Characterization of regulatory T cells in urban newborns

Ngoc P Ly, Begona Ruiz-Perez, Rachel M McLoughlin, Cynthia M Visness, Paul K Wallace, William W Cruikshank, Arthur O Tzianabos, George T O'Connor, Diane R Gold, James E Gern, Ngoc P Ly, Begona Ruiz-Perez, Rachel M McLoughlin, Cynthia M Visness, Paul K Wallace, William W Cruikshank, Arthur O Tzianabos, George T O'Connor, Diane R Gold, James E Gern

Abstract

Background: In the United States, asthma prevalence is particularly high among urban children. Although the underlying immune mechanism contributing to asthma has not been identified, having impaired T regulatory (Treg) cells at birth may be a determining factor in urban children. The objective of this study was to compare Treg phenotype and function in cord blood (CB) of newborns to those in peripheral blood (PB) of a subset of participating mothers.

Methods: Treg numbers, expression, and suppressive function were quantified in subjects recruited prenatally from neighborhoods where >/= 20% of families have incomes below the poverty line. Proportion of Treg cells and expression of naïve (CD45RA) or activated (CD45RO, CD69, and HLA-DR) markers in CD4+T cells was measured by flow cytometry. Treg suppressive capacity was determined by quantifying PHA-stimulated lymphocyte proliferation in mononuclear cell samples with and without CD25 depletion.

Results: In an urban cohort of 119 newborns and 82 mothers, we found that newborns had similar number of cells expressing FOXP3 as compared to the mothers but had reduced numbers of CD4+CD25+bright cells that predominantly expressed the naïve (CD45RA) rather than the activated/memory (CD45RO) phenotype found in the mothers. Additionally, the newborns had reduced mononuclear cell TGF-beta production, and reduced Treg suppression of PHA-stimulated lymphocyte proliferation compared to the mothers.

Conclusion: U.S. urban newborns have Treg cells that express FOXP3, albeit with an immature phenotype and function as compared to the mothers. Longitudinal follow-up is needed to delineate Treg cell maturation and subsequent risk for atopic diseases in this urban birth cohort.

Figures

Figure 1
Figure 1
TGF-β secretion in mononuclear cells and CD25 and Foxp3 expression in CD4+ T-cells of cord blood (CB) and peripheral blood (PB). (A) Contour plots of CD4 and CD25 expression in unstimulated CB and PB T-cells. Representative examples of one out of 114 CB and 79 PB samples analyzed are shown, illustrating the separation of the CD25- and CD25+ populations in CB CD4+ cells as compared to a broader range CD25 expression in PB CD4+ cells. (B) Production of TGF-β cytokines by CB (n = 49) and PB (n = 59) mononuclear cells (MNCs) measured by ELISA in supernatants 4 days after incubation in media (unstimulated) and phytohemagglutinin (PHA). The median is represented by the horizontal bar within the box. The upper and lower boundaries of the box represent the 25th to 75th percentiles of the data, respectively. Observations < 1.5 times the height of the box beyond either quartile are displayed within the whiskers. (C) Intracellular expression of Foxp3 in unstimulated samples of CB and PB MNCs analyzed by flow cytometry. The CD4+ cells were gated and analyzed for expression of CD25 and FOXP3. The percentage of CD4+ cells expressing CD25 and FOXP3 is shown in the upper right-hand quadrants. FOXP3 are not distinctly expressed within the CD4+CD25+bright cell population in CB as compared to PB. Compared to CB, maternal PB had a significant population of CD25+FOXP3- cells (upper left-hand quadrants). Results are representative examples of one out of 63 CB and 78 PB samples analyzed.
Figure 2
Figure 2
Comparison of activation markers between cord and peripheral blood CD4+ CD25+bright cells. CD45RO, CD45RA, CD69, and HLA-DR expression on CD4+ CD25+bright cells sorted by flow cytometry and expressed in percent. A majority of CB CD4+CD25+bright cells exhibited a naïve phenotype. In contrast, PB CD4+CD25+bright exhibited an activated/memory phenotype.

References

    1. Suto A, Nakajima H, Kagami SI, Suzuki K, Saito Y, Iwamoto I. Role of CD4(+) CD25(+) regulatory T cells in T helper 2 cell-mediated allergic inflammation in the airways. Am J Respir Crit Care Med. 2001;164:680–687.
    1. Zuany-Amorim C, Sawicka E, Manlius C, Le Moine A, Brunet LR, Kemeny DM, Bowen G, Rook G, Walker C. Suppression of airway eosinophilia by killed Mycobacterium vaccae-induced allergen-specific regulatory T-cells. Nat Med. 2002;8:625–629. doi: 10.1038/nm0602-625.
    1. Strickland DH, Stumbles PA, Zosky GR, Subrata LS, Thomas JA, Turner DJ, Sly PD, Holt PG. Reversal of airway hyperresponsiveness by induction of airway mucosal CD4+CD25+ regulatory T cells. J Exp Med. 2006;203:2649–2660. doi: 10.1084/jem.20060155.
    1. Kearley J, Barker JE, Robinson DS, Lloyd CM. Resolution of airway inflammation and hyperreactivity after in vivo transfer of CD4+CD25+ regulatory T cells is interleukin 10 dependent. J Exp Med. 2005;202:1539–1547. doi: 10.1084/jem.20051166.
    1. Maggi L, Santarlasci V, Liotta F, Frosali F, Angeli R, Cosmi L, Maggi E, Romagnani S, Annunziato F. Demonstration of circulating allergen-specific CD4+CD25highFoxp3+ T-regulatory cells in both nonatopic and atopic individuals. J Allergy Clin Immunol. 2007;120:429–436. doi: 10.1016/j.jaci.2007.05.002.
    1. Bellinghausen I, Klostermann B, Knop J, Saloga J. Human CD4+CD25+ T cells derived from the majority of atopic donors are able to suppress TH1 and TH2 cytokine production. J Allergy Clin Immunol. 2003;111:862–868. doi: 10.1067/mai.2003.1412.
    1. Ling EM, Smith T, Nguyen XD, Pridgeon C, Dallman M, Arbery J, Carr VA, Robinson DS. Relation of CD4+CD25+ regulatory T-cell suppression of allergen-driven T-cell activation to atopic status and expression of allergic disease. Lancet. 2004;363:608–615. doi: 10.1016/S0140-6736(04)15592-X.
    1. Grindebacke H, Wing K, Andersson AC, Suri-Payer E, Rak S, Rudin A. Defective suppression of Th2 cytokines by CD4CD25 regulatory T cells in birch allergics during birch pollen season. Clin Exp Allergy. 2004;34:1364–1372. doi: 10.1111/j.1365-2222.2004.02067.x.
    1. Hartl D, Koller B, Mehlhorn AT, Reinhardt D, Nicolai T, Schendel DJ, Griese M, Krauss-Etschmann S. Quantitative and functional impairment of pulmonary CD4+CD25hi regulatory T cells in pediatric asthma. J Allergy Clin Immunol. 2007;119:1258–1266. doi: 10.1016/j.jaci.2007.02.023.
    1. Sherman CB, Tosteson TD, Tager IB, Speizer FE, Weiss ST. Early childhood predictors of asthma. American Journal of Epidemiology. 1990;132:83–95.
    1. Yunginger JW, Reed CE, O'Connell EJ, Melton LJ, 3rd, O'Fallon WM, Silverstein MD. A community-based study of the epidemiology of asthma. Incidence rates, 1964–1983. Am Rev Respir Dis. 1992;146:888–894.
    1. Haddeland U, Karstensen AB, Farkas L, Bo KO, Pirhonen J, Karlsson M, Kvavik W, Brandtzaeg P, Nakstad B. Putative regulatory T cells are impaired in cord blood from neonates with hereditary allergy risk. Pediatr Allergy Immunol. 2005;16:104–112. doi: 10.1111/j.1399-3038.2005.00250.x.
    1. Schaub B, Liu J, Hoppler S, Haug S, Sattler C, Lluis A, Illi S, von Mutius E. Impairment of T-regulatory cells in cord blood of atopic mothers. J Allergy Clin Immunol. 2008;121:1491–1499. doi: 10.1016/j.jaci.2008.04.010.
    1. Litonjua AA, Carey VJ, Burge HA, Weiss ST, Gold DR. Parental history and the risk for childhood asthma. Does mother confer more risk than father? Am J Respir Crit Care Med. 1998;158:176–181.
    1. Cullinan P, MacNeill SJ, Harris JM, Moffat S, White C, Mills P, Newman Taylor AJ. Early allergen exposure, skin prick responses, and atopic wheeze at age 5 in English children: a cohort study. Thorax. 2004;59:855–861. doi: 10.1136/thx.2003.019877.
    1. Martinez FD, Wright AL, Taussig LM, Holberg CJ, Halonen M, Morgan WJ. Asthma and wheezing in the first six years of life. N Engl J Med. 1995;332:133–138. doi: 10.1056/NEJM199501193320301.
    1. Polk S, Sunyer J, Munoz-Ortiz L, Barnes M, Torrent M, Figueroa C, Harris J, Vall O, Anto JM, Cullinan P. A prospective study of Fel d1 and Der p1 exposure in infancy and childhood wheezing. Am J Respir Crit Care Med. 2004;170:273–278. doi: 10.1164/rccm.200310-1348OC.
    1. Kuiper S, Muris JW, Dompeling E, Kester AD, Wesseling G, Knottnerus JA, van Schayck CP. Interactive effect of family history and environmental factors on respiratory tract-related morbidity in infancy. J Allergy Clin Immunol. 2007;120:388–395. doi: 10.1016/j.jaci.2007.03.038.
    1. Weiss KB, Gergen PJ, Crain EF. Inner-city asthma. The epidemiology of an emerging US public health concern. Chest. 1992;101:362S–367S.
    1. Gottlieb DJ, Beiser AS, O'Connor GT. Poverty, race, and medication use are correlates of asthma hospitalization rates. A small area analysis in Boston. Chest. 1995;108:28–35. doi: 10.1378/chest.108.1.28.
    1. Busse WW, Mitchell H. Addressing issues of asthma in inner-city children. J Allergy Clin Immunol. 2007;119:43–49. doi: 10.1016/j.jaci.2006.10.021.
    1. Shreffler WG, Visness CM, Burger M, Cruikshank WW, Lederman HM, de la Morena M, Grindle K, Calatroni A, Sampson HA, Gern JE. Standardization and performance evaluation of mononuclear cell cytokine secretion assays in a multicenter study. BMC Immunol. 2006;7:29. doi: 10.1186/1471-2172-7-29.
    1. Levings MK, Sangregorio R, Roncarolo MG. Human cd25(+)cd4(+) t regulatory cells suppress naive and memory T cell proliferation and can be expanded in vitro without loss of function. J Exp Med. 2001;193:1295–1302. doi: 10.1084/jem.193.11.1295.
    1. Jiang H, Chess L. An integrated view of suppressor T cell subsets in immunoregulation. J Clin Invest. 2004;114:1198–1208.
    1. Taams LS, Vukmanovic-Stejic M, Smith J, Dunne PJ, Fletcher JM, Plunkett FJ, Ebeling SB, Lombardi G, Rustin MH, Bijlsma JW, et al. Antigen-specific T cell suppression by human CD4+CD25+ regulatory T cells. Eur J Immunol. 2002;32:1621–1630. doi: 10.1002/1521-4141(200206)32:6<1621::AID-IMMU1621>;2-Q.
    1. Baecher-Allan C, Brown JA, Freeman GJ, Hafler DA. CD4+CD25high regulatory cells in human peripheral blood. J Immunol. 2001;167:1245–1253.
    1. R: A language and environment for statistical computing
    1. Chen W, Jin W, Hardegen N, Lei KJ, Li L, Marinos N, McGrady G, Wahl SM. Conversion of peripheral CD4+CD25- naive T cells to CD4+CD25+ regulatory T cells by TGF-beta induction of transcription factor Foxp3. J Exp Med. 2003;198:1875–1886. doi: 10.1084/jem.20030152.
    1. Khattri R, Cox T, Yasayko SA, Ramsdell F. An essential role for Scurfin in CD4+CD25+ T regulatory cells. Nat Immunol. 2003;4:337–342. doi: 10.1038/ni909.
    1. Sakaguchi S. Naturally arising CD4+ regulatory t cells for immunologic self-tolerance and negative control of immune responses. Annu Rev Immunol. 2004;22:531–562. doi: 10.1146/annurev.immunol.21.120601.141122.
    1. Fontenot JD, Rasmussen JP, Williams LM, Dooley JL, Farr AG, Rudensky AY. Regulatory T cell lineage specification by the forkhead transcription factor foxp3. Immunity. 2005;22:329–341. doi: 10.1016/j.immuni.2005.01.016.
    1. Takahata Y, Nomura A, Takada H, Ohga S, Furuno K, Hikino S, Nakayama H, Sakaguchi S, Hara T. CD25+CD4+ T cells in human cord blood: an immunoregulatory subset with naive phenotype and specific expression of forkhead box p3 (Foxp3) gene. Exp Hematol. 2004;32:622–629. doi: 10.1016/j.exphem.2004.03.012.
    1. Wing K, Ekmark A, Karlsson H, Rudin A, Suri-Payer E. Characterization of human CD25+ CD4+ T cells in thymus, cord and adult blood. Immunology. 2002;106:190–199. doi: 10.1046/j.1365-2567.2002.01412.x.
    1. Wing K, Larsson P, Sandstrom K, Lundin SB, Suri-Payer E, Rudin A. CD4+ CD25+ FOXP3+ regulatory T cells from human thymus and cord blood suppress antigen-specific T cell responses. Immunology. 2005;115:516–525. doi: 10.1111/j.1365-2567.2005.02186.x.
    1. Seddiki N, Santner-Nanan B, Tangye SG, Alexander SI, Solomon M, Lee S, Nanan R, Fazekas de Saint Groth B. Persistence of naive CD45RA+ regulatory T cells in adult life. Blood. 2006;107:2830–2838. doi: 10.1182/blood-2005-06-2403.
    1. Schaub B, Liu J, Schleich I, Hoppler S, Sattler C, von Mutius E. Impairment of T helper and T regulatory cell responses at birth. Allergy. 2008;63:1438–1447. doi: 10.1111/j.1398-9995.2008.01685.x.
    1. Allan SE, Crome SQ, Crellin NK, Passerini L, Steiner TS, Bacchetta R, Roncarolo MG, Levings MK. Activation-induced FOXP3 in human T effector cells does not suppress proliferation or cytokine production. Int Immunol. 2007;19:345–354. doi: 10.1093/intimm/dxm014.
    1. Gavin MA, Torgerson TR, Houston E, DeRoos P, Ho WY, Stray-Pedersen A, Ocheltree EL, Greenberg PD, Ochs HD, Rudensky AY. Single-cell analysis of normal and FOXP3-mutant human T cells: FOXP3 expression without regulatory T cell development. Proc Natl Acad Sci USA. 2006;103:6659–6664. doi: 10.1073/pnas.0509484103.

Source: PubMed

Sponzoři a spolupracovníci

Zdravotní podmínky

Drogové intervence

3
Předplatit