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Am J Perinatol 2023; 40(08): 817-824
DOI: 10.1055/s-0043-1761916
SMFM Fellowship Series Article

The Effects of Pregnancy on the Pulmonary Immune Response in a Mouse Model of LPS-Induced Acute Lung Injury

Rebecca E. Rieck
1   Division of Maternal Fetal Medicine, Department of Obstetrics and Gynecology, School of Medicine, University of Virginia, Charlottesville, Virginia
,
Joseph J. Bivona III
2   Department of Medicine, Larner College of Medicine, University of Vermont, Burlington, Vermont
3   Cellular, Molecular, and Biomedical Sciences Doctoral Program, University of Vermont, Burlington, Vermont
,
Laura R. Hoyt
4   Department of Immunobiology, School of Medicine, Yale University, New Haven, Connecticut
,
Sebastian Ventrone
2   Department of Medicine, Larner College of Medicine, University of Vermont, Burlington, Vermont
,
Marta Kokoszynska
2   Department of Medicine, Larner College of Medicine, University of Vermont, Burlington, Vermont
,
Elizabeth A. Bonney
5   Department of Obstetrics, Gynecology, and Reproductive Sciences, Larner College of Medicine, Burlington, Vermont
,
Benjamin T. Suratt
2   Department of Medicine, Larner College of Medicine, University of Vermont, Burlington, Vermont
› Author Affiliations Funding Support for this work came from the Mead Fund, and the Department of Obstetrics, Gynecology and Reproductive Sciences at the Larner College of Medicine at the University of Vermont.
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Abstract

Objective This study evaluated the effect of pregnancy on the pulmonary innate immune response in a mouse model of acute lung injury (ALI) using nebulized lipopolysaccharide (LPS).

Study Design Pregnant (day 14) C57BL/6NCRL mice and nonpregnant controls received nebulized LPS for 15 minutes. Twenty-four hours later, mice were euthanized for tissue harvest. Analysis included blood and bronchoalveolar lavage fluid (BALF) differential cell counts, whole-lung inflammatory cytokine transcription levels by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR), and whole-lung vascular cell adhesion molecule 1 (VCAM-1), intercellular adhesion molecule 1 (ICAM-1), and BALF albumin by western blot. Mature bone marrow neutrophils from uninjured pregnant and nonpregnant mice were examined for chemotactic response using a Boyden chamber and for cytokine response to LPS by RT-qPCR.

Results In LPS-induced ALI, pregnant mice had higher BALF total cell (p < 0.001) and neutrophil counts (p < 0.001) as well as higher peripheral blood neutrophils (p < 0.01) than nonpregnant mice, but a similar increase (as compared with unexposed mice) in airspace albumin levels. Whole-lung expression of interleukin 6, tumor necrosis factor-α (TNF-α), and keratinocyte chemoattractant (CXCL1) was also similar. In vitro, marrow-derived neutrophils from pregnant and nonpregnant mice had similar chemotaxis to CXCL1 and N-formylmethionine-leucyl-phenylalanine, but neutrophils from pregnant mice expressed lower levels of TNF (p < 0.001) and CXCL1 (p < 0.01) after LPS stimulation. In uninjured mice, VCAM-1 was higher in lungs from pregnant versus nonpregnant mice (p < 0.05).

Conclusion In this model, pregnancy is associated with an augmented lung neutrophil response to ALI without increased capillary leak or whole-lung cytokine levels relative to the nonpregnant state. This may stem from increased peripheral blood neutrophil response and intrinsically increased expression of pulmonary vascular endothelial adhesion molecules. Differences in lung innate cell homeostasis may affect the response to inflammatory stimuli and explain severe lung disease in respiratory infection during pregnancy.

Key Points

  • Inhalation of LPS in midgestation versus virgin mice is associated with increased neutrophilia.

  • This occurs without a comparative increase in cytokine expression.

  • This may be explained by pregnancy-enhanced pre-exposure expression of VCAM-1 and ICAM-1.

Keywords

pulmonary immune - maternal lung - mouse model - lung injury

Supplementary Material



Publication History

Received: 19 November 2021

Accepted: 24 December 2022

Article published online:
16 February 2023

© 2023. Thieme. All rights reserved.

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  • References

  • 1 Rush B, Martinka P, Kilb B, McDermid RC, Boyd JH, Celi LA. Acute respiratory distress syndrome in pregnant women. Obstet Gynecol 2017; 129 (03) 530-535
    CrossrefPubMedGoogle Scholar
  • 2 Mabie WC, Barton JR, Sibai BM. Adult respiratory distress syndrome in pregnancy. Am J Obstet Gynecol 1992; 167 (4 Pt 1): 950-957
    CrossrefPubMedGoogle Scholar
  • 3 Muthu V, Agarwal R, Dhooria S. et al. Epidemiology, lung mechanics and outcomes of ARDS: a comparison between pregnant and non-pregnant subjects. J Crit Care 2019; 50: 207-212
    CrossrefPubMedGoogle Scholar
  • 4 Siston AM, Rasmussen SA, Honein MA. et al; Pandemic H1N1 Influenza in Pregnancy Working Group. Pandemic 2009 influenza A(H1N1) virus illness among pregnant women in the United States. JAMA 2010; 303 (15) 1517-1525
    CrossrefPubMedGoogle Scholar
  • 5 ANZIC Influenza Investigators and Australasian Maternity Outcomes Surveillance System. Critical illness due to 2009 A/H1N1 influenza in pregnant and postpartum women: population based cohort study. BMJ 2010; 340: c1279
    CrossrefPubMedGoogle Scholar
  • 6 Metz TD, Clifton RG, Hughes BL. et al; Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD) Maternal-Fetal Medicine Units (MFMU) Network. Disease severity and perinatal outcomes of pregnant patients with coronavirus disease 2019 (COVID-19). Obstet Gynecol 2021; 137 (04) 571-580
    CrossrefPubMedGoogle Scholar
  • 7 Zambrano LD, Ellington S, Strid P. et al; CDC COVID-19 Response Pregnancy and Infant Linked Outcomes Team. Update: characteristics of symptomatic women of reproductive age with laboratory-confirmed SARS-CoV-2 infection by pregnancy status - United States, January 22-October 3, 2020. MMWR Morb Mortal Wkly Rep 2020; 69 (44) 1641-1647
    CrossrefPubMedGoogle Scholar
  • 8 Marcelin G, Aldridge JR, Duan S. et al. Fatal outcome of pandemic H1N1 2009 influenza virus infection is associated with immunopathology and impaired lung repair, not enhanced viral burden, in pregnant mice. J Virol 2011; 85 (21) 11208-11219
    CrossrefPubMedGoogle Scholar
  • 9 Chan KH, Zhang AJX, To KKW. et al. Wild type and mutant 2009 pandemic influenza A (H1N1) viruses cause more severe disease and higher mortality in pregnant BALB/c mice. PLoS One 2010; 5 (10) e13757
    CrossrefPubMedGoogle Scholar
  • 10 Engels G, Hierweger AM, Hoffmann J. et al. Pregnancy-related immune adaptation promotes the emergence of highly virulent H1N1 influenza virus strains in allogenically pregnant mice. Cell Host Microbe 2017; 21 (03) 321-333
    CrossrefPubMedGoogle Scholar
  • 11 Matute-Bello G, Downey G, Moore BB. et al; Acute Lung Injury in Animals Study Group. An official American Thoracic Society workshop report: features and measurements of experimental acute lung injury in animals. Am J Respir Cell Mol Biol 2011; 44 (05) 725-738
    CrossrefPubMedGoogle Scholar
  • 12 Huffman LJ, Frazer DG, Prugh DJ. et al. Enhanced pulmonary inflammatory response to inhaled endotoxin in pregnant rats. J Toxicol Environ Health A 2004; 67 (02) 125-144
    CrossrefPubMedGoogle Scholar
  • 13 Kordonowy LL, Burg E, Lenox CC. et al. Obesity is associated with neutrophil dysfunction and attenuation of murine acute lung injury. Am J Respir Cell Mol Biol 2012; 47 (01) 120-127
    CrossrefPubMedGoogle Scholar
  • 14 Ubags ND, Vernooy JH, Burg E. et al. The role of leptin in the development of pulmonary neutrophilia in infection and acute lung injury. Crit Care Med 2014; 42 (02) e143-e151
    CrossrefPubMedGoogle Scholar
  • 15 Ubags NDJ, Stapleton RD, Vernooy JHJ. et al. Hyperleptinemia is associated with impaired pulmonary host defense. JCI Insight 2016; 1 (08) e82101
    PubMedGoogle Scholar
  • 16 Rudmann DG, Moore MW, Tepper JS. et al. Modulation of allergic inflammation in mice deficient in TNF receptors. Am J Physiol Lung Cell Mol Physiol 2000; 279 (06) L1047-L1057
    CrossrefPubMedGoogle Scholar
  • 17 Petty JM, Sueblinvong V, Lenox CC. et al. Pulmonary stromal-derived factor-1 expression and effect on neutrophil recruitment during acute lung injury. J Immunol 2007; 178 (12) 8148-8157
    CrossrefPubMedGoogle Scholar
  • 18 Suratt BT, Young SK, Lieber J, Nick JA, Henson PM, Worthen GS. Neutrophil maturation and activation determine anatomic site of clearance from circulation. Am J Physiol Lung Cell Mol Physiol 2001; 281 (04) L913-L921
    CrossrefPubMedGoogle Scholar
  • 19 Suratt BT, Petty JM, Young SK. et al. Role of the CXCR4/SDF-1 chemokine axis in circulating neutrophil homeostasis. Blood 2004; 104 (02) 565-571
    CrossrefPubMedGoogle Scholar
  • 20 Madenspacher JH, Draper DW, Smoak KA. et al. Dyslipidemia induces opposing effects on intrapulmonary and extrapulmonary host defense through divergent TLR response phenotypes. J Immunol 2010; 185 (03) 1660-1669
    CrossrefPubMedGoogle Scholar
  • 21 Nourshargh S, Alon R. Leukocyte migration into inflamed tissues. Immunity 2014; 41 (05) 694-707
    CrossrefPubMedGoogle Scholar
  • 22 Austgulen R, Lien E, Vince G, Redman CWG. Increased maternal plasma levels of soluble adhesion molecules (ICAM-1, VCAM-1, E-selectin) in preeclampsia. Eur J Obstet Gynecol Reprod Biol 1997; 71 (01) 53-58
    CrossrefPubMedGoogle Scholar
  • 23 Krauss T, Kuhn W, Lakoma C, Augustin HG. Circulating endothelial cell adhesion molecules as diagnostic markers for the early identification of pregnant women at risk for development of preeclampsia. Am J Obstet Gynecol 1997; 177 (02) 443-449
    CrossrefPubMedGoogle Scholar
  • 24 Lewis DF, Canzoneri BJ, Gu Y, Zhao S, Wang Y. Maternal levels of prostacyclin, thromboxane, ICAM, and VCAM in normal and preeclamptic pregnancies. Am J Reprod Immunol 2010; 64 (06) 376-383
    CrossrefPubMedGoogle Scholar
  • 25 Crouch SP, Crocker IP, Fletcher J. The effect of pregnancy on polymorphonuclear leukocyte function. J Immunol 1995; 155 (11) 5436-5443
    CrossrefPubMedGoogle Scholar
  • 26 Crocker IP, Baker PN, Fletcher J. Neutrophil function in pregnancy and rheumatoid arthritis. Ann Rheum Dis 2000; 59 (07) 555-564
    CrossrefPubMedGoogle Scholar
  • 27 Krause PJ, Ingardia CJ, Pontius LT, Malech HL, LoBello TM, Maderazo EG. Host defense during pregnancy: neutrophil chemotaxis and adherence. Am J Obstet Gynecol 1987; 157 (02) 274-280
    CrossrefPubMedGoogle Scholar
  • 28 Robson SC, Hunter S, Boys RJ, Dunlop W. Serial changes in pulmonary haemodynamics during human pregnancy: a non-invasive study using Doppler echocardiography. Clin Sci (Lond) 1991; 80 (02) 113-117
    CrossrefPubMedGoogle Scholar
  • 29 Fay ME, Myers DR, Kumar A. et al. Cellular softening mediates leukocyte demargination and trafficking, thereby increasing clinical blood counts. Proc Natl Acad Sci U S A 2016; 113 (08) 1987-1992
    CrossrefPubMedGoogle Scholar
  • 30 Doerschuk CM, Allard MF, Lee S, Brumwell ML, Hogg JC. Effect of epinephrine on neutrophil kinetics in rabbit lungs. J Appl Physiol 1988; 65 (01) 401-407
    CrossrefPubMedGoogle Scholar
  • 31 Jeyaseelan S, Chu HW, Young SK, Worthen GS. Transcriptional profiling of lipopolysaccharide-induced acute lung injury. Infect Immun 2004; 72 (12) 7247-7256
    CrossrefPubMedGoogle Scholar
  • 32 Salminen A, Paananen R, Vuolteenaho R. et al. Maternal endotoxin-induced preterm birth in mice: fetal responses in toll-like receptors, collectins, and cytokines. Pediatr Res 2008; 63 (03) 280-286
    CrossrefPubMedGoogle Scholar
  • 33 Barnes BJ, Adrover JM, Baxter-Stoltzfus A. et al. Targeting potential drivers of COVID-19: Neutrophil extracellular traps. J Exp Med 2020; 217 (06) 1-7
    CrossrefPubMedGoogle Scholar
  • 34 Vardhana SA, Wolchok JD. The many faces of the anti-COVID immune response. J Exp Med 2020; 217 (06) 1-10
    CrossrefPubMedGoogle Scholar