Editorial
doi: 10.1038/cmi.2017.103.
Epub 2017 Oct 9.
The IgH 3' regulatory region super-enhancer does not control IgA class switch recombination in the B1 lineage
Affiliations
- PMID: 28990584
- PMCID: PMC5843615
- DOI: 10.1038/cmi.2017.103
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Editorial
The IgH 3' regulatory region super-enhancer does not control IgA class switch recombination in the B1 lineage
Cell Mol Immunol.
2018 Mar.
No abstract available
Conflict of interest statement
The authors declare no conflict of interest.
Figures
3′RR and B1 B-cell IgA CSR. (a) Flow cytometry analysis of IgA+ B1 B-cells in the peritoneal cavities of 3′RR-deficient mice and wt mice in response to pristane. Mice were treated with 1 ml of pristane for 2 months prior to experiments. Cells were washed and counted and 2 × 106 cells were incubated with anti-B220-BV510, anti-IgD-BV421, anti-CD23-PC7, anti-IgM-PE, anti-CD11b-eF780 and anti-IgA-FITC antibodies (Southern Biotechnologies, Birmingham, AL, USA and Beckton Dickinson, Franklin Lakes, NJ, USA) and analyzed using a Fortessa LSR2 (Beckton Dickinson). Left: A representative experiment is shown. Cells were gated on B220lowIgDlowCD23−CD11b+/low cells. Right: Percentages of IgA+ B1 B-cells in seven 3′RR-deficient mice and nine wt mice. The results are reported as mean as the means±SEMs. Significance was determined using the Mann–Whitney U-test. (b) In vitro CSR toward IgA in peritoneal cavity B-cells from 3′RR-deficient mice and wt mice. Same mice as in part (a). Single-cell suspensions of CD43− peritoneal cells were cultured for 4 days at 1 × 106 cells per ml in RPMI 1640 with 10% fetal calf serum, 5 μg/ml LPS, 5 ng/ml TGFβ1 and 5 ng/ml BAFF (PeproTech, Rocky Hill, NJ, USA). Single-cell suspensions of cultured B-cells were incubated with anti-B220-BV510, anti-CD19-PE and anti-IgA-FITC labeled antibodies (Southern Biotechnologies) and analyzed using a Fortessa LSR2 (Beckton Dickinson). Left: A representative experiment is shown. Right: Percentages of IgA+ B1 B-cells in eight 3′RR-deficient mice and eight wt mice. The results are reported as the means±SEMs. (c) IgA levels in the peritoneal cavity of pristane-treated 3′RR-deficient mice and wt mice. Same mice as in part A. ELISAs were performed as described. Results are reported as the means±s.e.m for eleven 3′RR-deficient mice and eight wt mice. (d) IgA levels in culture supernatants of IgA+ B1 B-cells. Same mice as in part B. ELISAs were performed as described. Results are reported as the means±SEMs for twelve 3′RR-deficient mice and eleven wt mice. (e) q-PCR analysis of Iμ–Cα transcripts in cultured IgA+ B1 B-cells from 3′RR-deficient mice and wt mice. q-PCR analyses of Iμ–Cα transcripts were performed as previously described. Same mice as in part (b). The results are reported as the means±s.e.m. of five 3′RR-deficient mice and four wt mice. (f) Representative immunostaining (× 200 magnitude) with antibodies against IgA in the guts of 3′RR-deficient mice (right) and wt mice (left). A representative experiment out of three mice per genotype is shown. (g) Representative immunostaining (× 200 magnitude) with antibodies against B220 in the guts of 3′RR-deficient mice (right) and wt mice (left). A representative experiment out of three mice per genotype is shown.
References
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