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Selective expression of an interleukin-12 receptor component by human T helper 1 cells.

Rogge L, Barberis-Maino L, Biffi M, Passini N, Presky DH, Gubler U, Sinigaglia F - J. Exp. Med. (1997)

Bottom Line: Here we show that the IL-12 receptor (IL-12R) beta 2 subunit, a recently cloned binding and signal transducing component of the IL-12R, is expressed on human Th1 but not Th2 clones and is induced during differentiation of human naive cells along the Th1 but not the Th2 pathway.IL-12 and type I but not type II interferons induce expression of the IL-12R beta 2 chain during in vitro T cell differentiation after antigen receptor triggering.The selective expression and regulation of the IL-12R beta 2 subunit may help to understand the basis of Th1/Th2 differentiation and may provide therapeutic options for altering the Th1/Th2 balance in several immuno-pathological conditions such as autoimmune diseases and allergies.

View Article: PubMed Central - PubMed

Affiliation: Roche Milano Ricerche, Italy.

ABSTRACT
Interleukin-12 (IL-12), a heterodimeric cytokine produced by activated monocytes and dendritic cells, plays a crucial role in regulating interferon (IFN)-gamma production and in the generation of IFN-gamma-producing T helper 1 (Th1) cells. Here we show that the IL-12 receptor (IL-12R) beta 2 subunit, a recently cloned binding and signal transducing component of the IL-12R, is expressed on human Th1 but not Th2 clones and is induced during differentiation of human naive cells along the Th1 but not the Th2 pathway. IL-12 and type I but not type II interferons induce expression of the IL-12R beta 2 chain during in vitro T cell differentiation after antigen receptor triggering. The selective expression and regulation of the IL-12R beta 2 subunit may help to understand the basis of Th1/Th2 differentiation and may provide therapeutic options for altering the Th1/Th2 balance in several immuno-pathological conditions such as autoimmune diseases and allergies.

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Transcripts encoding IL-12 receptor subunits in human Th1  and Th2 cells. (A) IL-12R β2 transcripts are selectively expressed in human Th1 cells. Th1 and Th2 lines generated from cord blood lymphocytes were harvested on the indicated time after priming. In lanes 9 and  10, cells were washed on day 19 and incubated for 48 h in medium containing 2 ng/ml IL-12 before RNA extraction on day 21. Transcripts encoding the IL-12R β1 subunit (top), the IL-12R β2 subunit (middle) and  an 18S RNA gene fragment as loading control (bottom) were quantitated  with ribonuclease protection assays as described in Materials and Methods.  (B) Expression of IL-12R β1 and β2 mRNAs in human Th1 (CB13) and  Th2 (CB21) T cell clones. Th1 and Th2 cells were incubated on day 11  after restimulation for 48 h with or without 2 ng/ml IL-12, followed by  RNA extraction on day 13. Quantitation of the IL-12R β1 and β2 transcripts in IL-12 treated and untreated Th1 and Th2 cells was performed  by RNase protection assays.
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Figure 3: Transcripts encoding IL-12 receptor subunits in human Th1 and Th2 cells. (A) IL-12R β2 transcripts are selectively expressed in human Th1 cells. Th1 and Th2 lines generated from cord blood lymphocytes were harvested on the indicated time after priming. In lanes 9 and 10, cells were washed on day 19 and incubated for 48 h in medium containing 2 ng/ml IL-12 before RNA extraction on day 21. Transcripts encoding the IL-12R β1 subunit (top), the IL-12R β2 subunit (middle) and an 18S RNA gene fragment as loading control (bottom) were quantitated with ribonuclease protection assays as described in Materials and Methods. (B) Expression of IL-12R β1 and β2 mRNAs in human Th1 (CB13) and Th2 (CB21) T cell clones. Th1 and Th2 cells were incubated on day 11 after restimulation for 48 h with or without 2 ng/ml IL-12, followed by RNA extraction on day 13. Quantitation of the IL-12R β1 and β2 transcripts in IL-12 treated and untreated Th1 and Th2 cells was performed by RNase protection assays.

Mentions: We next examined whether the defect in IL-12 signaling observed in Th2 cells is due to a differential expression of IL-12 receptors in the two cell types. To date, two IL-12R subunits, termed β1 and β2, have been identified. IL-12R β1 or β2, independently expressed in COS cells, bind human IL-12 with low affinity (KD = 3–5 nM), whereas coexpression of IL-12R β1 and β2 in COS cells gives rise to high affinity binding sites (KD = 50 pm) (25, 26). These studies have also shown that the IL-12R β2 is the signaling component of the IL-12R, in agreement with the fact that the IL-12R β2, in contrast to the IL-12R β1, contains tyrosine residues in its cytoplasmic domain (25). We analyzed the mRNA expression levels for the IL-12R β1 and β2 subunits by ribonuclease protection assays in both Th1 and Th2 lines at different time points after priming and in Th1 and Th2 clones at day 13 after restimulation. Whereas IL-12R β1 transcripts were expressed in similar amounts in both Th1 and Th2 cells (Fig. 3, A and B), the transcripts coding for the IL-12R β2 subunit were selectively expressed in established Th1, but not Th2 lines (Fig. 3 A) and clones (Fig. 3 B). Maximum expression in Th1 cells was seen between day 3 and day 8, and declined thereafter. Th2 cells, in contrast, expressed little IL-12R β2 transcripts on day 3, and none were detected after day 8 (Fig. 3 A). A key question is whether cytokine signaling is required for IL-12R induction or whether stimulation via the TCR/CD3 complex is sufficient to induce IL-12R in naive T cells. IL-12R transcripts were therefore analyzed in purified CD45RA+ T cells before and after stimulation with plate-bound anti-CD3 mAb. IL-12R transcripts were not found in naive T cells (Fig. 4, lane 2), but could be detected as early as 24 h after T cell activation (Fig. 4, lane 9). Priming of naive T cells in the presence of IL-12 resulted in enhanced expression of IL-12R β2 transcripts in this population (Fig. 4, lane 13); priming of cells in the presence of IL-4, in contrast, resulted in very low levels of IL-12R β2 transcripts (Fig. 4, lane 14). Thus, antigen receptor triggering is sufficient to make naive T cells susceptible to IL-12 signaling.


Selective expression of an interleukin-12 receptor component by human T helper 1 cells.

Rogge L, Barberis-Maino L, Biffi M, Passini N, Presky DH, Gubler U, Sinigaglia F - J. Exp. Med. (1997)

Transcripts encoding IL-12 receptor subunits in human Th1  and Th2 cells. (A) IL-12R β2 transcripts are selectively expressed in human Th1 cells. Th1 and Th2 lines generated from cord blood lymphocytes were harvested on the indicated time after priming. In lanes 9 and  10, cells were washed on day 19 and incubated for 48 h in medium containing 2 ng/ml IL-12 before RNA extraction on day 21. Transcripts encoding the IL-12R β1 subunit (top), the IL-12R β2 subunit (middle) and  an 18S RNA gene fragment as loading control (bottom) were quantitated  with ribonuclease protection assays as described in Materials and Methods.  (B) Expression of IL-12R β1 and β2 mRNAs in human Th1 (CB13) and  Th2 (CB21) T cell clones. Th1 and Th2 cells were incubated on day 11  after restimulation for 48 h with or without 2 ng/ml IL-12, followed by  RNA extraction on day 13. Quantitation of the IL-12R β1 and β2 transcripts in IL-12 treated and untreated Th1 and Th2 cells was performed  by RNase protection assays.
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Figure 3: Transcripts encoding IL-12 receptor subunits in human Th1 and Th2 cells. (A) IL-12R β2 transcripts are selectively expressed in human Th1 cells. Th1 and Th2 lines generated from cord blood lymphocytes were harvested on the indicated time after priming. In lanes 9 and 10, cells were washed on day 19 and incubated for 48 h in medium containing 2 ng/ml IL-12 before RNA extraction on day 21. Transcripts encoding the IL-12R β1 subunit (top), the IL-12R β2 subunit (middle) and an 18S RNA gene fragment as loading control (bottom) were quantitated with ribonuclease protection assays as described in Materials and Methods. (B) Expression of IL-12R β1 and β2 mRNAs in human Th1 (CB13) and Th2 (CB21) T cell clones. Th1 and Th2 cells were incubated on day 11 after restimulation for 48 h with or without 2 ng/ml IL-12, followed by RNA extraction on day 13. Quantitation of the IL-12R β1 and β2 transcripts in IL-12 treated and untreated Th1 and Th2 cells was performed by RNase protection assays.
Mentions: We next examined whether the defect in IL-12 signaling observed in Th2 cells is due to a differential expression of IL-12 receptors in the two cell types. To date, two IL-12R subunits, termed β1 and β2, have been identified. IL-12R β1 or β2, independently expressed in COS cells, bind human IL-12 with low affinity (KD = 3–5 nM), whereas coexpression of IL-12R β1 and β2 in COS cells gives rise to high affinity binding sites (KD = 50 pm) (25, 26). These studies have also shown that the IL-12R β2 is the signaling component of the IL-12R, in agreement with the fact that the IL-12R β2, in contrast to the IL-12R β1, contains tyrosine residues in its cytoplasmic domain (25). We analyzed the mRNA expression levels for the IL-12R β1 and β2 subunits by ribonuclease protection assays in both Th1 and Th2 lines at different time points after priming and in Th1 and Th2 clones at day 13 after restimulation. Whereas IL-12R β1 transcripts were expressed in similar amounts in both Th1 and Th2 cells (Fig. 3, A and B), the transcripts coding for the IL-12R β2 subunit were selectively expressed in established Th1, but not Th2 lines (Fig. 3 A) and clones (Fig. 3 B). Maximum expression in Th1 cells was seen between day 3 and day 8, and declined thereafter. Th2 cells, in contrast, expressed little IL-12R β2 transcripts on day 3, and none were detected after day 8 (Fig. 3 A). A key question is whether cytokine signaling is required for IL-12R induction or whether stimulation via the TCR/CD3 complex is sufficient to induce IL-12R in naive T cells. IL-12R transcripts were therefore analyzed in purified CD45RA+ T cells before and after stimulation with plate-bound anti-CD3 mAb. IL-12R transcripts were not found in naive T cells (Fig. 4, lane 2), but could be detected as early as 24 h after T cell activation (Fig. 4, lane 9). Priming of naive T cells in the presence of IL-12 resulted in enhanced expression of IL-12R β2 transcripts in this population (Fig. 4, lane 13); priming of cells in the presence of IL-4, in contrast, resulted in very low levels of IL-12R β2 transcripts (Fig. 4, lane 14). Thus, antigen receptor triggering is sufficient to make naive T cells susceptible to IL-12 signaling.

Bottom Line: Here we show that the IL-12 receptor (IL-12R) beta 2 subunit, a recently cloned binding and signal transducing component of the IL-12R, is expressed on human Th1 but not Th2 clones and is induced during differentiation of human naive cells along the Th1 but not the Th2 pathway.IL-12 and type I but not type II interferons induce expression of the IL-12R beta 2 chain during in vitro T cell differentiation after antigen receptor triggering.The selective expression and regulation of the IL-12R beta 2 subunit may help to understand the basis of Th1/Th2 differentiation and may provide therapeutic options for altering the Th1/Th2 balance in several immuno-pathological conditions such as autoimmune diseases and allergies.

View Article: PubMed Central - PubMed

Affiliation: Roche Milano Ricerche, Italy.

ABSTRACT
Interleukin-12 (IL-12), a heterodimeric cytokine produced by activated monocytes and dendritic cells, plays a crucial role in regulating interferon (IFN)-gamma production and in the generation of IFN-gamma-producing T helper 1 (Th1) cells. Here we show that the IL-12 receptor (IL-12R) beta 2 subunit, a recently cloned binding and signal transducing component of the IL-12R, is expressed on human Th1 but not Th2 clones and is induced during differentiation of human naive cells along the Th1 but not the Th2 pathway. IL-12 and type I but not type II interferons induce expression of the IL-12R beta 2 chain during in vitro T cell differentiation after antigen receptor triggering. The selective expression and regulation of the IL-12R beta 2 subunit may help to understand the basis of Th1/Th2 differentiation and may provide therapeutic options for altering the Th1/Th2 balance in several immuno-pathological conditions such as autoimmune diseases and allergies.

Show MeSH
Related in: MedlinePlus