Abstract
Background
Most preterm infants receive antibiotics to prevent serious infections shortly after birth. However, prolonged antibiotic treatment predisposes to gut dysbiosis and late-onset sepsis. Using preterm pigs as model, we hypothesized that neonatal prophylactic antibiotics impair systemic immune development beyond the days of antibiotic treatment.
Methods
Preterm pigs (90% gestation) were fed formula for 9 days, treated with sterile water (CON) or enteral antibiotics from day 1 to 4. On days 5 and 9, blood was collected for haematology, in vitro LPS stimulation, and plasma proteomics.
Results
Antibiotic treatment altered the abundance of 21 and 47 plasma proteins on days 5 and 9, representing 6.6% and 14.8% of the total annotated proteins, respectively. Most antibiotics-induced proteome changes related to complement cascade, neutrophil degranulation, and acute phase responses. Neutrophil and lymphocyte counts were higher in antibiotics-treated pigs on day 5 but did not change from days 5–9, in contrast to increasing cell counts in CON. The antibiotics treatment suppressed TNF-alpha and IL-10 responses to in vitro LPS challenge on day 5, 7 and 9.
Conclusion
Few days of antibiotics treatment following preterm birth alter the plasma proteome and inhibit systemic immune development, even beyond the days of treatment.
Impact
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1.
Neonatal prophylactic antibiotics alter the plasma proteome and suppress systemic immune development in preterm pigs
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2.
The effects of prophylactic antibiotics last beyond the days of treatment.
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3.
Neonatal antibiotics treatment for compromised human newborns may predispose to longer-term risks of impaired immunity and infections.
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Data availability
All datasets generated during and analysed during the current study, including raw data used for all figures, tables and analysis, are available from the corresponding author on reasonable request. Besides, the MS proteomic data are available at the ProteomeXchange Consortium (http://www.proteomexchange.org/) with the data set identifier PXD036276.
References
Reiss, J. D. et al. Perinatal infection, inflammation, preterm birth, and brain injury: A review with proposals for future investigations. Exp. Neurol. 351, 113988 (2022).
Clark, R. H., Bloom, B. T., Spitzer, A. R. & Gerstmann, D. R. Reported medication use in the neonatal intensive care unit: data from a large national data set. Pediatrics 117, 1979 LP–1971987 (2006).
Dishaw, L. J., Carneiro, L. A., Bel, S., Arrieta, M.-C. & Laforest-Lapointe, I. Patterns of early-life gut microbial colonization during human immune development: an ecological. Perspect. Artic. 8, 1 (2017).
Cox, L. M. & Blaser, M. J. Antibiotics in early life and obesity. Nat. Rev. Endocrinol. 11, 182–190 (2015).
Kuppala, V. S., Meinzen-Derr, J., Morrow, A. L. & Schibler, K. R. Prolonged initial empirical antibiotic treatment is associated with adverse outcomes in premature infants. J. Pediatr. 159, 720–725 (2011).
Arrieta, M. C. et al. Early infancy microbial and metabolic alterations affect risk of childhood asthma. Sci. Transl. Med. 7, 307ra152 (2015).
Melville, J. M. & Moss, T. J. M. The immune consequences of preterm birth. Front. Neurosci. 7, 79 (2013).
Bæk, O., Ren, S., Brunse, A., Sangild, P. T. & Nguyen, D. N. Impaired neonatal immunity and infection resistance following fetal growth restriction in preterm pigs. Front. Immunol. 11, 1–13 (2020).
Cotten, C. M. et al. Prolonged duration of initial empirical antibiotic treatment is associated with increased rates of necrotizing enterocolitis and death for extremely low birth weight infants what’s known on this subject. Pediatrics 123, 58–66 (2009).
Walker, W. A. Initial intestinal colonization in the human infant and immune homeostasis key messages. Ann Nutr Metab 63, 8–15 (2013).
Tripathi, N., Cotten, C. M. & Smith, P. B. Antibiotic use and misuse in the neonatal intensive care unit. Clin. Perinatol. 39, 61–68 (2012).
Muk, T., Brunse, A., Henriksen, N. L., Aasmul-Olsen, K. & Nguyen, D. N. Glucose supply and glycolysis inhibition shape the clinical fate of Staphylococcus epidermidis-infected preterm newborns. JCI Insight. 7, e157234 (2022).
Nguyen, D. N. et al. Delayed development of systemic immunity in preterm pigs as a model for preterm infants. Sci. Rep. 6, 36816 (2016).
Jensen, M. L. et al. Antibiotics modulate intestinal immunity and prevent necrotizing enterocolitis in preterm neonatal piglets. Am. J. Physiol. - Gastrointest. Liver Physiol. 306, G59–G71 (2014).
Birck, M. M. et al. Enteral but not parenteral antibiotics enhance gut function and prevent necrotizing enterocolitis in formula-fed newborn preterm pigs. Am. J. Physiol. Gastrointest. Liver Physiol. 310, 323–333 (2016).
Brunse, A. et al. Enteral broad-spectrum antibiotics antagonize the effect of fecal microbiota transplantation in preterm pigs. Gut Microbes 13, 1–16 (2020).
White, A. R. et al. Augmentin (amoxicillin/clavulanate) in the treatment of community-acquired respiratory tract infection: a review of the continuing development of an innovative antimicrobial agent. J. Antimicrob. Chemother. 53 Suppl 1, i3–i20 (2004).
Liu, Y. et al. Pharmacokinetics of neomycin sulfate after intravenous and oral administrations in swine. J. Vet. Pharmacol. Ther. 44, 850–853 (2021).
León, I. R., Schwämmle, V., Jensen, O. N. & Sprenger, R. R. Quantitative assessment of in-solution digestion efficiency identifies optimal protocols for unbiased protein analysis. Mol. Cell. Proteom. 12, 2992–3005 (2013).
Tyanova, S., Temu, T. & Cox, J. The MaxQuant computational platform for mass spectrometry–based shotgun proteomics. Nat. Protoc. 11, 2301–2319 (2016).
Tyanova, S. et al. The Perseus computational platform for comprehensive analysis of (prote)omics data. Nat. Methods 13, 731–740 (2016).
Team, R. C. R: A language and environment for statistical computing. (2013).
Team, Rs. RStudio: integrated development for R. RStudio, Inc., Boston, MA posit.co42, 84 (2015).
Szklarczyk, D. et al. STRING v11: protein-protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets. Nucleic Acids Res. 47, 607–613 (2018).
Masuoka, M. et al. Periostin promotes chronic allergic inflammation in response to Th2 cytokines. J. Clin. Invest. 122, 2590–2600 (2012).
Li, Y. et al. Early use of antibiotics is associated with a lower incidence of necrotizing enterocolitis in preterm, very low birth weight infants: The NEOMUNE-NeoNutriNet Cohort Study. J. Pediatr. 227, 128–134.e2 (2020).
Zwittink, R. D. et al. Association between duration of intravenous antibiotic administration and early-life microbiota development in late-preterm infants. https://doi.org/10.1007/s10096-018-3193-y.
Alexander, V. N., Northrup, V. & Bizzarro, M. J. Antibiotic exposure in the newborn intensive care unit and the risk of necrotizing enterocolitis. J. Pediatr. 159, 392–397 (2011).
Druhan, L. J. et al. Leucine rich α-2 glycoprotein: a novel neutrophil granule protein and modulator of myelopoiesis. https://doi.org/10.1371/journal.pone.0170261 (2017).
Saenko, E. L. et al. Modulatory effects of ceruloplasmin on lymphocytes. neutrophils and monocytes of patients with altered immune status modulatory effects of cerulopiasmin on lymphocytes, neutrophils and monocytes of patients with altered immune status. Immunol. Invest. 23, 99–100 (1994).
Dimberg, J. et al. Expression of the serine protease inhibitor serpinA3 in human colorectal adenocarcinomas. Oncol. Lett. 2, 413–418 (2011).
Li, W. et al. Periostin: its role in asthma and its potential as a diagnostic or therapeutic target. https://doi.org/10.1186/s12931-015-0218-2 (2012).
Conway, S. J. et al. The role of periostin in tissue remodeling across health and disease. Cell. Mol. Life Sci. 71, 1279–1288 (2014).
Liu, A. Y., Zheng, H. & Ouyang, G. Periostin, a multifunctional matricellular protein in inflammatory and tumor microenvironments. Matrix Biol. 37, 150–156 (2014).
Härtel, C. et al. Cytokine responses correlate differentially with age in infancy and early childhood. Clin. Exp. Immunol. 142, 446–453 (2005).
Maródi, L. Innate cellular immune responses in newborns. Clin. Immunol. 118, 137–144 (2006).
Morein, B., Abusugra, I. & Blomqvist, G. Immunity in neonates. Vet. Immunol. Immunopathol. 87, 207–213 (2002).
Cesare, A. D., Meglio, P. D. & Nestle, F. O. A role for Th17 cells in the immunopathogenesis of atopic dermatitis? J. Invest. Dermatol. 128, 2569–2571 (2008).
Grewe, M. et al. A role for Th1 and Th2 cells in the immunopathogenesis of atopic dermatitis. Immunol. Today 19, 359–361 (1998).
De Carli, M. et al. Review human Th1 and Th2 cells: functional properties, regulation of development and role in autoimmunity. Autoimmunity 18, 30–31 (1994).
Dunkelberger, J. R. & Song, W. C. Complement and its role in innate and adaptive immune responses. Cell Res. 20, 34–50 (2010).
Funding
The study was supported by Independent Research Fund Denmark (8022-00188B) and The Danish National Mass Spectrometry Platform for Functional Proteomics (PRO-MS; grant no. 5072-00007B). The Obelske Family Foundation and the Svend Andersen Foundation are acknowledged for grants to the analytical platform enabling parts of this study.
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T.M. contributed to the proteomic data acquirement, data analysis, result interpretation, preparation of the initial draft, and manuscript revision. A.L. and A.S. conducted the proteomics analysis. A.B. contributed to the experimental design, participated in the animal experiment, prepared the proteomics samples and contributed to result interpretation. P.T.S. and T.T. contributed to the experimental design, result interpretation and manuscript preparation. DNN contributed to experimental design and execution, result interpretation and manuscript preparation.
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Muk, T., Leto, A., Brunse, A. et al. Neonatal prophylactic antibiotics after preterm birth affect plasma proteome and immune development in pigs. Pediatr Res 94, 530–538 (2023). https://doi.org/10.1038/s41390-023-02492-7
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DOI: https://doi.org/10.1038/s41390-023-02492-7
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