.#:302304 | FACS (1:200) |
| Antibody | anti-human CD33 PE (mouse monoclonal) | BioLegend | Cat.#:303404 | FACS (1:200) |
| Antibody | anti-human CD33 BV510 (mouse monoclonal) | BioLegend | Cat.#:303422 | FACS (1:100) |
| Antibody | anti-human CD45 APC/Fire750 (mouse monoclonal) | BioLegend | Cat.#:304062 | FACS (1:400) |
| Antibody | anti-human CD69 PerCP/Cy5.5 (mouse monoclonal) | BioLegend | Cat.#:310926 | FACS (1:100) |
| Antibody | anti-human CD138 PE/Cy7 (mouse monoclonal) | BioLegend | Cat.#:356514 | FACS (1:200) |
| Antibody | anti-mouse/human CD11b PerCP/Cy5.5 (rat monoclonal) | BioLegend | Cat.#:101228 | FACS (1:1000) |
| Antibody | anti-mouse CD45.1 PE (mouse monoclonal) | BioLegend | Cat.#:110707 | FACS (1:600) |
| Antibody | anti-mouse CD45.1 Brilliant Violet 421 (mouse monoclonal) | BioLegend | Cat.#:110732 | FACS (1:400) |
| Antibody | anti-mouse CD62L PE/Cy7 (rat monoclonal) | BioLegend | Cat.#:104418 | FACS (1:1000) |
| Antibody | anti-mouse F4/80 APC/Fire 750 (rat monoclonal) | BioLegend | Cat.#:123152 | FACS (1:200) |
| Antibody | anti-mouse Gr-1 Brilliant Violet 510 (rat monoclonal) | BioLegend | Cat.#:108437 | FACS (1:300) |
| Antibody | anti-mouse Ly-6G FITC (rat monoclonal) | BioLegend | Cat.#:127606 | FACS (1:300) |
| Antibody | 9E9 Alexa647 (rabbit monoclonal) | Nimmerjahn et al., 2005 | n/a | FACS (1:200) |
| Antibody | Anti-human CD4 (for depletion) (mouse monoclonal) | BioXCell | Cat.#:BE0003-2 | 100 µg per injection |
| Antibody | Anti-human CD20 (for depletion) (recombinant mouse monoclonal) | Kao et al., 2015 | In house | 25 µg per injection |
| Strain, strain background Mus musculus | NOD.Cg-Prkdc<scid > Il2rgtm1Wjl/Szj | The Jackson Laboratory | Cat.#:005557 | n/a |
| Software, algorithm | Graph Pad Prism 7.03 | GraphPad Software Inc, San Diego, CA, USA |
Additional files
-
Source data 1
Summarized source data for all figures.
- https://cdn.elifesciences.org/articles/55319/elife-55319-data1-v2.xlsx
-
Transparent reporting form
- https://cdn.elifesciences.org/articles/55319/elife-55319-transrepform-v2.docx
