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. 1998 Nov;43(5):620–628. doi: 10.1136/gut.43.5.620

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

G Monteleone 1, T Parrello 1, F Luzza 1, F Pallone 1
PMCID: PMC1727325  PMID: 9824341

Abstract

Background/Aim—Interleukin (IL) 12 is involved in the mucosal response during intestinal inflammation but its role is not fully understood. The response of human lamina propria T lymphocytes (T-LPL) to IL-12 in terms of interferon γ (IFN-γ) release and proliferation was investigated, exploring whether IL-15 and IL-7 cooperate with IL-12. The role of accessory molecules (CD2 and CD28) was also investigated. 
Methods—Unstimulated and phytohaemagglutinin preactivated T-LPL cultures were incubated with or without the initial addition of cytokines, anti-CD2 or anti-CD28 antibodies. IFN-γ mRNA was detected by reverse transcriptase polymerase chain reaction, and protein secretion was measured by enzyme linked immunosorbent assay (ELISA). 
Results—IFN-γ mRNA was induced in T-LPLs by IL-12 and IL-15 but not IL-7, whereas IFN-γ was measured only in IL-12 stimulated T-LPL cultures. IL-12 induced IFN-γ release was not abrogated by neutralising anti-IL-2 antibody or by cyclosporin A. IL-12 synergised with either anti-CD2 or anti-CD28 antibodies in inducing IFN-γ synthesis. In preactivated T-LPLs, IL-7 enhanced IFN-γ release induced by both IL-12 and anti-CD2, whereas IL-15 potentiated only IL-12 induced IFN-γ synthesis. IL-12 did not induce proliferation of either unstimulated or preactivated T-LPLs and it did not enhance the CD2/CD28 stimulated T-LPL proliferative response. No transcript for IL-12 receptor β1 subunit was detected in freshly isolated and activated T-LPLs whereas the β2 subunit mRNA was consistently found in T-LPL samples. 
Conclusions—IL-12 induces human T-LPLs to produce and release IFN-γ, and IL-15 and IL-7 cooperate with IL-12 in expanding the IFN-γ mucosal response. 



Keywords: interleukin 12; interleukin 7; interleukin 15; interferon γ; intestinal inflammation

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Figure 1 .

Figure 1

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Interferon γ (IFN-γ) release in three day cultures of both interleukin (IL) 12- and IL-12 (1000 pg/ml final concentration) plus anti-IL-12 antibody-stimulated lamina propria T lymphocytes. Each point on the curve represents the mean of all experiments; vertical bars indicate 1 SD. At each time interval, IL-12 induced IFN-γ release in a dose dependent manner (p = 0.01). 


Figure 2 .

Figure 2

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Interferon γ (IFN-γ) release in both lamina propria T lymphocytes (T-LPLs) and phytohaemagglutinin (PHA) preactivated T-LPLs after 24 hours of culture. Both T-LPLs and PHA preactivated T-LPLs were incubated with graded doses of interleukin (IL) 12 in the presence or absence of anti-CD2 antibody. Each point on the curve represents the mean of six representative experiments; vertical bars indicate 1 SD. Values of IFN-γ measured after co-stimulation with IL-12 and anti-CD2 antibody were significantly higher than observed after stimulation with IL-12 (p<0.0001) or anti-CD2 antibody alone (p<0.0001). 


Figure 3 .

Figure 3

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Interferon γ (IFN-γ) release by phytohaemagglutinin preactivated lamina propria T lymphocytes (T-LPLs) after 24 hours of culture. Cells were cultured with interleukin (IL) 12 (100 pg/ml final concentration) or anti-CD28 (1 µg/ml final concentration) or anti-CD2 (1:1000 final dilution) antibody in the presence or absence of IL-7 (10 ng/ml final concentration) or IL-15 (10 ng/ml final concentration). Data are the mean of six representative experiments; horizontal bars indicate 1 SD. The co-stimulation with IL-7 significantly enhanced T-LPL IFN-γ release induced by IL-12 (p<0.0001) or anti-CD28 (p<0.0001) or CD2 (p<0.0001) antibody. IL-15 potentiated IL-12 stimulated T-LPL IFN-γ release (p<0.0001) but not that induced by anti-CD28 or CD2 antibody. 


Figure 4 .

Figure 4

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Agarose gel stained with ethidium bromide showing RT-PCR products for IL-12Rβ1 (557 bp; A) (after 35 cycles), IL-12Rβ2 (298 bp; B) (after 35 cycles), and glyceraldehyde-3-phosphate dehydrogenase (368 bp; C) (after 22 cycles) in lamina propria T lymphocytes (T-LPLs) and autologous peripheral blood T lymphocytes (T-PBLs) cultured with or without stimulation. Lane 1, 123 bp ladder; lane 2, unstimulated T-LPLs; lane 3, PHA (1µg/ml) stimulated T-LPLs; lane 4, anti-CD2/anti-CD28 antibody stimulated T-LPLs; lane 5, unstimulated T-PBLs; lane 6, PHA (1µg/ml) stimulated T-PBLs; lane 7, anti-CD2/anti-CD28 antibody stimulated T-PBLs. 


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