Notícia

Allergy, Asthma & Clinical Immunology (Canadá)

Preconception allergen sensitization can induce B10 cells in offspring: a potential main role for maternal IgG

Publicado em 16 abril 2017

Por Marília Garcia de Oliveira, Luana de Mendonça Oliveira, Aline Aparecida de Lima Lira, Fábio da Ressureição Sgnotto, Alberto José da Silva Duarte, Maria Notomi Sato and Jefferson Russo Victor

Allergy, Asthma & Clinical Immunology

Research

Open Access

Preconception allergen sensitization can induce B10 cells in offspring: a potential main role for maternal IgG

Marília Garcia de Oliveira 1 ,

Luana de Mendonça Oliveira 1 ,

Aline Aparecida de Lima Lira 1 ,

Fábio da Ressureição Sgnotto 1 ,

Alberto José da Silva Duarte 1 , 2 ,

Maria Notomi Sato 1 and

Abstract

Background

The mechanisms through which allergies can be inhibited after preconception immunization with allergens are not fully understood. We aimed to evaluate whether maternal immunization can induce a regulatory B (B10) cell population in offspring in concert with allergy inhibition.

Methods

C57BL/6 females were or were not immunized with OVA and were mated with normal WT males. Their offspring were evaluated at 3 days of age or 20 days after neonatal immunization. Human peripheral B cells from atopic and non-atopic individuals were also evaluated.

Results

Preconception OVA immunization induced B10 cells in offspring, and IL-10 production appeared to be critical for Fc?RIIB upregulation in offspring B cells. Murine and human IL-10-producing B cells responded in vitro to IgG according to the atopic repertoire of the cells.

Conclusions

Our results reveal that maternal immunization induces allergen-specific B10 cells in offspring and a pivotal role for the IgG repertoire in IL-10 production by murine and human B cells.

Keywords

Allergy Allergen-specific IgG B10 cells IL-10 Maternal immunization

Background

Our group has studied type I hypersensitivity inhibition in murine models for the last decade [ 1 – 7 ]. During this period, we proposed that maternal IgG can play a pivotal role in offspring immune modulation, but the mechanisms underlying this phenomenon were not fully elucidated. In the 90s, it was revealed that maternal antibodies are passively transferred to offspring during the gestational and weaning periods. These antibodies, particularly IgG, can interact directly with the offspring’s immune system, even in the absence of antigens [ 8 ], and might involve maternal antibody-allergen immune complexes that directly interact with the inhibitory receptor Fc?RIIb (CD32b) expressed by offspring B cells [ 9 ]. These interactions can in turn regulate the production of IgE antibodies, thus inhibiting the development of allergies. The alterations in the offspring immune system that occur as a consequence of maternal IgG interactions are not fully understood. Because the main event of allergy inhibition is the control of IgE production by B cells, we believe that this population plays a pivotal role as the subject of the effects induced by maternal immunization.

Some populations of B cells can acquire regulatory properties (Breg), and among these cell populations, regulatory B (B10) cells, which have been identified in humans and mice, have high regulatory potential [ 10 ]. This population is characterized by a CD19+IL10+CD1dhigh phenotype and high IL-10 secretion. The regulatory potential of B10 cells can inhibit OVA-induced allergic pulmonary inflammation [ 11 ]. The mechanism through which allergen-specific B10 cells can be induced and a possible role of IgG in this mechanism have not been described.

The aim of this study was to determine whether maternal immunization can induce regulatory B10 cells in offspring as a mechanism of allergy inhibition. Furthermore, we evaluated the importance of maternal IgG for the induction of B10 cells and whether evidence of similar mechanisms can be detected in humans.

Methods

Mice

C57BL/6 inbred wild-type (WT) or IL-10-genetically-deficient male and female mice were used at 8–10 weeks of age. The animals were purchased through the Central Animal Facility of the School of Medicine and Institute of Biomedical Sciences—USP. The offspring (F1) of both sexes were evaluated during the neonatal period, and samples from at least three independent experiments were studied.

Patient samples

Peripheral blood mononuclear cells (PBMCs) and sera were collected from volunteers who were previously classified as atopic or non-atopic individuals according to their clinical status and who voluntarily submitted to a skin prick test (SPT) to confirm their atopic state. These individuals were classified into two groups: atopic individuals [clinically allergic and reactive to at least two allergens, n = 14, age (mean ± SE) = 31.1 ± 2.86] and non-atopic individuals [without any clinical allergy symptoms and unreactive to any tested allergen, n = 14, age (mean ± SE) = 28.2 ± 3.11].

Each PBMC sample was provided by a different donor and analyzed in three independent experiments.

SPT and collection of blood samples

The subjects voluntarily submitted to a SPT according to European standards [ 12 ] with an adapted panel of allergens that included the Brazil pattern (Blomia tropicalis, Canis familiaris, Periplaneta americana, Aspergillus fumigatus, Penicillium notatum, Alternaria alternata, Cladosporium herbarum, Dermatophagoides pteronyssinus, Dermatophagoides farinae, and Felis domesticus).

In brief, a drop of each allergen extract, histamine (positive control) or allergen diluent (negative control—IPI-Asac, Brazil) was applied to the volar aspect of the forearm. A superficial skin puncture was made through each allergen or control drop with a hypodermic lancet (Alko, Brazil) without inducing bleeding. Fifteen minutes after puncture, the transverse diameter of each wheal reaction was measured. We considered a reaction to be positive when the wheal was 3 mm greater than the wheal diameter of the negative control. As exclusion criteria, we adopted the use of antihistamines, glucocorticosteroids and some other systemic drugs that can influence the results 15 days before the test. We also excluded volunteers with severe eczema or dermographism.

Purification of mouse and human IgG

IgG antibodies were purified from the sera of mice that were immunized with OVA (40 days after immunization) or of non-immunized mice using a Melon Gel IgG Spin Purification kit, according to the manufacturer’s instructions (Thermo, USA). In brief, 500 µL of purification gel was placed in a mini column attached to a microtube, and the mixture was centrifuged for 1 min at 2000g. After the supernatant was discarded, 300 µL of washing buffer from the kit was added, and the resulting mixture was centrifuged again. The sample of pooled sera was added to the gel, homogenized for 5 min, and centrifuged, and the supernatant (purified IgG) was collected and stored at -70 °C for subsequent use in culture experiments.

To purify human IgG, two blood samples were obtained by venipuncture from each atopic or non-atopic individual in tubes without anticoagulants. After the blood samples were centrifuged at 1400g for 10 min, the serum was fractionated and pooled. Human IgG was purified using the Melon Gel IgG Spin Purification kit as described above, and the purified IgG was stored at -70 °C for subsequent use in culture experiments.

Both purified IgGs were sterilized using 0.20-micron filters (Corning, Germany), and their IgG concentrations were determined with the Coomassie Protein Assay Reagent (Pierce, USA) according to the manufacturer’s instructions.

Murine immunization

Female WT mice were immunized subcutaneously with 6 mg of Alum (FURP, Sao Paulo) only or supplemented with 1500 µg of OVA (EndoFit™—endotoxin levels Sao Paulo School of Medicine (CEP-FMUSP: 122/14—Sao Paulo, SP, Brazil). The Human Ethics Committee at the School of Medicine at the University of São Paulo approved this study. Informed consent was obtained from all of the volunteers (CAAE: 15507613.4.0000.0060). All authors consented to the publication.

Funding

This study was funded through a grant from the Laboratory of Medical Investigation-56, Medical School, University of Sao Paulo, Sao Paulo, Brazil (LIM-56 HC-FMUSP), Grants #2015/17256-3, #2013/22820-0 and #2010/09004-0 from the São Paulo Research Foundation (FAPESP), and Grant #115603/2015-8 from The National Council for Scientific and Technological Development (CNPq).

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/ ), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/ ) applies to the data made available in this article, unless otherwise stated.

Additional files

13223_2017_195_MOESM1_ESM.pdf Additional file 1: Figure S1. Illustrative dot plots of gating strategy to identify B cells that produces IL-10 and that express B10 phenotype on offspring spleen. Each sample was acquired in the singlet cells gate (determined by FSC-A/FSC-H parameters), panels represent gate strategy then in the lymphocytes gate (determined by their relative size/granularity), and then gated as CD19+ cells (B cells), IL-10 (IL-10+B cells) and CD1dhigh cells (B10 cells).

Authors’ Affiliations

Division of Pathology, Medical School, University of Sao Paulo

References

Fusaro A, Maciel M, Victor J, Oliveira C, Duarte A, Sato M. Influence of maternal murine immunization with Dermatophagoides pteronyssinus extract on the type I hypersensitivity response in offspring. Int Arch Allergy Immunol. 2002;127:208–16. View Article PubMed Google Scholar

Fusaro AE, Brito CA, Victor JR, Rigato PO, Goldoni AL, Duarte AJS, et al. Maternal-fetal interaction: preconception immunization in mice prevents neonatal sensitization induced by allergen exposure during pregnancy and breastfeeding. Immunology. 2007;122:107–15. View Article PubMed PubMed Central Google Scholar

Fusaro AE, de Brito CA, Taniguchi EF, Muniz BP, Victor JR, Orii NM, et al. Balance between early life tolerance and sensitization in allergy: dependence on the timing and intensity of prenatal and postnatal allergen exposure of the mother. Immunology. 2009;128:e541–50. View Article PubMed PubMed Central Google Scholar

Rigato PO, Fusaro AE, Victor JR, Sato MN. Maternal immunization to modulate the development of allergic response and pathogen infections. Immunotherapy. 2009;1:141–56. View Article PubMed Google Scholar

Victor J, Fusaro A, Duarte A, Sato M. Preconception maternal immunization to dust mite inhibits the type I hypersensitivity response of offspring. J Allergy Clin Immunol. 2003;111:269–77. View Article PubMed Google Scholar

Muniz BP, Victor JR, Oliveira LdM, de Lima Lira AA, Perini A, Olivo CR, et al. Tolerogenic microenvironment in neonatal period induced by maternal immunization with ovalbumin. Immunobiology. 2014;219:377–84. View Article PubMed Google Scholar

Victor JR, Muniz BP, Fusaro AE, de Brito CA, Taniguchi EF, Duarte AJS, et al. Maternal immunization with ovalbumin prevents neonatal allergy development and up-regulates inhibitory receptor Fc gamma RIIB expression on B cells. BMC Immunol. 2010;11:11. View Article PubMed PubMed Central Google Scholar

Seeger M, Thierse HJ, Lange H, Shaw L, Hansen H, Lemke H. Antigen-independent suppression of the IgE immune response to bee venom phospholipase A2 by maternally derived monoclonal IgG antibodies. Eur J Immunol. 1998;28:2124–30. View Article PubMed Google Scholar

Rabinovitch N, Gelfand EW. Expression of functional activating and inhibitory Fc gamma receptors on human B cells. Int Arch Allergy Immunol. 2004;133:285–94. View Article PubMed Google Scholar

Noh G, Lee JH. Regulatory B cells and allergic diseases. Allergy Asthma Immunol Res. 2011;3:168–77. View Article PubMed PubMed Central Google Scholar

Amu S, Saunders SP, Kronenberg M, Mangan NE, Atzberger A, Fallon PG. Regulatory B cells prevent and reverse allergic airway inflammation via FoxP3-positive T regulatory cells in a murine model. J Allergy Clin Immunol. 2010;125(1114–24):e8. Google Scholar

Heinzerling L, Mari A, Bergmann KC, Bresciani M, Burbach G, Darsow U, et al. The skin prick test—European standards. Clin Transl Allergy. 2013;3:3. View Article PubMed PubMed Central Google Scholar

Lira AA, de Oliveira MG, de Oliveira LM, Duarte AJ, Sato MN, Victor JR. Maternal immunization with ovalbumin or Dermatophagoides pteronyssinus has opposing effects on Fc?RIIb expression on offspring B cells. Allergy Asthma Clin Immunol. 2014;10:47. View Article PubMed PubMed Central Google Scholar

van Rijt LS, Kuipers H, Vos N, Hijdra D, Hoogsteden HC, Lambrecht BN. A rapid flow cytometric method for determining the cellular composition of bronchoalveolar lavage fluid cells in mouse models of asthma. J Immunol Methods. 2004;288:111–21. View Article PubMed Google Scholar

Verhasselt V, Milcent V, Cazareth J, Kanda A, Fleury S, Dombrowicz D, et al. Breast milk-mediated transfer of an antigen induces tolerance and protection from allergic asthma. Nat Med. 2008;14:170–5. View Article PubMed Google Scholar

Li J, Shen C, Liu Y, Li Y, Sun L, Jiao L, et al. Impaired function of CD5 + CD19 + CD1dhi B10 cells on IgE secretion in an atopic dermatitis-like mouse model. PLoS ONE. 2015;10:e0132173. View Article PubMed PubMed Central Google Scholar

Lira AADL, Oliveira MGD, Oliveira LMD, Duarte AJDS, Sato MN, Victor JR. Opposite effect on offspring Fc gamma RIIb B cells expression on dependency of maternal immunisation with OVA or Dermatophagoides pteronyssinus: different mechanisms for different allergens? Allergy. 2014;69:344. Google Scholar

Victor JR. Influence of maternal immunization with allergens on the thymic maturation of lymphocytes with regulatory potential in children: a broad field for further exploration. J Immunol Res. 2014;2014:780386. View Article PubMed PubMed Central Google Scholar

Bento-de-Souza L, Victor JR, Bento-de-Souza LC, Arrais-Santos M, Rangel-Santos AC, Pereira-Costa É, et al. Constitutive expression of genes encoding notch receptors and ligands in developing lymphocytes, nTreg cells and dendritic cells in the human thymus. Results Immunol. 2016;6:15–20. View Article PubMed PubMed Central Google Scholar

Stadler BM, Zürcher AW, Miescher S, Kricek F, Vogel M. Mimotope and anti-idiotypic vaccines to induce an anti-IgE response. Int Arch Allergy Immunol. 1999;118:119–21. View Article PubMed Google Scholar

Jin G, Hamaguchi Y, Matsushita T, Hasegawa M, Le Huu D, Ishiura N, et al. B-cell linker protein expression contributes to controlling allergic and autoimmune diseases by mediating IL-10 production in regulatory B cells. J Allergy Clin Immunol. 2013;131:1674–82. View Article PubMed Google Scholar

Fink K, Zellweger R, Weber J, Manjarrez-Orduno N, Holdener M, Senn BM, et al. Long-term maternal imprinting of the specific B cell repertoire by maternal antibodies. Eur J Immunol. 2008;38:90–101. View Article PubMed Google Scholar

Victor JR. Allergen-specific IgG as a mediator of allergy inhibition: lessons from mother to child. Hum Vaccin Immunother. 2016. doi: 10.1080/21645515.2016.1244592 . PubMed PubMed Central Google Scholar

Flicker S, Marth K, Kofler H, Valenta R. Placental transfer of allergen-specific IgG but not IgE from a specific immunotherapy-treated mother. J Allergy Clin Immunol. 2009;124:1358–60. View Article PubMed Google Scholar

Glovsky MM, Ghekiere L, Rejzek E. Effect of maternal immunotherapy on immediate skin test reactivity, specific rye I IgG and IgE antibody, and total IgE of the children. Ann Allergy. 1991;67:21–4. PubMed Google Scholar