The phenotype of type 1 and type 2 CD8⁺ T cells activated in vitro is affected by culture conditions and correlates with effector activity

Abstract

Summary We used various culture conditions to generate type 1 (Tc1) or type 2 (Tc2) cytotoxic T cells in vitro. T‐cell receptor (TCR) transgenic T cells were cultured with antigen and spleen cells, or antigen and dendritic cells (DC), or anti‐CD3 and anti‐CD28. Tc1 cultures contained interleukin (IL)‐2 and IL‐6, and Tc2 cultures contained IL‐2, IL‐6 and IL‐4. Tc2 cells generated in each culture condition acquired a CD62Llow CD44high phenotype, had high cytotoxic activity, and secreted IL‐4, IL‐5 and moderate amounts of interferon‐γ (IFN‐γ). In contrast, the phenotype and function of Tc1 cells varied depending on culture conditions. Tc1 cells from anti‐CD3 and anti‐CD28 cultures had high cytotoxic activity and were CD62Llow CD44high, while Tc1 cells from antigen and spleen cell cultures had low cytotoxic activity and were CD62Lhigh CD44low. Tc1 cells from antigen and DC cultures had an intermediate phenotype. All Tc1 cells secreted high amounts of IFN‐γ, but only Tc1 from anti‐CD3 and anti‐CD28 cultures had antitumour activity in vivo. Differences were not caused by suboptimal culture conditions, as Tc1 cells divided at a similar rate whether cultured with antigen and spleen cells or with anti‐CD3 and anti‐CD28. We conclude that IL‐4 not only induces ‘type 2’ cytokine secretion in CD8+ T cells, but also affects their expression of surface markers and cytotoxic activity.

IFN-γ Inhibits IL-4–Induced Type 2 Cytokine Expression by CD8 T Cells In Vivo and Modulates the Anti-Tumor Response

“…1C). This pattern of expression is similar to that reported previously for type 1 and type 2 polarized CD8 T cells generated in vitro by activation with peptide-coated splenocytes in cytokine-polarizing conditions (18). Following overnight restimulation of purified Va2 + cells with anti-receptor Ab, upregulation of expression of mRNAs encoding the type 2 cytokines IL-4, IL-5, IL-10, and IL-13 was evident in both groups exposed to IL-4-expressing tumors; however, expression of mRNA for type 2 cytokines and the type 2 transcription factor GATA-3 was more prominent in the absence of IFN-g ( Fig.…”

Interferon-γ and interleukin-4 reciprocally regulate CD8 expression in CD8+T cells

“…A number of earlier studies have shown that IL-4 induces both the de novo production of IL-4 and down-regulation of CD8 expression in activated CD8 ϩ T cells (12)(13)(14)22). Important clinical correlates have been reported in HIV infection and chronic B cell lymphocytic leukemia, in which IL-4-expressing CD8 ϩ T cells with reduced CD8 expression have been described and may be associated with disease progression (23)(24)(25).…”

Effector CD8+ T cells activated in vitro confer immediate and long‐term tumor protection in vivo

“…In contrast, other studies found that CD62L lo sorted, ex vivo activated effectors, transferred without concomitant vaccination or cytokines, were more effective at inducing tumor rejection than unsorted cells [7,8], and that infiltration of CD62L lo T cells correlated with an objective response against the tumors [7,20,21]. Similarly, we have previously reported that anti-tumor activity correlated with the presence of CD62L lo cells in the adoptively transferred CD8 1 T-cell populations [6]. Several factors may explain these contrasting observations.…”

“…Our culture conditions also used IL-2, but this was only for a short period of time, and only after T cells had already undergone full activation in the presence of Ag. Co-culture with BM-DC, instead of the frequently used spleen APC [5,[7][8][9]19] Line318 T cells activated in vitro in the presence of spleen APC have weak anti-tumor activity in vivo [6], suggesting that the type of APC is important in determining the activity of the resulting T cells. Further experiments are in progress to determine the culture conditions that are critical to the generation of our powerful anti-tumor T cells.…”

“…While former studies by ourselves [6] and other authors [7,8] point to CD62L lo effector memory T (Tem) cells as the only population able to infiltrate and destroy tumors, recent evidence suggests that CD62L hi central memory T (Tcm) cells are superior to Tem cells in suppressing tumor growth when administered in combination with vaccination and exogenous IL-2 [9], and that this better anti-tumor activity correlates with enhanced survival in vivo.…”