Human memory T cell differentiation into Th2-like effector cells is dependent on IL-4 and CD28 stimulation and inhibited by TCR ligation

Abstract

Freshly isolated memory T cells primarily produced IL-2 and small amounts of IL-4 and IFN-+ after stimulation in vitro. Priming for 5 days in vitro with anti-CD28 monoclonal antibodies (mAb) alone markedly increased production of IL-4. In comparison to fresh cells, the increase in the amount of IL-4 secreted reflected a marked increase in the number of IL-4-producing cells. Stimulation with immobilized anti-CD3 mAb during priming limited subsequent IL-4 production. By contrast, IFN-+ production from in vitro primed memory T cells was directly correlated to the concentration of priming anti-CD3 mAb. IL-2 production by all restimulated cells was decreased. The differentiation of IL-4-producing cells could be blocked by antibody to IL-4 and enhanced by the addition of recombinant IL-4 as well as antibody to IFN-+ . Of note, the IL-4-producing effector cells induced from in vitro priming derived from the early CD27 pos memory T cell subset, whereas the small CD27 neg differentiated memory subset produced IL-4 without in vitro priming. The results indicate that memory T cells can be directed to differentiate into IL-4-producing effector cells by stimulation via CD28 and IL-4, whereas increasing engagement of the TCR limits Th2 memory cell differentiation.

Differential Antigen Sensitivity and Costimulatory Requirements in Human Th1 and Th2 Antigen-Specific CD4+ Cells with Similar TCR Avidity

“…In studies of human T cells activated with anti-CD3, Th2 effector cells were dependent on CD28 ligation but were inhibited by increased signal via the TCR, again suggesting that lower binding potential of the MHC-peptide-TCR complex would favor Th2 cell development (14). To study the Ag-specific human CD4 ϩ T cell response, we have examined the activation requirements and affinity of memory T cells which respond to the same peptide-MHC complex but are committed to either Th1 or Th2 lineage.…”

Restriction of De Novo Pyrimidine Biosynthesis Inhibits Th1 Cell Activation and Promotes Th2 Cell Differentiation

“…Cells were washed with PBS and incubated with SRBC (30). The rosette-positive cells were further purified by negative selection using saturating amounts of mAbs against CD8, CD16, CD19, CD56, HLA-DR, and CD45RA (for the purification of memory CD4 T cells) or CD45RO (for the purification of naive CD4 T cells), followed by panning on plastic petri dishes coated with goat anti-mouse Igs (Cappel, West Chester, PA) for 15 min at room temperature as previously described (14,31). Recovered cells were washed with PBS and resuspended in RPMI 1640 (Life Technologies) containing 10% normal human serum (NHS).…”

“…T cell differentiation was assessed employing an in vitro, multiple-step differentiation system as previously described (14,32). Briefly, freshly isolated, naive CD4 T cells (CD4 ϩ CD45RA ϩ ) (0.5 ϫ 10 6 /ml) were primed by plate-bound OKT3 (1 g/ml) and anti-CD28 mAb (1 g/ml) in the presence of rIL-2 (10 U/ml), in the presence or the absence of A77 1726, and where indicated in the presence of 50 M UTP (Amersham Pharmacia Biotech, Piscataway, NJ).…”

“…In some experiments freshly isolated naive CD4 T cells were primed in the presence of IL-12 (40 ng/ml) and anti-IL-4 (10 g/ml; Th1-inducing conditions) or in the presence of IL-4 (31.5 ng/ml) and anti-IL-12 (100 ng/ml; Th2-inducing conditions). After 5 days of priming, the cells were harvested, washed, counted, and rested for 2.5 days at 37°C at a concentration of 1 ϫ 10 6 cells/ml in medium supplemented with a nonmitogenic concentration of rIL-2 (10 U/ml) to maintain cell viability (14). Differentiated CD4 T cells were harvested, washed, and assessed for their ability to produce cytokines by intracellular flow cytometry.…”

“…Several in vivo and in vitro experiments emphasize the role of the intensity of T cell stimulation in the outcome of Th differentiation. For example, in an Ag-independent in vitro system, increasing TCR stimulation with a mAb to the ⑀-chain of the TCR complex in the presence of constant costimulation enhanced IFN-␥ production of memory CD4 T effectors and Th1 differentiation in a dose-dependent manner (14). Likewise, high doses of Ag (15), a high affinity of peptide/MHC class II/TCR interactions (16,17), and increasing intensities of Ag presentation (18) favor Th1 differentiation.…”