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1. Exploring the Immunomodulatory Potential of Human Milk: Aryl Hydrocarbon Receptor Activation and Its Impact on Neonatal Gut Health

Author

Naomi V. Wieser, Mohammed Ghiboub, Caroline Verseijden, Johannes B. van Goudoever, Anne Schoonderwoerd, Tim G. J. de Meij, Hendrik J. Niemarkt, Mark Davids, Antoine Lefèvre, Patrick Emond, Joep P. M. Derikx, Wouter J. de Jonge, and Bruno Sovran

Subjects
prematurity, aryl hydrocarbon receptor, barrier integrity, Nutrition. Foods and food supply, TX341-641
Abstract

Several metabolites of the essential amino acid tryptophan have emerged as key players in gut homeostasis through different cellular pathways, particularly through metabolites which can activate the aryl hydrocarbon receptor (AHR). This study aimed to map the metabolism of tryptophan in early life and investigate the effects of specific metabolites on epithelial cells and barrier integrity. Twenty-one tryptophan metabolites were measured in the feces of full-term and preterm neonates as well as in human milk and formula. The ability of specific AHR metabolites to regulate cytokine-induced IL8 expression and maintain barrier integrity was assessed in Caco2 cells and human fetal organoids (HFOs). Overall, higher concentrations of tryptophan metabolites were measured in the feces of full-term neonates compared to those of preterm ones. Within AHR metabolites, indole-3-lactic acid (ILA) was significantly higher in the feces of full-term neonates. Human milk contained different levels of several tryptophan metabolites compared to formula. Particularly, within the AHR metabolites, indole-3-sulfate (I3S) and indole-3-acetic acid (IAA) were significantly higher compared to formula. Fecal-derived ILA and milk-derived IAA were capable of reducing TNFα-induced IL8 expression in Caco2 cells and HFOs in an AHR-dependent manner. Furthermore, fecal-derived ILA and milk-derived IAA significantly reduced TNFα-induced barrier disruption in HFOs.

Published
2024
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2. Human Milk from Previously COVID-19-Infected Mothers: The Effect of Pasteurization on Specific Antibodies and Neutralization Capacity

Author

Britt J. van Keulen, Michelle Romijn, Albert Bondt, Kelly A. Dingess, Eva Kontopodi, Karlijn van der Straten, Maurits A. den Boer, Judith A. Burger, Meliawati Poniman, Berend J. Bosch, Philip J. M. Brouwer, Christianne J. M. de Groot, Max Hoek, Wentao Li, Dasja Pajkrt, Rogier W. Sanders, Anne Schoonderwoerd, Sem Tamara, Rian A. H. Timmermans, Gestur Vidarsson, Koert J. Stittelaar, Theo T. Rispens, Kasper A. Hettinga, Marit J. van Gils, Albert J. R. Heck, and Johannes B. van Goudoever

Subjects
immunoglobulins, pasteurization, COVID-19, breastfeeding, Nutrition. Foods and food supply, TX341-641
Abstract

Background: Since the outbreak of coronavirus disease 2019 (COVID-19), many put their hopes in the rapid availability of effective immunizations. Human milk, containing antibodies against syndrome coronavirus 2 (SARS-CoV-2), may serve as means of protection through passive immunization. We aimed to determine the presence and pseudovirus neutralization capacity of SARS-CoV-2 specific IgA in human milk of mothers who recovered from COVID-19, and the effect of pasteurization on these antibodies. Methods: This prospective case control study included lactating mothers, recovered from (suspected) COVID-19 and healthy controls. Human milk and serum samples were collected. To assess the presence of SARS-CoV-2 antibodies we used multiple complementary assays, namely ELISA with the SARS-CoV-2 spike protein (specific for IgA and IgG), receptor binding domain (RBD) and nucleocapsid (N) protein for IgG in serum, and bridging ELISA with the SARS-CoV-2 RBD and N protein for specific Ig (IgG, IgM and IgA in human milk and serum). To assess the effect of pasteurization, human milk was exposed to Holder (HoP) and High Pressure Pasteurization (HPP). Results: Human milk contained abundant SARS-CoV-2 antibodies in 83% of the proven cases and in 67% of the suspected cases. Unpasteurized milk with and without these antibodies was found to be capable of neutralizing a pseudovirus of SARS-CoV-2 in (97% and 85% of the samples respectively). After pasteurization, total IgA antibody levels were affected by HoP, while SARS-CoV-2 specific antibody levels were affected by HPP. Pseudovirus neutralizing capacity of the human milk samples was only retained with the HPP approach. No correlation was observed between milk antibody levels and neutralization capacity. Conclusions: Human milk from recovered COVID-19-infected mothers contains SARS-CoV-2 specific antibodies which maintained neutralization capacity after HPP. All together this may represent a safe and effective immunization strategy after HPP.

Published
2021
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3. Breastmilk; a source of SARS-CoV-2 specific IgA antibodies

Author

Britt J. van Keulen, Michelle Romijn, Albert Bondt, Kelly A. Dingess, Eva Kontopodi, Karlijn van der Straten, Maurits A. den Boer, Berend J. Bosch, Philip J.M. Brouwer, Christianne J.M. de Groot, Max Hoek, Wentao Li, Dasja Pajkrt, Rogier W. Sanders, Anne Schoonderwoerd, Sem Tamara, Rian A.H. Timmermans, Gestur Vidarsson, Koert J. Stittelaar, Theo T. Rispens, Kasper A. Hettinga, Marit J. van Gils, Albert J.R. Heck, and Johannes B. van Goudoever

Subjects
viruses, skin and connective tissue diseases
Abstract

BackgroundSince the outbreak of COVID-19, many put their hopes in the rapid development of effective immunizations. For now patient isolation, physical distancing and good hygiene are the sole measures for prevention. Processed breast milk with antibodies against SaRS-CoV-2 may serve as additional protection. We aimed to determine the presence and neutralization capacity of antibodies against SaRS-CoV-2 in breastmilk of mothers who have recovered from COVID-19.MethodsThis prospective case control study included lactating mothers, recovered from (suspected) COVID-19 and healthy controls. Serum and breastmilk was collected. To assess the presence of antibodies in breastmilk and serum, we used multiple complementary assays, namely ELISA with the SARS-CoV-2 spike protein, SARS-CoV-2 receptor binding domain (RBD) and with the SARS-CoV-2 nucleocapsid (N) protein for IgG and bridging ELISA with the SARS-CoV-2 RBD and N protein for total Ig. To assess the effect of pasteurization breastmilk was exposed to Holder Pasteurization and High Pressure Pasteurization.ResultsBreastmilk contained antibodies against SARS-CoV-2 using any of the assays in 24 out of 29 (83%) proven cases, in six out of nine (67%) suspected cases and in none of the 13 controls.In vitroneutralization of SARS-CoV-2 clinical isolate virus strain was successful in a subset of serum (13%) and milk samples (26%). Although after pasteurization of the milk SARS-CoV-2 antibodies were detected with both methods of pasteurization, virus neutralizing capacity of those antibodies was only retained with the HPP approach.ConclusionBreastmilk of mothers who recovered from COVID-19 contains significant amounts of IgA against SARS-CoV-2, both before and after pasteurization.Key PointsQuestionDoes breastmilk of mothers who have recovered from coronavirus disease 2019 (COVID-19) contain antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)?FindingsWe provide multiple lines of evidence on the presence of a variety of antibodies against SARS-CoV-2, with no such antibodies present in the controls. These antibodies are capable of neutralizing a clinical isolate of SARS-CoV-2in vitro. We furthermore show that high pressure pasteurization hardly affects antibody levels and efficacy.MeaningBreastmilk, obtained from mothers who have recovered from COVID-19, may serve as a safe and widely applicable preventive strategy for vulnerable high risk populations

Published
2020

4. Breastmilk: A Source of SARS-CoV-2 Specific IgA Antibodies

Author

Albert J. R. Heck, Eva Kontopodi, Kelly A Dingess, Koert J. Stittelaar, Gestur Vidarsson, Max Hoek, Sem Tamara, Anne Schoonderwoerd, Theo Rispens, Christianne J.M. de Groot, Berend J. Bosch, Roger W Sanders, Michelle Romijn, Marit J. van Gils, Maurits A. den Boer, R.A.H. Timmermans, Karlijn van der Straten, Kasper Hettinga, Dasja Pajkrt, Wentao Li, Albert Bondt, Philip J. M. Brouwer, Johannes B. van Goudoever, and Britt J. van Keulen

Subjects
biology, business.industry, viruses, Case-control study, Outbreak, Pasteurization, Breast milk, Virology, Virus, Neutralization, law.invention, medicine.anatomical_structure, law, Lactation, biology.protein, Medicine, Antibody, skin and connective tissue diseases, business
Abstract

Background Since the outbreak of COVID-19, many put their hopes in the rapid development of effective immunizations. For now patient isolation, physical distancing and good hygiene are the sole measures for prevention. Processed breast milk with antibodies against SaRS-CoV-2 may serve as additional protection. We aimed to determine the presence and neutralization capacity of antibodies against SaRS-CoV-2 in breastmilk of mothers who have recovered from COVID-19. Methods This prospective case control study included lactating mothers, recovered from (suspected) COVID-19 and healthy controls. Serum and breastmilk was collected. To assess the presence of antibodies in breastmilk and serum, we used multiple complementary assays, namely ELISA with the SARS-CoV-2 spike protein, SARS-CoV-2 receptor binding domain (RBD) and with the SARS-CoV-2 nucleocapsid (N) protein for IgG and bridging ELISA with the SARS-CoV-2 RBD and N protein for total Ig. To assess the effect of pasteurization breastmilk was exposed to Holder Pasteurization and High Pressure Pasteurization. Results Breastmilk contained antibodies against SARS-CoV-2 using any of the assays in 24 out of 29 (83%) proven cases, in six out of nine (67%) suspected cases and in none of the 13 controls. In vitro neutralization of SARS-CoV-2 clinical isolate virus strain was successful in a subset of serum (13%) and milk samples (26%). Although after pasteurization of the milk SARS-CoV-2 antibodies were detected with both methods of pasteurization, virus neutralizing capacity of those antibodies was only retained with the HPP approach. Conclusion Breastmilk of mothers who recovered from COVID-19 contains significant amounts of IgA against SARS-CoV-2, both before and after pasteurization. Key Points Question Does breastmilk of mothers who have recovered from coronavirus disease 2019 (COVID-19) contain antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)? Findings We provide multiple lines of evidence on the presence of a variety of antibodies against SARS-CoV-2, with no such antibodies present in the controls. These antibodies are capable of neutralizing a clinical isolate of SARS-CoV-2 in vitro. We furthermore show that high pressure pasteurization hardly affects antibody levels and efficacy. Meaning Breastmilk, obtained from mothers who have recovered from COVID-19, may serve as a safe and widely applicable preventive strategy for vulnerable high risk populations

Published
2020

5. Holder-Pasteurized Human Donor Milk: How Long Can It Be Preserved?

Author

Marita de Waard, Elise Mank, Karin van Dijk, Johannes B. van Goudoever, Anne Schoonderwoerd, Pediatric surgery, Medical Microbiology and Infection Prevention, VU University medical center, ACS - Diabetes & metabolism, AGEM - Endocrinology, metabolism and nutrition, Amsterdam Reproduction & Development (AR&D), Graduate School, Neonatology, and Amsterdam Gastroenterology Endocrinology Metabolism

Subjects
Adult, 0301 basic medicine, Time Factors, Pasteurization, Safety standards, complex mixtures, law.invention, 03 medical and health sciences, fluids and secretions, 0302 clinical medicine, law, 030225 pediatrics, parasitic diseases, Humans, Medicine, Prospective Studies, Food science, Cryopreservation, 030109 nutrition & dietetics, Milk, Human, business.industry, Gastroenterology, food and beverages, humanities, Milk Banks, Pediatrics, Perinatology and Child Health, Female, business, Follow-Up Studies
Abstract

Objective: When own mother's milk falls short, pasteurized human donor milk is recommended as alternative feeding for preterm infants. Donor milk has to meet the highest safety standards, but its processing and storage is expensive. The recommended storage time of pasteurized donor milk is 3 months. The objective of the present study was to determine whether the frozen storage time of pasteurized donor milk can be extended beyond 3 months without compromising its safety and quality. Methods: For this prospective observational study breast milk samples of 34 unique women, collected between November 2014 and June 2015, were provided by the Dutch Human Milk Bank. Samples were Holder pasteurized within 3 months after expression and stored at -20°C. Analysis of both bacterial growth (by inoculation of milk on a blood and a cysteine-, lactose-, and electrolyte-deficient agar) and fat, crude protein, carbohydrate and energy content of milk (analyzed by infrared spectroscopy) was done monthly during the first 6 months and every 2 months thereafter, up to 1 year postpasteurization. Results: Thirty of 306 (9.8%) follow-up samples showed bacterial growth when cultured. None of the samples showed sequential contamination with the same strain up to 8 months of frozen storage. No significant decreases in macronutrients and energy content were observed over 8 months. Conclusion: Pasteurized human donor milk can be stored safely for 8 months at -20°C, without compromising its macronutrient and energy content. This longer storage time will reduce disposal of expired donor milk and subsequently reduce costs.

Published
2018

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