Abstract
Cell-mediated immunity (CMI) in neonates is relatively deficient when compared with adults. Defects in cytokine production and/or regulation may contribute to heightened susceptibility to infection by intracellular pathogens. The heterodimeric cytokine IL-12 is a key regulator of CMI and inducer of interferon-γ (IFN-γ) production. We report here that umbilical cord blood-derived mononuclear cells (MNC) are capable of producing IL-12 (p40 subunit, measured by RIA, and IL-12 p70 heterodimer, by ELISA) at levels comparable to or greater than adult peripheral blood MNC, after stimulation with heat-killed Staphylococcus aureus in 18-h cultures. As in adult MNC, S. aureus induced IL-12 p40 mRNA accumulation in cord blood MNC. IFN-γ was also produced in the S. aureus-stimulated cultures, in an IL-12-dependent manner, but cord blood MNC produced 5-fold lower levels of IFN-γ compared with adult MNC(p < 0.05). Preincubation with IL-10 inhibited IL-12 p40 production by cord blood and adult peripheral blood MNC in a dose-dependent fashion, whereas neutralization of endogenous IL-10 enhanced IL-12 and IFN-γ levels. The results demonstrate that the relative CMI deficiency in neonates is not due to an intrinsic defect in the capacity of neonatal MNC to produce IL-12. The underlying factors responsible for diminished IFN-γ production are not known, but may lie in the balance of stimulatory and inhibitory signals delivered to the IFN-γ secreting cells along with IL-12, or may relate more to the absence of memory T cells among cord blood MNC.
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Abbreviations
- CMI:
-
cell-mediated immunity
- IFN-γ:
-
interferon-γ
- MNC:
-
mononuclear cells
- NK:
-
natural killer
- CTL:
-
cytotoxic lymphocyte
- Th1:
-
T-helper type 1
- Th2:
-
T-helper type-2
- r:
-
recombinant
- RT:
-
reverse transcription
- PCR:
-
polymerase chain reaction
- GAPDH:
-
glyceraldehyde-3-phosphate dehydrogenase
- TNF-α:
-
tumor necrosis factor-α
References
Wilson CB, Penix L, Weaver WM, Melvin A, Lewis DB 1992 Ontogeny of T lymphocyte function in the neonate. Am J Reprod Immunol 28: 132–135
Wara DW, Barrett DJ 1979 Cell-mediated immunity in the newborn: clinical aspects. Pediatrics 64: 822–828
Kohl S 1989 The neonatal human's immune response to herpes simplex virus infection: a critical review. Pediatr Infect Dis J 8: 67–74
Kaplan J, Shope TC, Bollinger RO, Smith J 1982 Human newborns are deficient in natural killer activity. J Clin Immunol 2: 350–355
Trinchieri G 1993 Interleukin-12 and its role in the generation of Thl cells. Immunol Today 14: 335–338
Kobayashi M, Fitz L, Ryan M, Hewick RM, Clark SC, Chan S, Loudon R, Sherman F, Perussia B, Trinchieri G 1989 Identification and purification of natural killer cell stimulatory factor (NKSF), a cytokine with multiple biologic effects on human lymphocytes. J Exp Med 170: 827–845
Stern AS, Podlaski FJ, Hulmes JD, Pan YE, Quinn PM, Wolitzky AG, Familletti PC, Stremlo DL, Truitt T, Chizzonite R, Gately MK 1990 Purification to homogeneity and partial characterization of cytotoxic lymphocyte maturation factor from human B-lymphoblastoid cells. Proc Natl Acad Sci USA 87: 6808–6812
Chan SH, Perussia B, Gupta JW, Kobayashi M, Pospisil M, Young HA, Wolf SF, Young D, Clark SC, Trinchieri G 1991 Induction of interferon-γ production by natural killer cell stimulatory factor: characterization of the responder cells and synergy with other inducers. J Exp Med 173: 869–879
Perussia B, Chan SH, D'Andrea A, Tsuji K, Santoli D, Pospisil M, Young D, Wolf SF, Trinchieri G 1992 Natural killer (NK) cell stimulatory factor or IL-12 has differential effects on the proliferation of TCR-αβ+, TCR-γδ+ T lymphocytes, and NK cells. J Immunol 149: 3495–3502
Chehimi J, Starr SE, Frank I, Rengaraju M, Jackson SJ, Llanes C, Kobayashi M, Perussia B, Young D, Nickbarg E, Wolf SF, Trinchieri G 1992 Natural killer (NK) cell stimulatory factor increases the cytotoxic activity of NK cells from both healthy donors and human immunodeficiency virus-infected patients. J Exp Med 175: 789–796
Mehrotra PT, Wu D, Crim JA, Mostowski HS, Siegel JP 1993 Effects of IL-12 on the generation of cytotoxic activity in human CD8+ T lymphocytes. J Immunol 151: 2444–2452
Manetti R, Parronchi P, Giudizi MG, Piccinni M-P, Maggi E, Trinchieri G, Romagnani S 1993 Natural killer cell stimulatory factor(interleukin 12 [IL-12]) induces T helper type 1 (Th1)-specific immune responses and inhibits the development of IL-4-producing Th cells. J Exp Med 177: 1199–1204
Hsieh CS, Macatonia SE, Tripp CS, Wolf SF, O'Garra A, Murphy KM 1993 Development of Th1 CD4+ T cells through IL-12 produced by Listeria-induced macrophages. Science 260: 547–549
D'Andrea A, Rengaraju M, Valiante NM, Chehimi J, Kubin M, Aste M, Chan SH, Kobayashi M, Young D, Nickbarg E, Chizzonite R, Wolf SF, Trinchieri G 1992 Production of natural killer cell stimulatory factor(interleukin 12) by periphreal blood mononuclear cells. J Exp Med 176: 1387–1398
Trinchieri G, Rengaraju M, D'Andrea A, Valiante NM, Kubin M, Aste M, Chehimi J 1993 Producer cells of interleukin-12 [letter]. Immunol Today 14: 237–238
Macatonia SE, Hosken NA, Litton M, Vieira P, Hsieh CS, Culpepper JA, Wysocka M, Trinchieri G, Murphy KM, O'Garra A 1995 Dendritic cells produce IL-12 and direct the development of Th1 cells from naive CD4+ T cells. J Immunol 154: 5071–5079
Cassatella MA, Meda L, Gasperini S, D'Andrea A, Ma X, Trinchieri G 1995 Interleukin-12 production by human polymorphonuclear leukocytes. Eur J Immunol 25: 1–5
D'Andrea A, Aste AM, Valiante NM, Ma X, Kubin M, Trinchieri G 1993 Interleukin 10 (IL-10) inhibits human lymphocyte interferon-γ production by suppressing natural killer cell stimulatory factor/IL-12 synthesis in accessory cells. J Exp Med 178: 1041–1048
Kubin M, Kamoun M, Trinchieri G 1994 Interleukin 12 synergizes with B7/CD28 interaction in inducing efficient proliferation and cytokine production of human T cells. J Exp Med 180: 211–222
Chomczynski P, Sacchi N 1987 Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159
Samuels ML 1989 Statistics for the Life Sciences. Dellen Publishing Co., San Francisco, pp 294–322.
de Waal Malefyt R, Abrams J, Bennett B, Figdor CG, de Vries JE 1991 Interleukin 10 (IL-10) inhibits cytokine synthesis by human monocytes: an autoregulatory role of IL-10 produced by monocytes. J Exp Med 174: 1209–1220
Hunter CA, Subauste CS, Van Cleave VH, Remington JS 1994 Production of γ interferon by natural killer cells fromToxoplasma gondii- infected SCID mice: regulation by interleukin-10, interleukin-12, and tumor necrosis factor alpha. Infect Immun 62: 2818–2824
Farrar MA, Schreiber RD 1993 The molecular cell biology of interferon-γ and its receptor. Annu Rev Immunol 11: 571–611
Bryson YJ, Winter HS, Gard SE, Fischer TJ, Stiehm ER 1980 Deficiency of immune interferon production by leukocytes of normal newborns. Cell Immunol 55: 191–200
Wilson CB, Westall J, Johnston L, Lewis DB, Dower SK, Alpert AR 1986 Decreased production of interferon-γ by human neonatal cells. Intrinsic and regulatory deficiencies. J Clin Invest 77: 860–867
Kruse A, Rink L, Rutenfranz I, Kolanczyk B, Kirchner H 1992 Interferon and lymphokine production by human placental and cord blood cells. J Interferon Res 12: 113–117
Burchett SK, Corey L, Mohan KM, Westall J, Ashley R, Wilson CB 1992 Diminished interferon-γ and lymphocyte proliferation in neonatal and postpartum primary herpes simplex virus infection. J Infect Dis 165: 813–818
Lewis DB, Larsen A, Wilson CB 1986 Reduced interferon-γ mRNA levels in human neonates. Evidence for an intrinsic T cell deficiency independent of other genes involved in T cell activation. J Exp Med 163: 1018–1023
Lewis DB, Yu CC, Meyer J, English BK, Kahn SJ, Wilson CB 1991 Cellular and molecular mechanisms for reduced interleukin-4 and interferon-γ production by neonatal T cells. J Clin Invest 87: 194–202
Wakasugi N, Virelizier JL 1985 Defective IFN-γ production in the human neonate. I. Dysregulation rather than intrinsic abnormality. J Immunol 134: 167–171
Wakasugi N, Virelizier JL, Arenzana-Seisdedos F, Rothhut B, Huerta JM, Russo-Marie F, Fiers W 1985 Defective IFN-γ production in the human neonate. II. Role of increased sensitivity to the suppressive effects of prostaglandin E. J Immunol 134: 172–176
Lau AS, Sigaroudinia M, Yeung MC, Kohl S 1996 Interleukin-12 induces interferon-γ expression and natural killer cytotoxicity in cord blood mononuclear cells. Pediatr Res 39: 150–155
Fiorentino DF, Zlotnik A, Vieira P, Mosmann TR, Howard M, Moore KW, O'Garra A 1991 IL-10 acts on the antigen-presenting cell to inhibit cytokine production by Th1 cells. J Immunol 146: 3444–3451
Tufano MA, Cpiollaro de l'Ero G, Ianniello R, Galdiero M, Galdiero F 1991 Protein A and other surface components of Staphylococcus aureus stimulate production of IL-1α, IL-4, IL-6, TNF and IFN-γ. Eur Cytokine Netw 2: 361–366.
Ding L, Linsley PS, Huang LY, Germain RN, Shevach EM 1993 IL-10 inhibits macrophage costimulatory activity by selectively inhibiting the up-regulation of B7 expression. J Immunol 151: 1224–1234
Chomarat P, Rissoan MC, Banchereau J, Miossec P 1993 Interferon-γ inhibits interleukin 10 production by monocytes. J Exp Med 177: 523–527
Chheda S, Palkowetz KH, Garofalo R, Rassin DK, Goldman AS 1996 Decreased interleukin-10 production by neonatal monocytes and T cells: relationship to decreased production and expression of tumor necrosis factor-α and its receptors. Pediatr Res 40: 475–483
Ling P, Gately MK, Gubler U, Stern AS, Lin P, Hollfelder K, Su C, Pan YC, Hakimi J 1995 Human IL-12 p40 homodimer binds to the IL-12 receptor but does not mediate biologic activity. J Immunol 154: 116–127
Lin H, Mosmann TR, Guilbert L, Tuntipopipat S, Wegmann TG 1993 Synthesis of T helper 2-type cytokines at the maternal-fetal interface. J Immunol 151: 4562–4573
Frenkel L, Bryson YJ 1987 Ontogeny of phytohemagglutinin-induced gamma interferon by leukocytes of healthy infants and children: evidence for decreased production in infants younger than 2 months of age. J Pediatr 111: 97–100
Jung T, Schauer U, Heusser C, Neumann C, Rieger C 1993 Detection of intracellular cytokines by flow cytometry. J Immunol Methods 159: 197–207
Wu C-Y, Demeure C, Kiniwa M, Gately M, Delespesse G 1993 IL-12 induces the production of IFN-γ by neonatal human CD4 T cells. J Immunol 151: 1938–1949
Brugnoni D, Airo P, Graf D, Marconi M, Lebowitz M, Plebani A, Giliani S, Malacarne F, Cattaneo R, Ugazio AG, Albertini A, Kroczek RA, Notarangelo LD 1994 Ineffective expression of CD40 ligand on cord blood T cells may contribute to poor immunoglobulin production in the newborn. Eur J Immunol 24: 1919–1924
Nonoyama S, Penix LA, Edwards CP, Lewis DB, Ito S, Aruffo A, Wilson CB, Ochs HD 1995 Diminished expression of CD40 ligand by activated neonatal T cells. J Clin Invest 95: 66–75
Armitage RJ, Tough TW, Macduff BM, Fanslow WC, Spriggs MK, Ramsdell F, Alderson MR 1993 CD40 ligand is a T cell growth factor. Eur J Immunol 23: 2326–2331
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Supported in part by National Institutes of Health Grants AI32384 and AI07395-03.1 Current address: Immunex Research & Development Corp., Seattle, WA 98101.
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Scott, M., Kubin, M. & Kohl, S. High Level Interleukin-12 Production, but Diminished Interferon-γ Production, by Cord Blood Mononuclear Cells. Pediatr Res 41, 547–553 (1997). https://doi.org/10.1203/00006450-199704000-00015
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DOI: https://doi.org/10.1203/00006450-199704000-00015
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