Interleukin 12 and interleukin 4 control T cell adhesion to endothelial selectins through opposite effects on alpha1, 3-fucosyltransferase VII gene expression - PubMed (original) (raw)

Interleukin 12 and interleukin 4 control T cell adhesion to endothelial selectins through opposite effects on alpha1, 3-fucosyltransferase VII gene expression

A J Wagers et al. J Exp Med. 1998.

Abstract

The alpha1,3-fucosyltransferase, FucT-VII, is crucial for the formation of ligands for all three selectins, and its expression regulates the synthesis of these ligands. Short-term polarized T helper (Th)1, but not Th2 or naive CD4(+) T cells, can home to sites of inflammation, but the molecular basis for this difference has remained unclear. Here we show that naive CD4(+) T cells do not express FucT-VII and fail to bind vascular selectins. We also show that when CD4(+) T cells are activated in the presence of the Th1 polarizing cytokine interleukin (IL)-12, levels of FucT-VII mRNA and binding to E- and P-selectin are significantly augmented. In contrast, activation of CD4(+) T cells in the presence of IL-4, a Th2 polarizing cytokine, inhibited FucT-VII expression and binding to vascular selectins. T cell activation upregulated expression of the Core2 transferase, C2GnT, equivalently regardless of the presence or absence of polarizing cytokines. These data indicate that the selective ability of Th1 cells, as opposed to Th2 cells or naive CD4(+) T cells, to recognize vascular selectins and home to sites of inflammation is controlled principally by the expression of a single gene, FucT-VII.

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Figures

Figure 1

Figure 1

Naive CD4+ T cells do not express FucT-VII and do not bind to vascular selectins. (A) Expression of FucT-VII and C2GnT mRNA by unfractionated, naive, and memory CD4+ T lymphocytes was determined by semiquantitative RT-PCR analysis, as described in Materials and Methods. RT indicates reverse transcriptase; + indicates reactions in which RT was included; − indicates control reactions in which RT was omitted. PGK1 is a housekeeping gene used as an internal control for the RT reaction. (B) Expression of HECA-452 mAb epitope by purified naive and memory CD4+ T lymphocytes was determined by FACS® analysis. Numbers in the upper right corner of each histogram indicate the percent HECA-452 positive cells in each population. (C) Binding of unfractionated, naive, and memory CD4+ T lymphocytes to COS cells transiently transfected with E-selectin (solid bars) or P-selectin (gray bars). Data are presented as mean numbers of lymphocytes bound per COS cell ± SEM.

Figure 2

Figure 2

Effect of Th1 or Th2 polarizing conditions on expression of FucT-VII and C2GnT mRNA. RNA was isolated from fresh CD4+ T lymphocytes or after activation with PHA and culture with IL-2 alone, IL-2 + IL-4, or IL-2 + IL-12. (A) Expression of FucT-VII and C2GnT mRNA was determined by semiquantitative RT-PCR, as in Fig. 1. (B) A more detailed analysis of FucT-VII and C2GnT expression was carried out using twofold serial dilutions of input cDNA, using the same amount of cDNA in the first (left) lane as in A. Amounts of input cDNA decrease to the left, as indicated above the blots. The lanes on the right (−RT) are the negative controls in which the RT is omitted from the RT reaction.

Figure 2

Figure 2

Effect of Th1 or Th2 polarizing conditions on expression of FucT-VII and C2GnT mRNA. RNA was isolated from fresh CD4+ T lymphocytes or after activation with PHA and culture with IL-2 alone, IL-2 + IL-4, or IL-2 + IL-12. (A) Expression of FucT-VII and C2GnT mRNA was determined by semiquantitative RT-PCR, as in Fig. 1. (B) A more detailed analysis of FucT-VII and C2GnT expression was carried out using twofold serial dilutions of input cDNA, using the same amount of cDNA in the first (left) lane as in A. Amounts of input cDNA decrease to the left, as indicated above the blots. The lanes on the right (−RT) are the negative controls in which the RT is omitted from the RT reaction.

Figure 3

Figure 3

HECA-452 reactivity of CD4+ T cells in response to varying culture conditions. HECA-452 staining of FucT-VII reporter epitopes (5) was used to assess FucT-VII activity. Naive CD4+ T cells (A) or unfractionated CD4+ T cells (B–D) were activated with either anti-CD3/anti-CD28 (A and B), PHA-P (C), or TSST-1 (D). In all panels, squares denote cultures containing IL-2 alone, diamonds denote cultures containing IL-2 + IL-4, and circles denote cultures containing IL-2 + IL-12. Data are presented as percent HECA-452 positive lymphocytes for each culture condition at the indicated time point.

Figure 4

Figure 4

Binding of Th1- or Th2-type cells to E- and P-selectin. (A and B) Expression of selectin ligands by anti-CD3/anti-CD28 activated CD4+ T lymphocytes was determined by flow cytometry using RIgM chimera proteins. Data are presented as percentage of cells staining with E-RIgM (A) or P-RIgM (B) chimera for each culture condition at the indicated time point. Squares denote cultures containing IL-2 alone, diamonds denote cultures containing IL-2 + IL-4, and circles denote cultures containing IL-2 + IL-12. (C and D) Binding of activated CD4+ T cells to vascular selectins was assessed in in vitro rolling assays at 1.7 dyn/ cm2. Cells were activated with anti-CD3/anti-CD28 and analyzed 2 d after the second activation. HECA-452 reactivity of the analyzed cells was as follows: IL-2 only, 32.7% positive; IL-2 + IL-4, 9.9% positive; IL-2 + IL-12, 48.3% positive. Data are expressed as the mean number of rolling cells/field/min ± SD for at least 10 different fields of view. (C) Rolling interactions of CD4+ T lymphocytes on CHO/E-selectin monolayers. (D) Rolling interactions of CD4+ T lymphocytes on CHO/P-selectin monolayers. (*, **) For each selectin, differences in rolling among unpolarized, Th1-, and Th2-type CD4+ T cell groups were statistically significant (P < 0.05).

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