Gestation and breastfeeding in schistosomotic mice differentially alters the expression of histone deacetylases (HDACs) in adult offspring

BACKGROUND Breastfeeding or gestation in schistosomotic mothers can cause long-term alterations in the immune response of offspring. OBJECTIVES Evaluate the expression of histone deacetylases (HDACs) (all classes), the production of cytokines by T and B lymphocytes and macrophages, and the frequency of CD4+CD25+FoxP3+-cells in adult offspring born and/or suckled by schistosomotic mothers. METHODS We harvested splenocytes from offspring born to (BIM), suckled by (SIM), or born to/suckled by (BSIM) schistosomotic mothers and animals from noninfected mothers (Control) at seven-weeks old and cultured them with/without Concanavalin A. HDAC expression was evaluated by real-time quantitative polymerase chain reaction (qPCR), and cytokines and membrane markers were evaluated by fluorescence-activated cell sorting (FACS). FINDINGS Compared to Control, BIM mice showed increased expression of HDAC9 and frequency of CD4+IL-10+-cells. The SIM group had increased expression of HDAC1, HDAC2, HDAC6, HDAC7, HDAC10, Sirt2, Sirt5, Sirt6, and Sirt7. The BSIM group only had increased HDAC10 expression. The SIM and BSIM groups exhibited decreased frequencies of CD4+IL-4+-cells and CD4+CD25+FoxP3+-cells, along with a higher frequency of CD14+IL-10+-cells and an increase in CD45R/B220+IL-10+-cells. The BSIM group also showed a high frequency of CD4+IL10+-cells. MAIN CONCLUSIONS Breastfeeding induced the expression of HDACs from various classes involved in reducing inflammatory responses. However, gestation enhanced the expression of a single HDAC and breastfeeding or gestation appears to favour multiple IL-10-dependent pathways, but not cells with a regulatory phenotype.

Maternal schistosomiasis: IL-2, IL-10 and regulatory T lymphocytes to unrelated antigen in adult offspring mice

Abstract INTRODUCTION: We evaluated IL-10, IL-2 and regulatory T cells (Treg), in response to ovalbumin (OA), in offspring from schistosomotic mouse mothers. METHODS: We used animals born (BIM) or suckled (SIM) from infected mothers; and mice born/suckled from infected (BSIM) or non-infected mothers (CONTROL). After OA+adjuvant immunization, spleen cells were cultured, with or without OA, and doubly marked for cytometry. RESULTS: BIM showed fewer CD4+/IL-2+ and more B220+/IL-10+ cells, whereas the SIM group showed increased Treg frequency. BSIM had fewer B220+/IL-10+ and Treg cells. CONCLUSIONS: Separately, gestation or nursing induced immunosuppressive cells in infected mothers, but improved anti-OA immunity when combined.

Gestation and breastfeeding in schistosomotic mothers differently modulate the immune response of adult offspring to postnatal Schistosoma mansoni infection

Schistosoma mansoni antigens in the early life alter homologous and heterologous immunity during postnatal infections. We evaluate the immunity to parasite antigens and ovalbumin (OA) in adult mice born/suckled by schistosomotic mothers. Newborns were divided into: born (BIM), suckled (SIM) or born/suckled (BSIM) in schistosomotic mothers, and animals from noninfected mothers (control). When adults, the mice were infected and compared the hepatic granuloma size and cellularity. Some animals were OA + adjuvant immunised. We evaluated hypersensitivity reactions (HR), antibodies levels (IgG1/IgG2a) anti-soluble egg antigen and anti-soluble worm antigen preparation, and anti-OA, cytokine production, and CD4+FoxP3+T-cells by splenocytes. Compared to control group, BIM mice showed a greater quantity of granulomas and collagen deposition, whereas SIM and BSIM presented smaller granulomas. BSIM group exhibited the lowest levels of anti-parasite antibodies. For anti-OA immunity, immediate HR was suppressed in all groups, with greater intensity in SIM mice accompanied of the remarkable level of basal CD4+FoxP3+T-cells. BIM and SIM groups produced less interleukin (IL)-4 and interferon (IFN)-g. In BSIM, there was higher production of IL-10 and IFN-g, but lower levels of IL-4 and CD4+FoxP3+T-cells. Thus, pregnancy in schistosomotic mothers intensified hepatic fibrosis, whereas breastfeeding diminished granulomas in descendants. Separately, pregnancy and breastfeeding could suppress heterologous immunity; however, when combined, the responses could be partially restored in infected descendants.

Transferência passiva de anticorpos específicos para antígenos de Schistosoma mansoni em camundongos nascidos ou amamentados em mães esquistossomóticas / Passive transfer of Schistosoma mansoni antigens-specific antibodies in mice born or suckled by schistosomotic mothers

Objetivo - Estudos experimentais demonstraram que mães infectadas pelo Schistosoma mansoni modulam a imunidade para antígenos homólogos, dos descendentes adultos, através do contato prévio com anticorpos anti-Schistosoma durante o período pré-natal junto à amamentação. Descendentes adultos de mães esquistossomóticas apresentaram alteração na imunidade para um antígeno heterólogo, Ovalbumina (OA): amamentação induziu maior produção de imunoglobulinas anti-OA, enquanto a gestação levou à supressão destas imunoglobulinas. A fim de esclarecer a participação dos anticorpos anti-Schistosoma maternos na alteração da imunidade dos descendentes adultos, os anticorpos contra antígenos solúveis dos ovos (SEA) e dos vermes (SWAP) em descendentes gerados ou apenas amamentados em mães esquistossomóticas foram dosados. Métodos - Camundongos recém-nascidos foram divididos em: animais nascidos de Mães Infectadas (MI) e amamentados em mães não-infectadas; animais nascidos de mães não-infectadas e Amamentados em mães Infectadas (AI); animais nascidos e amamentados em mães infectadas (MIAI) ou não-infectadas (Controle). Os animais foram sangrados 21, 45, 60 e 77 dias, após nascimento e os isótipos IgG1 e IgG2a dosados, no plasma, por ELISA. Resultados - Foi detectado IgG1, mas não IgG2a, principalmente anti-SEA, tanto no grupo MI como nos grupos AI e MIAI. A transferência pela amamentação foi mais efetiva (maiores níveis e manutenção durante a cinética). Conclusões - O isótipo IgG1 anti-SEA presente no grupo MI, bem como no grupo AI, exclui a associação dos anticorpos antiparasita e melhora da imunidade heteróloga dos descendentes amamentados em mães esquistossomótica. Este estudo enfoca o importante papel da amamentação em transferir de forma eficaz anticorpos anti-SEA para indivíduos de área endêmica para esquistossomose.

Objective - Experimental studies have demonstrated that Schistosoma mansoni infected mothers modulate immunity to homologous antigen, in their adult offspring, through prior contact with anti-Schistosoma antibodies during the prenatal period plus breastfeeding. Adult offspring of schistosomotic mothers showed alterations in immunity to a heterologous antigen, ovalbumin (OA): breastfeeding induced higher production of anti-OA immunoglobulin, while the pregnancy led to suppression of this immunoglobulin. In order to study the participation of the maternal anti-Schistosoma antibodies and change in the heterologous immunity in adult offspring, antibodies against soluble egg antigen (SEA) and worms (SWAP) in offspring born or only breastfed by schistosomotic mothers were measured. Methods - Newborn mice were divided into: animals Born from Infected Mothers (BIM) suckled by non-infected mothers; animals from non-infected mothers Suckled by Infected Mothers (SIM); and mice Born and Suckled in Infected Mothers (BSIM) or non-infected (Control) mothers. The animals were bled 21,45, 60, 77 days, after birth, and IgG1 and IgG2a serum isotypes were measured by ELISA. Results - It was detected IgG1, but not IgG2a, mainly anti-SEA in a group BIM and in the groups SIM and BSIM. The transfer by breastfeeding was more effective (higher levels and maintenance during the kinetic). Conclusions - The anti-SEA IgG1 isotype detected in the group BIM, as well as, in the SIM, excludes the association of anti-parasite antibodies and the improvement of heterologous immunity in offspring nursed by schistosomotic mothers. This study highlights the important role of breastfeeding as effective way to transfer anti-SEA antibodies for individuals from an endemic area for schistosomiasis.

Histological and histomorphometric evaluation of the retina of mice infected with Schistosoma mansoni in its hepatosplenic form / Avaliação histológica e histomorfométrica da retina de camundongos infectados pelo Schistosoma mansoni na forma hepatoesplênica

PURPOSE: To evaluate retinal manifestations of Schistosomiasis mansoni in its hepatosplenic form in mice. METHODS: It was performed a study with two groups of mice; one of them was infected with 40 cercariae of Schistosoma mansoni. After 120 days of the infection, the eyes underwent a retinal microscopy study. The histology findings were reported. Histomorphometric analysis was also performed, including: thickness measurement of the retinal layer and the number of the ganglion layer cells. RESULTS: In one case a retinal granuloma was found. The analysis of the other histological sections demonstrated normal architecture of the retina. The mean thickness of the retinal layer between the two groups were similar (41.81±6.09µm versus 38.48±8.58µm - p=0.279); as well as the mean number of the ganglion layer cells (20.93±4.88 versus 20.64±4.10 - p=0.864). Disorganization of the retinal layers was not identified and the histomorphometric analysis revealed no significant difference between the two groups. CONCLUSION: The absence of findings in this study does not exclude that hemodynamic and autoregulation changes associated with hepatosplenic schistosomiasis could be correlated to retinal manifestations. It is necessary that other methods with a high parasite infection should be performed.

OBJETIVO: Avaliar as repercussões da esquistossomose mansônica na forma hepatoesplênica na retina de camundongos. MÉTODOS: Foi realizado estudo com dois grupos de camundongos, sendo um infectado com 40 cercárias do Schistosoma mansoni. Decorridos 120 dias da infecção, os olhos foram submetidos à análise microscópica da retina com descrição dos achados histológicos e realizada análise histomorfométrica com mensuração da espessura de segmento retiniano e do número de células da camada ganglionar. RESULTADOS: Em um caso foi encontrado um granuloma retiniano. Já a análise dos demais cortes histológicos demonstrou uma arquitetura normal da retina. A média da espessura dos segmentos retinianos entre os grupos de camundongos, controle e infectado, foi similar (41,81±6,09µm versus 38,48±8,58µm - p=0,279) assim como a média do número de células da camada ganglionar (20,93±4,88 versus 20,64±4,10 - p=0,864). : A estrutura da retina encontrava-se íntegra e a análise histomorfométrica não revelava diferença significante entre os dois grupos. CONCLUSÃO: A ausência de alterações, neste estudo, não afasta a possibilidade de que desequilíbrios hemodinâmicos e no mecanismo de autoregulação, em portadores da forma hepatoesplênica da esquistossomose, possam acarretar dano retiniano. Demanda, entretanto, que outras metodologias com indução da infecção com uma maior carga parasitária sejam realizadas.