Limits...
Restoration of CD28 expression in CD28- CD8+ memory effector T cells reconstitutes antigen-induced IL-2 production.

Topp MS, Riddell SR, Akatsuka Y, Jensen MC, Blattman JN, Greenberg PD - J. Exp. Med. (2003)

Bottom Line: This reflects in part a requirement of activated effector CD8+ T cells for paracrine IL-2 production as a growth and survival factor.Analysis of the fraction of memory CD8+ T cells that continue to express CD28 revealed these cells retain the ability to produce IL-2.These results suggest that the loss of CD28 expression during differentiation of memory/effector CD8+ T cells represents a decisive step in establishing regulation of responding CD8+ T cells, increasing the dependence on CD4+ Th for proliferation after target recognition, and has implications for the treatment of viral disease with adoptively transferred CD8+ T cells.

View Article: PubMed Central - PubMed

Affiliation: Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.

ABSTRACT
The control of many persistent viral infections by Ag-specific cytolytic CD8+ T cells requires a concurrent virus-specific CD4+ Th cell response. This reflects in part a requirement of activated effector CD8+ T cells for paracrine IL-2 production as a growth and survival factor. In human CMV and HIV infection, the majority of differentiated virus-specific CD8+ T cells notably lose the ability to produce IL-2 but also lose expression of CD28, a costimulatory molecule. Analysis of the fraction of memory CD8+ T cells that continue to express CD28 revealed these cells retain the ability to produce IL-2. Therefore, we examined if IL-2 production by CD28- CD8+ T cells could be restored by introduction of a constitutively expressed CD28 gene. Expression of CD28 in CD28- CD8+ CMV- and HIV-specific CD8+ T cells reconstituted the ability to produce IL-2, which could sustain an autocrine proliferative response after Ag recognition. These results suggest that the loss of CD28 expression during differentiation of memory/effector CD8+ T cells represents a decisive step in establishing regulation of responding CD8+ T cells, increasing the dependence on CD4+ Th for proliferation after target recognition, and has implications for the treatment of viral disease with adoptively transferred CD8+ T cells.

Show MeSH

Related in: MedlinePlus

IL-2 production and proliferation of CD28-transduced HIV-specific CD8+ T cell clone after Ag stimulation requires an intact CD28 costimulatory signal. (A) IL-2 production by parental and CD28-transduced HIV-specific CD8+ T cells. HLA A*0301-positive LCLs pulsed with HIVgag/20–28 were incubated with the parental (○ and □) or CD28-transduced (• and ▪) HIV-specific CD8+ T cell clones TL-52-H8 (○ and •) and SM-28A2 (□ and ▪). CTLA-4–Ig was added to indicated wells. The data shown is the mean of two samples for each clone. (B) Parental TL-52-H8 (white bar) and the CD28-transduced clone (black bar) were stimulated with HLA A*0301+ LCLs alone or pulsed with the HIVgag/20–28 peptide. Selected wells were supplemented with CTLA-4–Ig and/or IL-2. Proliferation was assessed by incorporation of [3H]-TdR. The data represents the mean [3H]-TdR incorporation from triplicate wells and is representative of three experiments. (C) The HIV-specific CD28− CD8+ T cell clone SM-28A2 was transduced with either the intact CD28 or a mutant CD28 gene, and CD28 surface expression was assessed. (D) IL-2 production was quantified after Ag stimulation of the parental, and CD28− and CD28 mutant-transduced SM-28A2. HLA A*0301+ APCs were pulsed with the HIVgag/20–28 peptide and incubated with either the parental (white bar), the CD28-transduced (black bar), or the CD28 mutant-transduced (gray bar) HIV-specific CD8+ T cell clone. Supernatants were harvested at 36 h, and IL-2 was measured by ELISA. The data represents the means of two experiments.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2194206&req=5

fig5: IL-2 production and proliferation of CD28-transduced HIV-specific CD8+ T cell clone after Ag stimulation requires an intact CD28 costimulatory signal. (A) IL-2 production by parental and CD28-transduced HIV-specific CD8+ T cells. HLA A*0301-positive LCLs pulsed with HIVgag/20–28 were incubated with the parental (○ and □) or CD28-transduced (• and ▪) HIV-specific CD8+ T cell clones TL-52-H8 (○ and •) and SM-28A2 (□ and ▪). CTLA-4–Ig was added to indicated wells. The data shown is the mean of two samples for each clone. (B) Parental TL-52-H8 (white bar) and the CD28-transduced clone (black bar) were stimulated with HLA A*0301+ LCLs alone or pulsed with the HIVgag/20–28 peptide. Selected wells were supplemented with CTLA-4–Ig and/or IL-2. Proliferation was assessed by incorporation of [3H]-TdR. The data represents the mean [3H]-TdR incorporation from triplicate wells and is representative of three experiments. (C) The HIV-specific CD28− CD8+ T cell clone SM-28A2 was transduced with either the intact CD28 or a mutant CD28 gene, and CD28 surface expression was assessed. (D) IL-2 production was quantified after Ag stimulation of the parental, and CD28− and CD28 mutant-transduced SM-28A2. HLA A*0301+ APCs were pulsed with the HIVgag/20–28 peptide and incubated with either the parental (white bar), the CD28-transduced (black bar), or the CD28 mutant-transduced (gray bar) HIV-specific CD8+ T cell clone. Supernatants were harvested at 36 h, and IL-2 was measured by ELISA. The data represents the means of two experiments.

Mentions: One hallmark of HIV infection is the loss of virus-specific CD4+ Th cells, which normally can provide IL-2 as a paracrine growth factor for responding CD8+ T cells. The majority (70–90%) of HIV-specific CD8+ T cells present during chronic infection have lost CD28 expression (17), and this skewing might contribute to a quantitatively inadequate response and ineffective control of viremia. Therefore, we evaluated if the loss of IL-2 production in HIV-specific CD28− CD8+ T cells represented a multitude of events acquired during infection or if restoring CD28 expression could reestablish the ability of these effector cells to produce sufficient IL-2 to overcome the dependence on CD4+ Th cells. CD28 was expressed in HIV-specific CD28− CD8+ CTL clones by retroviral transduction, and the ability to produce IL-2 was assessed. The parental clones did not produce IL-2 in response to Ag stimulation, whereas the CD28-transduced T cell clones acquired this capacity (Fig. 5 A). The IL-2 production was again abrogated by addition of CTLA-4–Ig (Fig. 5 A).


Restoration of CD28 expression in CD28- CD8+ memory effector T cells reconstitutes antigen-induced IL-2 production.

Topp MS, Riddell SR, Akatsuka Y, Jensen MC, Blattman JN, Greenberg PD - J. Exp. Med. (2003)

IL-2 production and proliferation of CD28-transduced HIV-specific CD8+ T cell clone after Ag stimulation requires an intact CD28 costimulatory signal. (A) IL-2 production by parental and CD28-transduced HIV-specific CD8+ T cells. HLA A*0301-positive LCLs pulsed with HIVgag/20–28 were incubated with the parental (○ and □) or CD28-transduced (• and ▪) HIV-specific CD8+ T cell clones TL-52-H8 (○ and •) and SM-28A2 (□ and ▪). CTLA-4–Ig was added to indicated wells. The data shown is the mean of two samples for each clone. (B) Parental TL-52-H8 (white bar) and the CD28-transduced clone (black bar) were stimulated with HLA A*0301+ LCLs alone or pulsed with the HIVgag/20–28 peptide. Selected wells were supplemented with CTLA-4–Ig and/or IL-2. Proliferation was assessed by incorporation of [3H]-TdR. The data represents the mean [3H]-TdR incorporation from triplicate wells and is representative of three experiments. (C) The HIV-specific CD28− CD8+ T cell clone SM-28A2 was transduced with either the intact CD28 or a mutant CD28 gene, and CD28 surface expression was assessed. (D) IL-2 production was quantified after Ag stimulation of the parental, and CD28− and CD28 mutant-transduced SM-28A2. HLA A*0301+ APCs were pulsed with the HIVgag/20–28 peptide and incubated with either the parental (white bar), the CD28-transduced (black bar), or the CD28 mutant-transduced (gray bar) HIV-specific CD8+ T cell clone. Supernatants were harvested at 36 h, and IL-2 was measured by ELISA. The data represents the means of two experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2194206&req=5

fig5: IL-2 production and proliferation of CD28-transduced HIV-specific CD8+ T cell clone after Ag stimulation requires an intact CD28 costimulatory signal. (A) IL-2 production by parental and CD28-transduced HIV-specific CD8+ T cells. HLA A*0301-positive LCLs pulsed with HIVgag/20–28 were incubated with the parental (○ and □) or CD28-transduced (• and ▪) HIV-specific CD8+ T cell clones TL-52-H8 (○ and •) and SM-28A2 (□ and ▪). CTLA-4–Ig was added to indicated wells. The data shown is the mean of two samples for each clone. (B) Parental TL-52-H8 (white bar) and the CD28-transduced clone (black bar) were stimulated with HLA A*0301+ LCLs alone or pulsed with the HIVgag/20–28 peptide. Selected wells were supplemented with CTLA-4–Ig and/or IL-2. Proliferation was assessed by incorporation of [3H]-TdR. The data represents the mean [3H]-TdR incorporation from triplicate wells and is representative of three experiments. (C) The HIV-specific CD28− CD8+ T cell clone SM-28A2 was transduced with either the intact CD28 or a mutant CD28 gene, and CD28 surface expression was assessed. (D) IL-2 production was quantified after Ag stimulation of the parental, and CD28− and CD28 mutant-transduced SM-28A2. HLA A*0301+ APCs were pulsed with the HIVgag/20–28 peptide and incubated with either the parental (white bar), the CD28-transduced (black bar), or the CD28 mutant-transduced (gray bar) HIV-specific CD8+ T cell clone. Supernatants were harvested at 36 h, and IL-2 was measured by ELISA. The data represents the means of two experiments.
Mentions: One hallmark of HIV infection is the loss of virus-specific CD4+ Th cells, which normally can provide IL-2 as a paracrine growth factor for responding CD8+ T cells. The majority (70–90%) of HIV-specific CD8+ T cells present during chronic infection have lost CD28 expression (17), and this skewing might contribute to a quantitatively inadequate response and ineffective control of viremia. Therefore, we evaluated if the loss of IL-2 production in HIV-specific CD28− CD8+ T cells represented a multitude of events acquired during infection or if restoring CD28 expression could reestablish the ability of these effector cells to produce sufficient IL-2 to overcome the dependence on CD4+ Th cells. CD28 was expressed in HIV-specific CD28− CD8+ CTL clones by retroviral transduction, and the ability to produce IL-2 was assessed. The parental clones did not produce IL-2 in response to Ag stimulation, whereas the CD28-transduced T cell clones acquired this capacity (Fig. 5 A). The IL-2 production was again abrogated by addition of CTLA-4–Ig (Fig. 5 A).

Bottom Line: This reflects in part a requirement of activated effector CD8+ T cells for paracrine IL-2 production as a growth and survival factor.Analysis of the fraction of memory CD8+ T cells that continue to express CD28 revealed these cells retain the ability to produce IL-2.These results suggest that the loss of CD28 expression during differentiation of memory/effector CD8+ T cells represents a decisive step in establishing regulation of responding CD8+ T cells, increasing the dependence on CD4+ Th for proliferation after target recognition, and has implications for the treatment of viral disease with adoptively transferred CD8+ T cells.

View Article: PubMed Central - PubMed

Affiliation: Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.

ABSTRACT
The control of many persistent viral infections by Ag-specific cytolytic CD8+ T cells requires a concurrent virus-specific CD4+ Th cell response. This reflects in part a requirement of activated effector CD8+ T cells for paracrine IL-2 production as a growth and survival factor. In human CMV and HIV infection, the majority of differentiated virus-specific CD8+ T cells notably lose the ability to produce IL-2 but also lose expression of CD28, a costimulatory molecule. Analysis of the fraction of memory CD8+ T cells that continue to express CD28 revealed these cells retain the ability to produce IL-2. Therefore, we examined if IL-2 production by CD28- CD8+ T cells could be restored by introduction of a constitutively expressed CD28 gene. Expression of CD28 in CD28- CD8+ CMV- and HIV-specific CD8+ T cells reconstituted the ability to produce IL-2, which could sustain an autocrine proliferative response after Ag recognition. These results suggest that the loss of CD28 expression during differentiation of memory/effector CD8+ T cells represents a decisive step in establishing regulation of responding CD8+ T cells, increasing the dependence on CD4+ Th for proliferation after target recognition, and has implications for the treatment of viral disease with adoptively transferred CD8+ T cells.

Show MeSH
Related in: MedlinePlus