Enhancing lamina propria Th1 cell responses with interleukin 12 produces severe tissue injury

doi:10.1016/S0016-5085(99)70391-4

Gastroenterology

Volume 117, Issue 5, November 1999, Pages 1069-1077

https://doi.org/10.1016/S0016-5085(99)70391-4 Get rights and content

Abstract

Background & Aims: Interleukin (IL)-12 is believed to modulate local T-cell response in human colitis. A direct functional relationship between IL-12 and tissue injury in human intestine has not been reported. The aim of this study was to examine changes that take place in explant cultures of human fetal gut after stimulation of T cells with anti-CD3 in the presence of exogenous IL-12/IL-18. Methods: T cells in explants of fetal gut were activated with anti-CD3 antibody and/or IL-12 or IL-18. Mucosal pathology was determined by immunohistochemistry. Quantitative reverse-transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay were used to determine cytokine synthesis, and the production of matrix metalloproteinases was analyzed by RT-PCR and Western blotting. Results: Activation of T cells in explants with anti-CD3 antibody elicited very little interferon (IFN)-γ and tumor necrosis factor (TNF)-α production and no tissue injury. Addition of graded doses of IL-12 with anti-CD3 resulted in a significant increase in both IFN-γ and TNF-α. This change was associated with a massive increase in stromelysin-1 expression and severe tissue injury, which was inhibitable by a stromelysin-1 inhibitor. Costimulation of explants with anti-CD3 and IL-18 induced only IFN-γ and no tissue injury. Conclusions: IL-12 can convert a physiological T-cell signal into a strong signal with the downstream effect of elevating tissue stromelysin-1 concentration and mucosal degradation.

GASTROENTEROLOGY 1999;117:1069-1077

Section snippets

Organ culture of human fetal gut

The study received ethical approval from the Hackney and District Health Authority (London, England). Human fetal intestinal tissue was obtained within 2 hours of surgical termination from the Homerton Hospital (London, England). All specimens were aged between 15 and 16 weeks of gestation. Fetal gut explants were cultured in the presence of recombinant human IL-12 (final concentration ranging from 0.5 to 10 ng/mL) and/or IL-18 (100 ng/mL; both purchased from R&D Systems, Minneapolis, MN)

IL-12Rβ2 and IL-1Rrp but not IL-12/p40 are expressed in human fetal gut

To examine whether IL-12 was expressed in human fetal gut, RNA was extracted from the small intestine and tested for IL-12/P40 mRNA by RT-PCR. We have shown that IL-12/P40 is expressed in CD.¹⁴ RNA extracted from mucosal tissue of a patient with CD was used as positive control. As shown in Figure 1, no IL-12/P40 transcripts were detected in freshly isolated explants.

. Agarose gel stained with ethidium bromide showing RT-PCR products for IL-12 p40, IL-12Rβ2, IL-1Rrp, and GAPDH in freshly obtained

Discussion

In this study we show that addition of IL-12 with soluble CD3 antibody to explant cultures of fetal gut results in activation of effector cells capable of inducing tissue injury. The effect of IL-12 was dose dependent and could be neutralized by an IL-12 antibody. To the best of our knowledge, this is the first mechanistic study showing a functional relationship between IL-12 and human intestinal mucosal injury.

Both in vivo and in vitro studies have documented the ability of IL-12 to induce Th1

Acknowledgements

The authors thank Prof. Ian Sanderson for helpful comments and Dr. G. S. Schultz (University of Florida) for matrix metalloproteinase synthetic RNA templates.

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^☆: Address requests for reprints to: Sylvia L. F. Pender, Ph.D., Department of Paediatric Gastroenterology, St. Bartholomew's and the Royal London School of Medicine and Dentistry, Suite 31, 3rd Floor, Dominion House, 59 Bartholomew's Close, London, EC1A 7BE, England. e-mail: [email protected]; fax: (44) 207-6005901.

^☆☆: Supported by the European Union grant ERBFMRXCT9, the special trustee of St. Bartholomew's Hospital, and Crohn's in Childhood Research Association.

View full text

Alimentary TractEnhancing lamina propria Th1 cell responses with interleukin 12 produces severe tissue injury☆,☆☆

Abstract

Section snippets

Organ culture of human fetal gut

IL-12Rβ2 and IL-1Rrp but not IL-12/p40 are expressed in human fetal gut

Discussion

Acknowledgements

Immunity

Gastroenterology

Gastroenterology

Gastroenterology

J Biol Chem

Gastroenterology

Cloning of cDNA for natural killer cell stimulatory factor, a heterodimeric cytokine with multiple biologic effects on T and natural killer cells

J Immunol

Interleukin-12: a cytokine produced by antigen-presenting cells with immunoregulatory functions in the generation of T-helper cells type 1 and cytotoxic lymphocytes

Blood

Interleukin 12 acts directly on CD4+ T cells to enhance priming for interferon γ production and diminishes interleukin 4 inhibition of such priming

Proc Natl Acad Sci USA

Interleukin 12 aministration induces T helper type 1 cells and accelerates autoimmune diabetes in NOD mice

J Exp Med

Cloning of a new cytokine that induces IFN-γ production by T cells

Nature

IFN-γ–inducing factor (IGIF) is a costimulatory factor on the activation of Th1 but not Th2 cells and exerts its effect independently of IL-12

J Immunol

A mechanism underlying synergy between IL-12 and IFN-gamma–inducing factor in enhanced production of IFN-gamma

J Immunol

IL-18 and CD28 use distinct molecular mechanisms to enhance NK cell production of IL-12–induced IFN-γ

J Immunol

Interleukin-2 and interferon-γ secreting T cells in normal and diseased human intestinal mucosa

Immunology

Spontaneous release of interferon gamma by intestinal lamina propria lymphocytes in Crohn's disease. Kinetics of in vitro response to interferon gamma inducers

Gut

Disparate CD4+ lamina propria (LP) lymphokine secretion profiles in inflammatory bowel disease. Crohn's disease LP cells manifest increased secretion of IFN-γ, whereas ulcerative colitis LP cells manifest increased secretion of IL-5

J Immunol

Type 1 T-helper cell predominance and interleukin-12 expression in the gut of patients with Crohn's disease

Am J Pathol

Interleukin-12 expression is focally enhanced in the gastric mucosa of pediatric patients with Crohn' s disease

Am J Pathol

Response of human intestinal lamina propria T lymphocytes to interleukin 12: additive effects of interleukin 15 and 7

Gut

Antibodies to interleukin 12 abrogate established experimental colitis in mice

J Exp Med

Alimentary Tract
Enhancing lamina propria Th1 cell responses with interleukin 12 produces severe tissue injury☆,☆☆

Interleukin 12 acts directly on CD4⁺ T cells to enhance priming for interferon γ production and diminishes interleukin 4 inhibition of such priming

Disparate CD4⁺ lamina propria (LP) lymphokine secretion profiles in inflammatory bowel disease. Crohn's disease LP cells manifest increased secretion of IFN-γ, whereas ulcerative colitis LP cells manifest increased secretion of IL-5