Journal: Nature communications

Article Title: Sequence variants influencing the regulation of serum IgG subclass levels.

doi: 10.1038/s41467-024-52470-8

Figure Lengend Snippet: Fig. 1 | Frequency of IgG1 allotypes in Iceland and Sweden and association with IgG1 levels. IgG1 allotypes are defined by one or more missense variants. For each allotype, we include the conventional allotype name, the IMGT numbering, and allotype defining amino acids (AA) at given positions within the IgG1 protein (Eu numbering). Below the respective rsID for the single-nucleotide polymorphism is indicated. Underlined AAs indicate changes from the top allotype, G1m(za). We used phased haplotype data to define the IgG1 allotypes based on the unique presence/absence combination of the missense variants detected in IGHG1 in both Icelanders and Swedes. Single nucleotide polymorphisms (SNPs) corresponding to the underlined AA changes are shown in red. Positions of the missense variants are given in the humanreference genome build Hg38 (a). Frequency (%) of the different allotypes found in Icelandersand Swedes (b).Schematic representation of antibody

Article Snippet: For IgG binding, cells were incubated at 37 °C with monoclonal human IgG1 (anti-trinitrophenol) and subsequently with secondary PE-conjugated goat anti-human IgG1 F(ab’)2 (Southern Biotech, USA).

Techniques:

Journal: Nature communications

Article Title: Sequence variants influencing the regulation of serum IgG subclass levels.

doi: 10.1038/s41467-024-52470-8

Figure Lengend Snippet: Fig. 2 | Deletion of the IGHG1 gene associates with increased IgG3 levels in serum. a A coverage plot showing the rare deletion of the IGHG1 gene that is tagged by the lead variant, rs587597004. Six individuals with different status of the ~28 kb deletion at Hg38-chr14:105734756–105762914 are shown in the figure where each row represents the coverage for an individual. The two indivi- duals on the top are homozygous for the deletion, next two are heterozygous and the bottom two are non-carriers of the deletion (b) Box plots showing abso- lute serum levels of each IgG subclass, stratified on IGHG1 deletion genotype. c Normalized expression of each IgG subclass gene mRNA in whole blood, stra- tified on IGHG1 deletion genotype, showing effects consistent with the serum protein levels. d A graph showing the median RNA-sequence coverage of the IGHG1 gene region in whole blood, stratified by the IGHG1 deletion alleles. RNA sequencing data was available for one homozygous deletion carrier, showing no IGHG1 mRNA. In the box plots, the bottom and top of the boxes correspond to the 25th (Q1) and 75th (Q3) percentiles, the line inside the box is the median, and the whiskers are located at Q1 – 1.5 IQR and Q3 + 1.5 IQR (where IQR is the interquartile range, Q1–Q3). b, c n represents number of carriers for the rare deletion where WT wild-type/non-carriers, Het heterozygous carriers and Hom homozygous carriers.

Article Snippet: For IgG binding, cells were incubated at 37 °C with monoclonal human IgG1 (anti-trinitrophenol) and subsequently with secondary PE-conjugated goat anti-human IgG1 F(ab’)2 (Southern Biotech, USA).

Techniques: Variant Assay, Expressing, Sequencing, RNA Sequencing

Journal: bioRxiv

Article Title: Subcapsular sinus macrophage sensing of extracellular matrix rigidity alters membrane topography and immune complex mobility

doi: 10.1101/2022.12.02.518873

Figure Lengend Snippet: Pseudocolour flow cytometry plots of live single-cell suspensions from (A) whole lymph nodes, and (B,C) enzyme-digested lymph nodes positively enriched by (B) an FDC-specific antibody (rat IgG2c, κ anti-mouse FDC-M1) or (C) a rat IgG2c, κ isotype control antibody, both complexed with a biotinylated mouse IgG2a anti-rat Ig κ secondary antibody and captured by anti-biotin microbeads. Cells were stained with monoclonal antibodies to CD45, CD11b, CD11c, CD169, and F4/80 (see ). The proportions of all cell populations are indicated on the plots. Data are representative of three experiments.

Article Snippet: Immune complexes were generated by mixing 10 μl of the mouse serum, as a source of complement, with 0.5 μg Cy3B-labelled goat IgG1 anti-mouse Ig κ (Southern Biotech; labelled in-house), 0.375 μg donkey anti-goat IgG (H+L) (Jackson ImmunoResearch), and 40 μl of GVB++ buffer (Complement Technology) for 30 minutes at 37 °C.

Techniques: Flow Cytometry, Staining

Journal: bioRxiv

Article Title: Subcapsular sinus macrophage sensing of extracellular matrix rigidity alters membrane topography and immune complex mobility

doi: 10.1101/2022.12.02.518873

Figure Lengend Snippet: (A) Immunofluorescence staining with anti-CD16/32 (Fc γ RIII/Fc γ RII). Both SSMs (F-actin + CD16/32 + ) and non-SSMs (F-actin + CD16/32 - ) are visible in this image. (B, C) The ability of SSMs to capture and present immune complexes does not depend upon complement. IgG antibody complexes are captured equally well when (B) they are (+) and (C) are not (-) fixed with complement. Scale bars: 10 μm.

Article Snippet: Immune complexes were generated by mixing 10 μl of the mouse serum, as a source of complement, with 0.5 μg Cy3B-labelled goat IgG1 anti-mouse Ig κ (Southern Biotech; labelled in-house), 0.375 μg donkey anti-goat IgG (H+L) (Jackson ImmunoResearch), and 40 μl of GVB++ buffer (Complement Technology) for 30 minutes at 37 °C.

Techniques: Immunofluorescence, Staining