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The Extracellular Domains of IgG1 and T Cell-Derived IL-4/IL-13 Are Critical for the Polyclonal Memory IgE Response In Vivo.

Turqueti-Neves A, Otte M, Schwartz C, Schmitt ME, Lindner C, Pabst O, Yu P, Voehringer D - PLoS Biol. (2015)

Bottom Line: The development and persistence of IgE responses are poorly understood, which is in part due to the low number of IgE-producing cells.Together, our results reveal a close relationship between the IgE and IgG1 repertoires in vivo and demonstrate that the memory IgE response is mainly conserved at the level of memory IgG1+ B cells.Therefore, targeting the generation and survival of allergen-specific IgG1+ B cells could lead to development of new therapeutic strategies to treat chronic allergic disorders.

View Article: PubMed Central - PubMed

Affiliation: Department of Infection Biology, Institute for Clinical Microbiology, Immunology and Hygiene, University Hospital Erlangen and Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany.

ABSTRACT
IgE-mediated activation of mast cells and basophils contributes to protective immunity against helminths but also causes allergic responses. The development and persistence of IgE responses are poorly understood, which is in part due to the low number of IgE-producing cells. Here, we used next generation sequencing to uncover a striking overlap between the IgE and IgG1 repertoires in helminth-infected or OVA/alum-immunized wild-type BALB/c mice. The memory IgE response after secondary infection induced a strong increase of IgE+ plasma cells in spleen and lymph nodes. In contrast, germinal center B cells did not increase during secondary infection. Unexpectedly, the memory IgE response was lost in mice where the extracellular part of IgG1 had been replaced with IgE sequences. Adoptive transfer studies revealed that IgG1+ B cells were required and sufficient to constitute the memory IgE response in recipient mice. T cell-derived IL-4/IL-13 was required for the memory IgE response but not for expansion of B cells from memory mice. Together, our results reveal a close relationship between the IgE and IgG1 repertoires in vivo and demonstrate that the memory IgE response is mainly conserved at the level of memory IgG1+ B cells. Therefore, targeting the generation and survival of allergen-specific IgG1+ B cells could lead to development of new therapeutic strategies to treat chronic allergic disorders.

No MeSH data available.


Related in: MedlinePlus

IgG1+ precursors are required for constitution of the memory IgE response.(A) GC B cells (B220+CD38loGL-7+; left plots) and intracellular IgE+ or IgG1+ expression on gated GC B cells (right plots) in mesenteric LN samples from day 14 N. brasiliensis-infected IgEki/ki and wild-type C57BL/6 mice. (B) PCs (B220−CD138+; left plots) were gated from blasts (FSChiSSChi), and percentage of intracellular IgE+ and IgG1+ PCs is shown in the right plots. (C) Serum IgE in IgEki/ki and C57BL/6 mice after first and secondary N. brasiliensis infection. (D) Ratio of IgEb to IgEa in IgEki(a)/wt(b) or IgEwt(a)/wt(b) mice after first and secondary N. brasiliensis infection. (E) 1.5 x 105 sorted B220+IgM−IgD−IgE−, B220+IgM−IgD−IgG1−, B220+IgG1+ B cells or IgG1−CD138+ PCs gated as indicated in S7 Fig from IgHa mice on d13 after secondary N. brasiliensis infection were transferred into nonirradiated IgHb mice. Recipient mice were infected with N. brasiliensis and IgEa was determined in the serum on day 12 after infection. Data show the mean + SEM from four independent experiments and at least seven mice (A and B), two experiments with eight mice total per group (C), two experiment with 2–7 mice (E). *p < 0.05, ***p < 0.001 by Student’s t test. n. d. = not detectable.
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pbio.1002290.g009: IgG1+ precursors are required for constitution of the memory IgE response.(A) GC B cells (B220+CD38loGL-7+; left plots) and intracellular IgE+ or IgG1+ expression on gated GC B cells (right plots) in mesenteric LN samples from day 14 N. brasiliensis-infected IgEki/ki and wild-type C57BL/6 mice. (B) PCs (B220−CD138+; left plots) were gated from blasts (FSChiSSChi), and percentage of intracellular IgE+ and IgG1+ PCs is shown in the right plots. (C) Serum IgE in IgEki/ki and C57BL/6 mice after first and secondary N. brasiliensis infection. (D) Ratio of IgEb to IgEa in IgEki(a)/wt(b) or IgEwt(a)/wt(b) mice after first and secondary N. brasiliensis infection. (E) 1.5 x 105 sorted B220+IgM−IgD−IgE−, B220+IgM−IgD−IgG1−, B220+IgG1+ B cells or IgG1−CD138+ PCs gated as indicated in S7 Fig from IgHa mice on d13 after secondary N. brasiliensis infection were transferred into nonirradiated IgHb mice. Recipient mice were infected with N. brasiliensis and IgEa was determined in the serum on day 12 after infection. Data show the mean + SEM from four independent experiments and at least seven mice (A and B), two experiments with eight mice total per group (C), two experiment with 2–7 mice (E). *p < 0.05, ***p < 0.001 by Student’s t test. n. d. = not detectable.

Mentions: We performed further experiments to clarify whether the memory IgE response is dependent on an IgG1+ memory B cell. Here, we used the recently described IgE knock-in mouse (IgEki/ki) in which the extracellular part of the IgG1 heavy chain had been replaced by the extracellular part of the IgE heavy chain [36]. When mesenteric LN were analyzed on day 14 after N. brasiliensis infection, wild-type mice had about 43% IgG1+ and 1.6% IgE+ GC B cells, while IgEki/ki mice had no IgG1+ and about 27% IgE+ GC B cells (Fig 9A). However, this 20-fold increased population of IgE+ GC B cells in IgEki/ki mice did not result in the same increase of IgE+ PCs (Fig 9B), and the serum IgE levels were only about 2-fold higher in IgEki/ki mice as compared to control mice at the peak of the primary response to N. brasiliensis (Fig 9C). Interestingly, serum IgE levels during the recall response in IgEki/ki mice were comparable to the IgE levels after primary infection and much lower compared to IgE levels in control mice (Fig 9C). We further analyzed the IgE response in F1 mice generated by crossing IgEki/ki mice (IgHa) or normal IgHa mice to C57BL/6 mice (IgHb). Due to allelic exclusion, about 50% of B cells in these mice express the IgHa allele while the other 50% express the IgHb allele. We found that IgEb dominated the memory IgE response to N. brasiliensis in IgEki(a)/wt(b) mice illustrating the competitive advantage of the wild-type allele encoding the extracellular domains of IgG1 (Fig 9D). Next, we sorted different B cell subsets and PCs from N. brasiliensis-infected IgHa memory mice and transferred them separately into naïve nonirradiated IgHb recipient mice. After N. brasiliensis infection of recipient mice, we observed a prominent IgE response in recipients of B220+IgG1+ and B220+IgM−IgD−IgE− cells but not in recipients of B220+IgM−IgD−IgG1− or IgG1− PCs (Fig 9E and S7 Fig).


The Extracellular Domains of IgG1 and T Cell-Derived IL-4/IL-13 Are Critical for the Polyclonal Memory IgE Response In Vivo.

Turqueti-Neves A, Otte M, Schwartz C, Schmitt ME, Lindner C, Pabst O, Yu P, Voehringer D - PLoS Biol. (2015)

IgG1+ precursors are required for constitution of the memory IgE response.(A) GC B cells (B220+CD38loGL-7+; left plots) and intracellular IgE+ or IgG1+ expression on gated GC B cells (right plots) in mesenteric LN samples from day 14 N. brasiliensis-infected IgEki/ki and wild-type C57BL/6 mice. (B) PCs (B220−CD138+; left plots) were gated from blasts (FSChiSSChi), and percentage of intracellular IgE+ and IgG1+ PCs is shown in the right plots. (C) Serum IgE in IgEki/ki and C57BL/6 mice after first and secondary N. brasiliensis infection. (D) Ratio of IgEb to IgEa in IgEki(a)/wt(b) or IgEwt(a)/wt(b) mice after first and secondary N. brasiliensis infection. (E) 1.5 x 105 sorted B220+IgM−IgD−IgE−, B220+IgM−IgD−IgG1−, B220+IgG1+ B cells or IgG1−CD138+ PCs gated as indicated in S7 Fig from IgHa mice on d13 after secondary N. brasiliensis infection were transferred into nonirradiated IgHb mice. Recipient mice were infected with N. brasiliensis and IgEa was determined in the serum on day 12 after infection. Data show the mean + SEM from four independent experiments and at least seven mice (A and B), two experiments with eight mice total per group (C), two experiment with 2–7 mice (E). *p < 0.05, ***p < 0.001 by Student’s t test. n. d. = not detectable.
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Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4629909&req=5

pbio.1002290.g009: IgG1+ precursors are required for constitution of the memory IgE response.(A) GC B cells (B220+CD38loGL-7+; left plots) and intracellular IgE+ or IgG1+ expression on gated GC B cells (right plots) in mesenteric LN samples from day 14 N. brasiliensis-infected IgEki/ki and wild-type C57BL/6 mice. (B) PCs (B220−CD138+; left plots) were gated from blasts (FSChiSSChi), and percentage of intracellular IgE+ and IgG1+ PCs is shown in the right plots. (C) Serum IgE in IgEki/ki and C57BL/6 mice after first and secondary N. brasiliensis infection. (D) Ratio of IgEb to IgEa in IgEki(a)/wt(b) or IgEwt(a)/wt(b) mice after first and secondary N. brasiliensis infection. (E) 1.5 x 105 sorted B220+IgM−IgD−IgE−, B220+IgM−IgD−IgG1−, B220+IgG1+ B cells or IgG1−CD138+ PCs gated as indicated in S7 Fig from IgHa mice on d13 after secondary N. brasiliensis infection were transferred into nonirradiated IgHb mice. Recipient mice were infected with N. brasiliensis and IgEa was determined in the serum on day 12 after infection. Data show the mean + SEM from four independent experiments and at least seven mice (A and B), two experiments with eight mice total per group (C), two experiment with 2–7 mice (E). *p < 0.05, ***p < 0.001 by Student’s t test. n. d. = not detectable.
Mentions: We performed further experiments to clarify whether the memory IgE response is dependent on an IgG1+ memory B cell. Here, we used the recently described IgE knock-in mouse (IgEki/ki) in which the extracellular part of the IgG1 heavy chain had been replaced by the extracellular part of the IgE heavy chain [36]. When mesenteric LN were analyzed on day 14 after N. brasiliensis infection, wild-type mice had about 43% IgG1+ and 1.6% IgE+ GC B cells, while IgEki/ki mice had no IgG1+ and about 27% IgE+ GC B cells (Fig 9A). However, this 20-fold increased population of IgE+ GC B cells in IgEki/ki mice did not result in the same increase of IgE+ PCs (Fig 9B), and the serum IgE levels were only about 2-fold higher in IgEki/ki mice as compared to control mice at the peak of the primary response to N. brasiliensis (Fig 9C). Interestingly, serum IgE levels during the recall response in IgEki/ki mice were comparable to the IgE levels after primary infection and much lower compared to IgE levels in control mice (Fig 9C). We further analyzed the IgE response in F1 mice generated by crossing IgEki/ki mice (IgHa) or normal IgHa mice to C57BL/6 mice (IgHb). Due to allelic exclusion, about 50% of B cells in these mice express the IgHa allele while the other 50% express the IgHb allele. We found that IgEb dominated the memory IgE response to N. brasiliensis in IgEki(a)/wt(b) mice illustrating the competitive advantage of the wild-type allele encoding the extracellular domains of IgG1 (Fig 9D). Next, we sorted different B cell subsets and PCs from N. brasiliensis-infected IgHa memory mice and transferred them separately into naïve nonirradiated IgHb recipient mice. After N. brasiliensis infection of recipient mice, we observed a prominent IgE response in recipients of B220+IgG1+ and B220+IgM−IgD−IgE− cells but not in recipients of B220+IgM−IgD−IgG1− or IgG1− PCs (Fig 9E and S7 Fig).

Bottom Line: The development and persistence of IgE responses are poorly understood, which is in part due to the low number of IgE-producing cells.Together, our results reveal a close relationship between the IgE and IgG1 repertoires in vivo and demonstrate that the memory IgE response is mainly conserved at the level of memory IgG1+ B cells.Therefore, targeting the generation and survival of allergen-specific IgG1+ B cells could lead to development of new therapeutic strategies to treat chronic allergic disorders.

View Article: PubMed Central - PubMed

Affiliation: Department of Infection Biology, Institute for Clinical Microbiology, Immunology and Hygiene, University Hospital Erlangen and Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany.

ABSTRACT
IgE-mediated activation of mast cells and basophils contributes to protective immunity against helminths but also causes allergic responses. The development and persistence of IgE responses are poorly understood, which is in part due to the low number of IgE-producing cells. Here, we used next generation sequencing to uncover a striking overlap between the IgE and IgG1 repertoires in helminth-infected or OVA/alum-immunized wild-type BALB/c mice. The memory IgE response after secondary infection induced a strong increase of IgE+ plasma cells in spleen and lymph nodes. In contrast, germinal center B cells did not increase during secondary infection. Unexpectedly, the memory IgE response was lost in mice where the extracellular part of IgG1 had been replaced with IgE sequences. Adoptive transfer studies revealed that IgG1+ B cells were required and sufficient to constitute the memory IgE response in recipient mice. T cell-derived IL-4/IL-13 was required for the memory IgE response but not for expansion of B cells from memory mice. Together, our results reveal a close relationship between the IgE and IgG1 repertoires in vivo and demonstrate that the memory IgE response is mainly conserved at the level of memory IgG1+ B cells. Therefore, targeting the generation and survival of allergen-specific IgG1+ B cells could lead to development of new therapeutic strategies to treat chronic allergic disorders.

No MeSH data available.


Related in: MedlinePlus