European Journal of Nutrition

, Volume 56, Issue 2, pp 693–704 | Cite as

Relation between neonatal malnutrition and gene expression: inflammasome function in infections caused by Candida Albicans

  • Thacianna Barreto Da Costa
  • Natália Gomes De Morais
  • Joana Maria Bezerra De Lira
  • Thays Miranda De Almeida
  • Suênia Da Cunha Gonçalves-De-Albuquerque
  • Valéria Rêgo Alves Pereira
  • Milena De Paiva Cavalcanti
  • Célia Maria Machado Barbosa De Castro
Original Contribution



To investigate the effects of neonatal malnutrition followed by nutritional replacement on the signaling mechanisms developed by the inflammasome complex by analyzing the expression of the targeted TLR2, TLR4, NLRP3, caspase-1 and release of IL-1β and IL-18 by alveolar macrophages infected in vitro with Candida albicans.


Male Wistar rats (n = 24), 90–120 days, were suckled by mothers whose diet during lactation contained 17 % protein in the nourish group and 8 % protein in the malnourished group. After weaning, both groups were fed a normal protein diet. Macrophages were obtained after tracheostomy, through the collection of bronchoalveolar lavage fluid. The quantification of the expression levels of targets (TLR2, TLR4, NLRP3 and caspase-1) was performed by real-time RT-PCR. Production of cytokines was performed by ELISA.


The malnourished animals during lactation showed reduced body weight from the fifth day of life, remaining until adulthood. Further, the model applied malnutrition induced a lower expression of TLR4 and caspase-1. The quantification of the TLR2 and NLRP3, as well as the release of IL-1β and IL-18, was not different between groups of animals nourished and malnourished. The system challenged with Candida albicans showed high expression levels of all targets in the study.


The tests demonstrate nutritional restriction during critical periods of development, although nutritional supplementation may compromise defense patterns in adulthood in a timely manner, preserving distinct signaling mechanism, so that the individual does not become widely vulnerable to infections by opportunistic pathogens.


Neonatal malnutrition Programming Macrophage Candida albicans Toll-like receptors NOD-like receptors 



The authors wish to thank the Keizo Asami Immunopathology Laboratory of the Federal University of Pernambuco, Recife, Brazil, and Department of Immunology, Aggeu Magalhães Research Center, Oswaldo Cruz Foundation, Campus UFPE, Recife, Brazil, for the structure and equipment provided for the development of this research.


The study was supported by National Counsel of Technological and Scientific Development (CNPq), Brazil, and partially funded by Dean of Research and Graduate Studies, Federal University of Pernambuco (PROPESQ/UFPE), Brazil.

Compliance with ethical standards

Conflict of interest

On behalf of all authors, the corresponding author states that there is no conflict of interest.

Ethical standards

This study was approved by Ethics Committee on Animal Experimentation of the Center for Biological Sciences, Federal University of Pernambuco (CEEA-UFPE) (Pernambuco, Brazil, Protocol 23076.053096/2011-91 ). The manuscript does not contain clinical studies or patient data.


  1. 1.
    Laus MF, Vales LDMF, Costa TMB, Almeida SS (2011) Early postnatal protein-calorie malnutrition and cognition: a review of human and animal studies. Int J Environ Res Public Health 8:590–612CrossRefGoogle Scholar
  2. 2.
    Lucas A (2005) Long-term programming effects of early nutrition—implications for the preterm infant. J Perinatol 25(Suppl 2):S2–S6CrossRefGoogle Scholar
  3. 3.
    Hausman DB, McCloskey HM, Martin RJ (1991) Maternal fat type influences the growth and fatty acid composition of newborn and weanling rats. J Nutr 121:1917–1923Google Scholar
  4. 4.
    Kozak R, Burle A, Burle C, Beck B (2000) Dietary composition during fetal and neonatal life affects neuropeptide Y functioning in adult offspring. Dev Brain Res 125:75–82CrossRefGoogle Scholar
  5. 5.
    Cunningham-Rundles S, McNeeley DF, Moon A (2005) Mechanisms of nutrient modulation of the immune response. J Allergy Clin Immunol 115:1119–1128CrossRefGoogle Scholar
  6. 6.
    Marcos A, Nova E, Montero A (2003) Changes in the immune system are conditioned by nutrition. Eur J Clin Nutr 57(Suppl 1):S66–S69CrossRefGoogle Scholar
  7. 7.
    Palmer AC (2011) Nutritionally mediated programming of the developing immune system. Adv Nutr 2:377–395. doi: 10.3945/an.111.000570 CrossRefGoogle Scholar
  8. 8.
    Costa TB, Morais NG, Almeida TM, Severo MS, De Castro CMMB (2012) Early malnutrition and production of IFN-γ, IL-12 and IL-10 by macrophages/lymphocytes: in vitro study of cell infection by methicillin-sensitive and methicillin-resistant Staphylococcus aureus. Rev Nutr 25:607–619CrossRefGoogle Scholar
  9. 9.
    Melo JF, Costa TB, Lima TDC, Chaves MEC, Vayssade LM, Nagel MD, Castro CMMB (2012) Long-term effects of a neonatal low-protein diet in rats on the number of macrophages in culture and the expression/production of fusion proteins. Eur J Nutr 52:1475–1482CrossRefGoogle Scholar
  10. 10.
    Meyer J, Hinder F, Stothert J Jr et al (1994) Increased organ blood flow in chronic endotoxemia is reversed by nitric oxide synthase inhibition. J Appl Physiol 76:2785–2793Google Scholar
  11. 11.
    Amersfoort ESV, Berkel TJCV, Kuiper J (2003) Receptors, mediators, and mechanisms involved in bacterial sepsis and septic shock. Clin Microbiol Rev 16:379–414CrossRefGoogle Scholar
  12. 12.
    Scrimshaw NS, SanGiovanni JP (1997) Synergism of nutrition, infection, and immunity: an overview. Am J Clin Nutr 66:464S–477SGoogle Scholar
  13. 13.
    Gordon JE (1976) Synergism of malnutrition and infectious disease. In: Beaton GH, Bengoa JM (eds) Nutrition in preventive medicine. WHO, Geneva, pp 193–209. (WHO Monograph Series, 62)Google Scholar
  14. 14.
    Trick WE (2002) Secular trend of hospital-acquired candidemia among intensive care unit patients in the United States during 1989–99. Clin Infect Dis 35:627–630CrossRefGoogle Scholar
  15. 15.
    LeibundGut-Landmann S, Wüthrich M, Hohl MT (2012) Immunity to Fungi. Curr Opin Immunol 24:449–458CrossRefGoogle Scholar
  16. 16.
    Ifrim DC, Joosten LA, Kullberg BJ, Jacobs L, Jansen T, Williams DL, Gow NA, van der Meer JW, Netea MG, Quintin J (2013) Candida albicans primes TLR cytokine responses through a Dectin-1/Raf-1-mediated pathway. J Immunol 190:4129–4135CrossRefGoogle Scholar
  17. 17.
    Gil ML, Gozalbo D (2006) TLR2, but not TLR4, triggers cytokine production by murine cells in response to Candida albicans yeasts and hyphae. Microbes Infect 8:2299–2304CrossRefGoogle Scholar
  18. 18.
    Netea MG, Van Der Graaf CA, Vonk AG, Verschueren I, Van Der Meer JWM, Kulberg BJ (2002) The role of Toll-like receptor (TLR) 2 and TLR 4 in the host defense against disseminated candidiasis. J Infect Dis 185:1483–1489CrossRefGoogle Scholar
  19. 19.
    Hise AG, Tomalka J, Ganesan S, Patel K, Hall BA, Brown GD, Fitzgerald KA (2009) An essential role for the NLRP3 inflammasome in host defense against the human fungal pathogen, Candida albicans. Cell Host Microbe 5:487–497CrossRefGoogle Scholar
  20. 20.
    Gross O, Poeck H, Bscheider M et al (2009) Syk kinase signalling couples to the Nlrp3 inflammasome for anti-fungal host defence. Nature 459:433–436CrossRefGoogle Scholar
  21. 21.
    Godfrey KM, Barker DJ (2001) Fetal programming and adult health. Public Health Nutr 4:611–624CrossRefGoogle Scholar
  22. 22.
    Seymonds ME, Budge H, Stephenson T, Gardner DS (2005) Experimental evidence for long-term programming effects of early diet. Adv Exp Med Biol 569:24–32CrossRefGoogle Scholar
  23. 23.
    Pico C, Palou A (2013) Perinatal programming of obesity: an introduction to the topic. Front Physiol 4:255CrossRefGoogle Scholar
  24. 24.
    De Castro CMMB, Castro RM, Medeiros AF, Santos AQ, Ferreira-e-Silva WT, Lima Filho JL (2000) Effect of stress on the production of O2-in alveolar macrophages. J Neuroimmunol 108:68–72CrossRefGoogle Scholar
  25. 25.
    Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739CrossRefGoogle Scholar
  26. 26.
    Sharrocks AD (1994) The design of primers for PCR, vol 2. CRC Press Inc, Boca Raton, pp 5–10Google Scholar
  27. 27.
    Chiba T, Itoh T, Tabuchi M, Nakazawa T, Satou T (2012) Interleukin-1β accelerates the onset of stroke in stroke-prone spontaneously hypertensive rats. Mediators Inflamm 2012:701976CrossRefGoogle Scholar
  28. 28.
    Too HP (2003) Real time PCR quantification of GFR-2 alternatively spliced isoforms in murine brain and peripheral tissues. Brain Res Mol Brain Res 114:146–154CrossRefGoogle Scholar
  29. 29.
    Kubista M, Andrade JM, Bengtsson M, Forootan A, Jonák J, Lind K, Sindelka R, Sjöback R, Sjögreen B, Strömbom L et al (2006) The real time polymerase chain reaction. Mol Aspects Med 27:95–125CrossRefGoogle Scholar
  30. 30.
    Gonçalves SC, Régis-da-Silva CG, Brito MEF, Brandão-Filho SP, Paiva-Cavalcanti M (2012) Application of the mammalian glyceraldehyde-3-phosphate dehydrogenase gene for sample quality control in multiplex PCR for diagnosis of leishmaniasis. J Venom Anim Toxins Incl Trop Dis 18:188–197CrossRefGoogle Scholar
  31. 31.
    World Health Organization (WHO) lactation. (1994) In: World Health Organization (WHO) Healthy Children physiological bases. São Paulo (SP): IBFAN Brazil and Health Institute, OMS, OPAS e UNICEF Brasil, pp 17–35Google Scholar
  32. 32.
    Araujo FRG, De Castro CMMB, Rocha JA, Sampaio B, Diniz MFA, Eve LB, Montarroyos UR (2012) Perialveolar bacterial microbiota and bacteraemia after dental alveolitis in adult rats that had been subjected to neonatal malnutrition. Br J Nutr 107:996–1005CrossRefGoogle Scholar
  33. 33.
    Passos MC, da Fonte Ramos C, Dutra SC, Mouco T, de Moura EG (2002) Long-term effects of malnutrition during lactation on the thyroid function of offspring. Horm Metab Res 34:40–43CrossRefGoogle Scholar
  34. 34.
    Ngom PT, Collinson AC, Pido-Lopez J, Henson SM, Prentice AM, Aspinall R (2004) Improved thymic function in exclusively breastfed infants is associated with higher interleukin 7 concentrations in their mothers’ breast milk. Am J Clin Nutr 80:722–728Google Scholar
  35. 35.
    Bittencourt FB (2010) Os efeitos da restrição alimentar materna durante o período de lactação em diferentes sistemas orgânicos da prole em ratos. Acta Scientiae Medica 3:26–33Google Scholar
  36. 36.
    Sayer AA, Cooper C (2005) Fetal programming of body com- position and musculoskeletal development. Early Hum Dev 81:735–744CrossRefGoogle Scholar
  37. 37.
    Gallou-Kabani C, Junien C (2005) Nutritional epigenomics of metabolic syndrome. New perspective against the epidemic. Diabetes 54:1899–1906CrossRefGoogle Scholar
  38. 38.
    Redmond HP, Shou J, Kelly CJ, Schreiber S, Miller E, Leon P, Daly JM (1991) Immunosuppressive mechanisms in protein-calorie malnutrition. Surgery 110:311–317Google Scholar
  39. 39.
    Redmond HP, Shou J, Kelly CJ, Leon P, Daly JM (1991) Protein-calorie malnutrition impairs host defense against Candida albicans. J Surg Res 50:552–559CrossRefGoogle Scholar
  40. 40.
    Brunetto MA, Gomes MOS, Jeremias JT, Oliveira LD, Carciofi AC (2007) Imunonutrição: O papel da dieta no restabelecimento das defesas naturais. Acta Scien Veterinariae 35:5230–5232Google Scholar
  41. 41.
    Fock RA, Vinolo MA, de Moura Sá Rocha V, de Sá Rocha LC, Borelli P (2007) Protein-energy malnutrition decreases the expression of TLR-4/MD-2 and CD14 receptors in peritoneal macrophage and reduces the synthesis of TNF-alpha in response to lipopolysaccharide (LPS) in mice. Cytokine 40:105–114CrossRefGoogle Scholar
  42. 42.
    Morais NG, Costa TB, Severo MS, De Castro CMMB (2014) Long-term effects of a neonatal malnutrition on the microbicide response, production of cytokines and survival of macrophages infected by Staphylococcus aureus sensitive/resistant to methicillin. Nutr J 27:557–568Google Scholar
  43. 43.
    Wellington M, Koselny K, Sutterwala FS, Krysan DJ (2014) Candida albicans triggers NLRP3-mediated pyroptosis in macrophages. Eukaryot Cell 13:329–340CrossRefGoogle Scholar
  44. 44.
    Azevedo ZMA, Luz RA, Victal SH, Kurdian B, Fonseca VM, Fitting C, Cmara FP, Haeffner-Cavaillon N, Cavaillon JM, Gaspar Elsas MIC, Kavier Elsas P (2005) Increased production of tumor necrosis factor-α in whole blood cultures from children with primary malnutrition. Braz J Med Biol Res 38:171–183CrossRefGoogle Scholar
  45. 45.
    Netea MG, Simon A, van de Veerdonk F, Kullberg BJ, Van der Meer JW, Joosten LA (2009) IL-1beta processing in host defense: beyond the inflammasomes. PLoS Pathog 6:e1000661CrossRefGoogle Scholar
  46. 46.
    Rossi T, Lozovoy MAB, Silva RV et al (2011) Interactions between Candida albicans and host. Semina Ciências Biológicas e da Saúde 32:15–28CrossRefGoogle Scholar
  47. 47.
    Cheng SC, Joosten LA, Kullberg BJ, Netea MG (2012) Interplay between Candida albicans and the mammalian innate host defense. Infect Immun 80:1304–1313CrossRefGoogle Scholar
  48. 48.
    Desroche N, Beltramo C, Guzzo J (2005) Determination of an internal control to apply reverse transcription quantitative PCR to study stress response in the lactic acid bacterium Oenococcus oeni. J Microbiol Methods 60:325–333CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Thacianna Barreto Da Costa
    • 1
    • 2
  • Natália Gomes De Morais
    • 1
    • 2
  • Joana Maria Bezerra De Lira
    • 2
  • Thays Miranda De Almeida
    • 3
  • Suênia Da Cunha Gonçalves-De-Albuquerque
    • 3
  • Valéria Rêgo Alves Pereira
    • 3
  • Milena De Paiva Cavalcanti
    • 3
  • Célia Maria Machado Barbosa De Castro
    • 1
    • 2
  1. 1.Department of Tropical MedicineFederal University of PernambucoRecifeBrazil
  2. 2.Keizo Asami Laboratory of ImmunopathologyFederal University of PernambucoRecifeBrazil
  3. 3.Department of Immunology, Aggeu Magalhães Research CenterOswaldo Cruz FoundationRecifeBrazil

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