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Peripheral expression of Jak3 is required to maintain T lymphocyte function.

Thomis DC, Berg LJ - J. Exp. Med. (1997)

Bottom Line: The Jak family tyrosine kinase, Jak3, is involved in signaling through cytokine receptors that utilize the common gamma chain (gammac), such as those for IL-2, IL-4, IL-7, IL-9, and IL-15.Jak3 expression in the thymus restores normal T cell development, including CD8+, gammadelta, and natural killer cells.However, the loss of Jak3 protein in peripheral T cells leads to the Jak3-/- phenotype, demonstrating that Jak3 is constitutively required to maintain T cell function.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts 02138, USA.

ABSTRACT
The Jak family tyrosine kinase, Jak3, is involved in signaling through cytokine receptors that utilize the common gamma chain (gammac), such as those for IL-2, IL-4, IL-7, IL-9, and IL-15. Recent studies of Jak3-deficient mice and humans have demonstrated that Jak3 plays a critical role in B and T lymphocyte maturation and function. The T lymphocyte defects in Jak3-deficient mice include a small thymus, a decrease in peripheral CD8+ cells, an increase in the surface expression of activation markers, and a severe reduction in proliferative and cytokine secretion responses to mitogenic stimuli. To determine whether the peripheral T lymphocyte defects result from aberrant maturation in the thymus or from the absence of Jak3 protein in peripheral T cells, we generated reconstituted mice that express normal levels of Jak3 protein in the thymus but lose Jak3 expression in peripheral T cells. Jak3 expression in the thymus restores normal T cell development, including CD8+, gammadelta, and natural killer cells. However, the loss of Jak3 protein in peripheral T cells leads to the Jak3-/- phenotype, demonstrating that Jak3 is constitutively required to maintain T cell function.

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Functional responses of Jak3−/− (tgthy) T cells are restored in the thymus, but lost in the periphery. (A) Thymocytes and splenocytes from  Jak3+/−, Jak3−/−, Jak3−/− (tgthy+spl), and Jak3−/− (tgthy) mice were analyzed for proliferation (top) and IL-2 (middle) or IL-3 (bottom) secretion in response to stimulation with anti-CD3 plus anti-CD28 antibodies. All data shown are from one 35-d-old Jak3−/− (tgthy+spl) mouse and one 35-d-old Jak3−/−  (tgthy) mouse (designated B). IL-2 secretion data from splenocytes of two additional Jak3−/− (tgthy) mice, one 25 d of age (designated A) and one 42 d of  age (designated C), are also shown. All values are mean ± SD. Overall, no statistically significant differences between Jak3+/−, Jak3−/− (tgthy+spl), and  Jak3−/− (tgthy) thymocytes were observed for IL-2 or IL-3 secretion responses. (B) Thymocytes and splenocytes from Jak3+/−, Jak3−/−, and Jak3−/− (tgkd)  mice were stimulated with antibodies to CD3 plus CD28. Proliferative, IL-2 secretion, and IL-3 secretion responses are shown. All values are mean ± SD.  For proliferation assays, all cell populations cultured in media alone gave responses of <500 cpm. For cytokine assays, cells cultured in media alone secreted undetectable levels of IL-2 (<0.02 U/ml) and IL-3 (<0.01 U/ml). Data are representative of 2–9 independent experiments.
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Figure 4: Functional responses of Jak3−/− (tgthy) T cells are restored in the thymus, but lost in the periphery. (A) Thymocytes and splenocytes from Jak3+/−, Jak3−/−, Jak3−/− (tgthy+spl), and Jak3−/− (tgthy) mice were analyzed for proliferation (top) and IL-2 (middle) or IL-3 (bottom) secretion in response to stimulation with anti-CD3 plus anti-CD28 antibodies. All data shown are from one 35-d-old Jak3−/− (tgthy+spl) mouse and one 35-d-old Jak3−/− (tgthy) mouse (designated B). IL-2 secretion data from splenocytes of two additional Jak3−/− (tgthy) mice, one 25 d of age (designated A) and one 42 d of age (designated C), are also shown. All values are mean ± SD. Overall, no statistically significant differences between Jak3+/−, Jak3−/− (tgthy+spl), and Jak3−/− (tgthy) thymocytes were observed for IL-2 or IL-3 secretion responses. (B) Thymocytes and splenocytes from Jak3+/−, Jak3−/−, and Jak3−/− (tgkd) mice were stimulated with antibodies to CD3 plus CD28. Proliferative, IL-2 secretion, and IL-3 secretion responses are shown. All values are mean ± SD. For proliferation assays, all cell populations cultured in media alone gave responses of <500 cpm. For cytokine assays, cells cultured in media alone secreted undetectable levels of IL-2 (<0.02 U/ml) and IL-3 (<0.01 U/ml). Data are representative of 2–9 independent experiments.

Mentions: T cells were stimulated by culturing in wells coated with goat anti–hamster antibody followed by antiCD3 antibody, in the presence of anti-CD28 antibody hybridoma supernatant as described above. As a control, cells were cultured in media alone. For proliferation assays, 1 × 105 total thymocytes or total splenocytes adjusted to contain 1 × 104 CD4+ T cells per well were cultured for 48 h, then pulsed overnight with [3H]thymidine and counted. For cytokine assays, 1 × 106 total thymocytes or total splenocytes adjusted to contain 1 × 105 CD4+ T cells per well were stimulated. Supernatants from duplicate cultures were harvested at 24 h, and IL-2 and IL-3 levels were quantitated by titration on indicator cells. For IL-2, 1 U/ml corresponds to 1/50 maximal proliferation of the HT-2 indicator cells (Fig. 4 A) or 1/10 maximal proliferation of the indicator cells (Fig. 4 B). For IL-3, 1 U/ml corresponds to 1/10 maximal proliferation of the DA-1 indicator cells.


Peripheral expression of Jak3 is required to maintain T lymphocyte function.

Thomis DC, Berg LJ - J. Exp. Med. (1997)

Functional responses of Jak3−/− (tgthy) T cells are restored in the thymus, but lost in the periphery. (A) Thymocytes and splenocytes from  Jak3+/−, Jak3−/−, Jak3−/− (tgthy+spl), and Jak3−/− (tgthy) mice were analyzed for proliferation (top) and IL-2 (middle) or IL-3 (bottom) secretion in response to stimulation with anti-CD3 plus anti-CD28 antibodies. All data shown are from one 35-d-old Jak3−/− (tgthy+spl) mouse and one 35-d-old Jak3−/−  (tgthy) mouse (designated B). IL-2 secretion data from splenocytes of two additional Jak3−/− (tgthy) mice, one 25 d of age (designated A) and one 42 d of  age (designated C), are also shown. All values are mean ± SD. Overall, no statistically significant differences between Jak3+/−, Jak3−/− (tgthy+spl), and  Jak3−/− (tgthy) thymocytes were observed for IL-2 or IL-3 secretion responses. (B) Thymocytes and splenocytes from Jak3+/−, Jak3−/−, and Jak3−/− (tgkd)  mice were stimulated with antibodies to CD3 plus CD28. Proliferative, IL-2 secretion, and IL-3 secretion responses are shown. All values are mean ± SD.  For proliferation assays, all cell populations cultured in media alone gave responses of <500 cpm. For cytokine assays, cells cultured in media alone secreted undetectable levels of IL-2 (<0.02 U/ml) and IL-3 (<0.01 U/ml). Data are representative of 2–9 independent experiments.
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Related In: Results  -  Collection

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Figure 4: Functional responses of Jak3−/− (tgthy) T cells are restored in the thymus, but lost in the periphery. (A) Thymocytes and splenocytes from Jak3+/−, Jak3−/−, Jak3−/− (tgthy+spl), and Jak3−/− (tgthy) mice were analyzed for proliferation (top) and IL-2 (middle) or IL-3 (bottom) secretion in response to stimulation with anti-CD3 plus anti-CD28 antibodies. All data shown are from one 35-d-old Jak3−/− (tgthy+spl) mouse and one 35-d-old Jak3−/− (tgthy) mouse (designated B). IL-2 secretion data from splenocytes of two additional Jak3−/− (tgthy) mice, one 25 d of age (designated A) and one 42 d of age (designated C), are also shown. All values are mean ± SD. Overall, no statistically significant differences between Jak3+/−, Jak3−/− (tgthy+spl), and Jak3−/− (tgthy) thymocytes were observed for IL-2 or IL-3 secretion responses. (B) Thymocytes and splenocytes from Jak3+/−, Jak3−/−, and Jak3−/− (tgkd) mice were stimulated with antibodies to CD3 plus CD28. Proliferative, IL-2 secretion, and IL-3 secretion responses are shown. All values are mean ± SD. For proliferation assays, all cell populations cultured in media alone gave responses of <500 cpm. For cytokine assays, cells cultured in media alone secreted undetectable levels of IL-2 (<0.02 U/ml) and IL-3 (<0.01 U/ml). Data are representative of 2–9 independent experiments.
Mentions: T cells were stimulated by culturing in wells coated with goat anti–hamster antibody followed by antiCD3 antibody, in the presence of anti-CD28 antibody hybridoma supernatant as described above. As a control, cells were cultured in media alone. For proliferation assays, 1 × 105 total thymocytes or total splenocytes adjusted to contain 1 × 104 CD4+ T cells per well were cultured for 48 h, then pulsed overnight with [3H]thymidine and counted. For cytokine assays, 1 × 106 total thymocytes or total splenocytes adjusted to contain 1 × 105 CD4+ T cells per well were stimulated. Supernatants from duplicate cultures were harvested at 24 h, and IL-2 and IL-3 levels were quantitated by titration on indicator cells. For IL-2, 1 U/ml corresponds to 1/50 maximal proliferation of the HT-2 indicator cells (Fig. 4 A) or 1/10 maximal proliferation of the indicator cells (Fig. 4 B). For IL-3, 1 U/ml corresponds to 1/10 maximal proliferation of the DA-1 indicator cells.

Bottom Line: The Jak family tyrosine kinase, Jak3, is involved in signaling through cytokine receptors that utilize the common gamma chain (gammac), such as those for IL-2, IL-4, IL-7, IL-9, and IL-15.Jak3 expression in the thymus restores normal T cell development, including CD8+, gammadelta, and natural killer cells.However, the loss of Jak3 protein in peripheral T cells leads to the Jak3-/- phenotype, demonstrating that Jak3 is constitutively required to maintain T cell function.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts 02138, USA.

ABSTRACT
The Jak family tyrosine kinase, Jak3, is involved in signaling through cytokine receptors that utilize the common gamma chain (gammac), such as those for IL-2, IL-4, IL-7, IL-9, and IL-15. Recent studies of Jak3-deficient mice and humans have demonstrated that Jak3 plays a critical role in B and T lymphocyte maturation and function. The T lymphocyte defects in Jak3-deficient mice include a small thymus, a decrease in peripheral CD8+ cells, an increase in the surface expression of activation markers, and a severe reduction in proliferative and cytokine secretion responses to mitogenic stimuli. To determine whether the peripheral T lymphocyte defects result from aberrant maturation in the thymus or from the absence of Jak3 protein in peripheral T cells, we generated reconstituted mice that express normal levels of Jak3 protein in the thymus but lose Jak3 expression in peripheral T cells. Jak3 expression in the thymus restores normal T cell development, including CD8+, gammadelta, and natural killer cells. However, the loss of Jak3 protein in peripheral T cells leads to the Jak3-/- phenotype, demonstrating that Jak3 is constitutively required to maintain T cell function.

Show MeSH
Related in: MedlinePlus