Modulation of Human Colostrum Phagocyte Activity by the Glycine-Adsorbed Polyethylene Glycol Microspheres

Colostrum is a secretion that contains immunologically active components, including immunocompetent cells and glycine, which has anti-inflammatory, immunomodulatory, and cytoprotective effects. The aim of this study was to evaluate the adsorption of glycine onto polyethylene glycol (PEG) microspheres and to verify the immunomodulatory effect of this nanomaterial on human colostrum phagocytes. The PEG microspheres were evaluated by fluorescence microscopy. The effects of PEG microspheres with adsorbed glycine on viability, superoxide release, phagocytosis, microbicidal activity, and intracellular calcium release of mononuclear (MN) and polymorphonuclear (PMN) colostrum phagocytes were determined. Fluorescence microscopy analyses revealed that glycine was able to be adsorbed to the PEG microspheres. The PMN phagocytes exposed to glycine-PEG microspheres showed the highest superoxide levels. The phagocytes (both MN and PMN) displayed increased microbicidal activity and intracellular calcium release in the presence of PEG microspheres with adsorbed glycine. These data suggest that the adsorption of PEG microspheres with adsorbed glycine was able to stimulate the colostrum phagocytes. This material may represent a possible alternative therapy for future clinical applications on patients with gastrointestinal infections

Imunologia das interações materno-fetais no diabete e na hiperglicemia gestacional leve

In diabetes there is controversy regarding the balance between proinflammatory and anti-inflammatory cytokines, but there is lacking in the literature studies on the cytokine profile in maternal blood and colostrum of diabetic mothers. This study evaluated the subsets of cells and cytokine profile Th1, Th2 e Th17 of maternal blood, cord blood and colostrum of diabetic mothers and verify the modulation of cytokines on the functional activity of phagocytes. Were evaluated 124 pregnant women that were divided according to their glycemic status: non-diabetic (ND, ND=45), mild gestational hyperglycemia (HGL, N=30), gestational diabetes mellitus (GDM, N=15) and diabetes mellitus type 2 (DM2, N=34). We observed that the percentages of CD3+ CD45RA+ cells was lower in cord blood group DM2. The CD4+ cells were lower in cord blood groups with HGL and DM2. Maternal blood group DM2 exhibited reduced subset of CD45RA+ cells. Subsets of CD45RO+ cells increased in the colostrum of mothers with DM2. The higher percentage of CD4+ cells were observed in maternal blood and colostrum of mothers with DM2. The concentration of cytokines IL2, IL4 and IFN-γ were similar in all groups. The concentration of IL-6 was higher in colostrum when compared to concentrations in maternal and cord blood. Higher concentrations of IL-17 were found in the maternal blood DM2. In relation of the functional activity of phagocytes was observed that the mononuclear cells of colostrum and blood treated or not with cytokines (IFN-γ and TGF-β), independent of blood glucose levels, exhibited similar phagocytic activity for enteropathogenic Escherichia coli (EPEC). In DM2 group, irrespective of the presence of cytokines phagocytes maternal blood showed lower release of intracellular calcium. Phagocytes of the cord blood of the non-diabetic group when treated by TGF-β, showed an increase in the release of intracellular calcium. The cells of colostrum and maternal blood in the ...No diabetes há controvérsias em relação ao equilíbrio entre citocinas pró-inflamatórias e anti-inflamatórias, sendo carente na literatura estudos sobre o perfil de citocinas no sangue materno e no colostro de mães diabéticas. Assim, este estudo avaliou a imunofenotipagem de leucócitos e os perfis de respostas Th1, Th2 e Th17 no sangue materno, no sangue de cordão umbilical e no colostro de mães diabéticas e verificar a modulação de citocinas sobre a atividade funcional de fagócitos. Foram avaliadas 124 gestantes, assim distribuídas: não-diabético (ND; N=45), hiperglicemia gestacional leve (HGL; N=30), diabetes mellitus gestacional (DMG; N=15) e diabetes mellitus tipo 2 (DM2; N = 34). Observou-se que as porcentagens de células CD3+CD45RA+ foram menores no sangue do cordão do grupo DM2. As células TCD4+ foram menores no sangue do cordão dos grupos com HGL e DM2. O sangue materno do grupo de DM2 apresentou diminuição das subpopulações de células CD45RA+. As subpopulações de células CD45RO+ aumentaram no colostro de mães portadoras de DM2. O maior percentual de células TCD4+ foi observado no sangue materno e colostro de mães com DM2. Com relação a concentração de citocinas, observou que as concentrações de IL2, IL4 e IFN-γ foram similares em todos os grupos estudados. A concentração de IL6 foi superior no colostro quando comparado às concentrações encontradas no sangue materno e no sangue do cordão. As maiores concentrações de IL-17 foram observadas no sangue materno do grupo DM2. Na avaliação da atividade funcional de fagócitos observou-se que as células mononucleares do colostro e do sangue materno, tratados ou não com as citocinas (IFN-γ e TGF-β) e independente dos níveis glicêmico, exibiram índices fagocíticos similares para Escherichia coli enteropatogênica (EPEC). No grupo DM2, independente da presença de citocinas os fagócitos do sangue materno apresentaram menor liberação de ...Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Melatonin Nanoparticles Adsorbed to Polyethylene Glycol Microspheres as Activators of Human Colostrum Macrophages

The effectiveness of hormones associated with polymeric matrices has amplified the possibility of obtaining new drugs to activate the immune system. Melatonin has been reported as an important immunomodulatory agent that can improve many cell activation processes. It is possible that the association of melatonin with polymers could influence its effects on cellular function. Thus, this study verified the adsorption of the hormone melatonin to polyethylene glycol (PEG) microspheres and analyzed its ability to modulate the functional activity of human colostrum phagocytes. Fluorescence microscopy and flow cytometry analyses revealed that melatonin was able to adsorb to the PEG microspheres. This system increased the release of superoxide and intracellular calcium. There was an increase of phagocytic and microbicidal activity by colostrum phagocytes when in the presence of melatonin adsorbed to PEG microspheres. The modified delivery of melatonin adsorbed to PEG microspheres may be an additional mechanism for its microbicidal activity and represents an important potential treatment for gastrointestinal infections of newborns

The Role of Cytokines in the Functional Activity of Phagocytes in Blood and Colostrum of Diabetic Mothers

Immune response changes induced by diabetes are a risk factor for infections during pregnancy and may modify the development of the newborn's immune system. The present study analyzed colostrum and maternal and cord blood of diabetic women to determine (1) the levels of the cytokines IFN-gamma and TGF-beta and (2) phagocytic activity after incubation with cytokines. Methods. Colostrum and maternal and cord blood samples were classified into normoglycemic (N = 20) and diabetic (N = 19) groups. Cytokine levels, superoxide release, rate of phagocytosis, bactericidal activity, and intracellular Ca2+ release by phagocytes were analyzed in the samples. Irrespective of glycemic status, IFN-gamma and TGF-beta levels were not changed in colostrum and maternal and cord blood. In maternal blood and colostrum, superoxide release by cytokine-stimulated phagocytes was similar between the groups. Compared to spontaneous release, superoxide release was stimulated by IFN-gamma and TGF-beta in normoglycemic and diabetic groups. In the diabetic group, cord blood phagocytes incubated with IFN-gamma exhibited higher phagocytic activity in response to EPEC, and maternal blood exhibited lower microbicidal activity. These data suggest that diabetes interferes in maternal immunological parameters and that IFN-gamma and TGF-beta modulate the functional activity of phagocytes in the colostrum, maternal blood, and cord blood of pregnant diabetic women.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

The effect of IFN-γ and TGF-β in the functional activity of mononuclear cells in the presence of Entamoeba histolytica

Abstract Background Entamoeba histolytica (E. histolytica) causes amoebiasis, which is a disease with significant morbidity and mortality. Phagocytic cells and cytokines appear to be important in amoebiasis, but very little is known about the influence of these cells and cytokines in protozoan infections. The aim of this study was to analyse the supernatant of cultures of mononuclear (MN) cells with E. histolytica to determine: 1) the levels of the cytokines IFN-γ and TGF-β, and 2) the amoebicidal activity of MN cells after incubation with cytokines. Methods Blood samples were collected from 30 volunteer donors. The cytokine concentrations in MN cells culture supernatants, superoxide release, leukophagocytosis, amoebicide activity, intracellular calcium release and apoptosis were analysed. Results The IFN-γ concentrations were 6.22 ± 0.36 and TGF-β concentrations were 17.01 ± 2.21 in cells–trophozoite culture supernatants. MN cells, independently of cytokines, in the presence of amoeba increase the superoxide release. In the absence of cytokines, the ingestion of MN cells by amoebae was higher. In the presence of IFN- γ or TGF- β, a lower ingestion of MN cells was observed by amoebae. MN cells treated with cytokines exhibited higher amoebicide and apoptosis indexes. The incubation of cytokines increased the intracellular calcium release by MN cells. Conclusions These results suggest that cytokines play a beneficial role for the host by activating MN cells against E. histolytica. The increased death of amoebae during the leukophagocytosis suggests that both cytokines (IFN-γ and TGF-β) can modulate the functional activity of MN cells and that these cytokines probably are important in the control of amoebic infections

Modulation of Human Colostrum Phagocyte Activity by the Glycine-Adsorbed Polyethylene Glycol Microspheres

Colostrum is a secretion that contains immunologically active components, including immunocompetent cells and glycine, which has anti-inflammatory, immunomodulatory, and cytoprotective effects. The aim of this study was to evaluate the adsorption of glycine onto polyethylene glycol (PEG) microspheres and to verify the immunomodulatory effect of this nanomaterial on human colostrum phagocytes. The PEG microspheres were evaluated by fluorescence microscopy. The effects of PEG microspheres with adsorbed glycine on viability, superoxide release, phagocytosis, microbicidal activity, and intracellular calcium release of mononuclear (MN) and polymorphonuclear (PMN) colostrum phagocytes were determined. Fluorescence microscopy analyses revealed that glycine was able to be adsorbed to the PEG microspheres. The PMN phagocytes exposed to glycine-PEG microspheres showed the highest superoxide levels. The phagocytes (both MN and PMN) displayed increased microbicidal activity and intracellular calcium release in the presence of PEG microspheres with adsorbed glycine. These data suggest that the adsorption of PEG microspheres with adsorbed glycine was able to stimulate the colostrum phagocytes. This material may represent a possible alternative therapy for future clinical applications on patients with gastrointestinal infections

Secretory IgA-Fc alpha receptor interaction modulating phagocytosis and microbicidal activity by phagocytes in human colostrum of diabetics

Franc, a EL, Morceli G, Fagundes DLG, Rudge MVC, Calderon I de MP, Honorio-Franca AC. Secretory IgA-Fc alpha receptor interaction modulating phagocytosis and microbicidal activity by phagocytes in human colostrum of diabetics. APMIS 2011; 119: 710-19.The effects of secretory immunoglobulin A (SIgA) interaction with its specific Fc alpha receptors on colostral phagocytes needs further investigation, especially with respect to diabetic women. Accordingly, we studied the colostrum of hyperglycemic women to assess SIgA interactions with Fc alpha receptors of macrophages as well as the functional activity of these cells. The women were divided for colostrum sampling according to their glycemic status: normoglycemia (N = 51), mild hyperglycemia (N = 23), and diabetes (N = 25) groups. We determined the Fc alpha R expression, the IgA on the surface and the surface-bound IgA in colostrum macrophages. We also evaluated the superoxide release and bactericidal killing of these cells. Colostral phagocytes expressed Fc alpha R, contained IgA on the surface and are able to bind to purified SIgA. The bactericidal activity of colostral phagocytes from the hyperglycemic women was similar to that of normoglycemic only when SIgA was used as opsonin. Addition of a MoAb anti-human Fc alpha receptor resulted in a significant decrease of superoxide release and bacterial killing by macrophages when bacteria were opsonized with purified SIgA, suggesting an interaction between SIgA and Fc alpha R. The stimulatory effects of SIgA on the functional activity of phagocytes therefore protect infants, especially of diabetic women, against intestinal infections.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Changes in T-cell phenotype and cytokines profile in maternal blood, cord blood and colostrum of diabetic mothers

The present study evaluated the cells and cytokine of maternal blood, cord blood and colostrum of diabetic mothers. The women evaluated were divided according to their body mass index (BMI) and glycemic status into non-diabetic (ND - N = 15), mild gestational hyperglycemic (MGH - N = 15), diabetes mellitus gestational (DMG - N = 13) and type-2 diabetes mellitus (DM2 - N = 15) groups. The subsets of cells and cytokine profile were determined by flow cytometry. Maternal blood from MGH group had increase percentage of CD3(+)T cells, and DM-2 group had decrease percentage of CD4(+) T cells. The cord blood from hyperglycemic groups showed lower percentage of CD3(+) T cells expressing CD45RO(+) and higher of CD4(+) T cells and CD4(+) T cells expressing CD45RA(+). In the colostrum, the CD4(+) T cells and CD4(+) T cells expressed CD45RA(+) increase in hyperglycemic groups. The DM2 group exhibited higher IL17 levels in maternal blood. IFN-γ was lower in cord blood from MGH and DMG groups with overweight/obese. Irrespective of the glycemic status, IL6 was higher in colostrum. The results obtained suggest that maternal hyperglycemia modifies the phenotypes of T cells and cytokines profile in maternal, cord blood and colostrum