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Editorial
. 2018 Mar;15(3):289-291.
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

Hussein Issaoui  1 , Nour Ghazzaui  1 , Alexis Saintamand  1 , Claire Carrion  1 , Christelle Oblet  1 , Yves Denizot  1
Affiliations
Editorial

The IgH 3' regulatory region super-enhancer does not control IgA class switch recombination in the B1 lineage

Hussein Issaoui et al. Cell Mol Immunol. 2018 Mar.
No abstract available

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
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.
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