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Deletion of IFT20 in early stage T lymphocyte differentiation inhibits the development of collagen-induced arthritis.

Yuan X, Garrett-Sinha LA, Sarkar D, Yang S - Bone Res (2014)

Bottom Line: Recently, IFT20 was found to be also expressed in non-ciliated T cells and have functions in immune synapse formation and signaling in vitro.However, how IFT20 regulates T-cell development and activation in vivo is still unknown.These results demonstrate that deletion of IFT20 in the early stage of T-cell development inhibited CIA development through regulating T-cell development and the expression of critical cytokines.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Biology, School of Dental Medicine, University at Buffalo, The State University of New York , Buffalo, NY 14214, USA.

ABSTRACT
IFT20 is the smallest member of the intraflagellar transport protein (IFT) complex B. It is involved in cilia formation. Studies of IFT20 have been confined to ciliated cells. Recently, IFT20 was found to be also expressed in non-ciliated T cells and have functions in immune synapse formation and signaling in vitro. However, how IFT20 regulates T-cell development and activation in vivo is still unknown. We deleted the IFT20 gene in early and later stages of T-cell development by crossing IFT20(flox/flox) (IFT20(f/f) ) mice with Lck-Cre and CD4-Cre transgenic mice, and investigated the role of IFT20 in T-cell maturation and in the development of T cell-mediated collagen-induced arthritis (CIA). We found that both Lck-Cre/IFT20(f/f) and CD4-Cre/IFT20(f/f) mice were indistinguishable from their wild-type littermates in body size, as well as in the morphology and weight of the spleen and thymus. However, the number of CD4- and CD8-positive cells was significantly lower in thymus and spleen in Lck-Cre/IFT20(f/f) mice. Meanwhile, the incidence and severity of CIA symptoms were significantly decreased, and inflammation in the paw was significantly inhibited in Lck-Cre/IFT20(f/f) mice compared to Lck-Cre/IFT20(+/+) littermates. Deletion IFT20 in more mature T cells of CD4-Cre/IFT20(f/f) mice had only mild effects on the development of T cells and CIA. The expression of IL-1β, IL-6 and TGF-β1 were significantly downregulated in the paw of Lck-Cre/IFT20(f/f) mice, but just slight decreased in CD4-Cre/IFT20(f/f) mice. These results demonstrate that deletion of IFT20 in the early stage of T-cell development inhibited CIA development through regulating T-cell development and the expression of critical cytokines.

No MeSH data available.


Related in: MedlinePlus

Both Lck-Cre/IFT20f/f and CD4-Cre/IFT20f/f mice were indistinguishable from wild-type littermates in mouse body size, and in the morphology and weights of the spleen and thymus. (a) Representative photograph of Lck-Cre/IFT20+/+ and Lck-Cre/IFT20f/f mice at 3 months of age showing similar body mass and appearance. (b) Thymus and spleen from Lck-Cre/IFT20f/f mice shown next to those from a Lck-Cre/IFT20+/+ littermate. Similar Lck-Cre/ IFT20f/f thymic weight was found compared to age-matched Lck-Cre/IFT20+/+ (n=10, P>0.05) (c) Representative photograph of CD4-Cre/IFT20+/+ and CD4-Cre/IFT20f/f mice at 3 months of age showing similar body mass and appearance. (d) Thymus and spleen from CD4-Cre/IFT20f/f mice shown next to those from a CD4-Cre/IFT20+/+ littermate. Similar Lck-Cre/IFT20f/f thymic weight was observed compared to CD4-Cre/IFT20+/+ littermate (n=10, P>0.05) ND, no significant difference.
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Figure 2: Both Lck-Cre/IFT20f/f and CD4-Cre/IFT20f/f mice were indistinguishable from wild-type littermates in mouse body size, and in the morphology and weights of the spleen and thymus. (a) Representative photograph of Lck-Cre/IFT20+/+ and Lck-Cre/IFT20f/f mice at 3 months of age showing similar body mass and appearance. (b) Thymus and spleen from Lck-Cre/IFT20f/f mice shown next to those from a Lck-Cre/IFT20+/+ littermate. Similar Lck-Cre/ IFT20f/f thymic weight was found compared to age-matched Lck-Cre/IFT20+/+ (n=10, P>0.05) (c) Representative photograph of CD4-Cre/IFT20+/+ and CD4-Cre/IFT20f/f mice at 3 months of age showing similar body mass and appearance. (d) Thymus and spleen from CD4-Cre/IFT20f/f mice shown next to those from a CD4-Cre/IFT20+/+ littermate. Similar Lck-Cre/IFT20f/f thymic weight was observed compared to CD4-Cre/IFT20+/+ littermate (n=10, P>0.05) ND, no significant difference.

Mentions: Lck-Cre/IFT20f/f and CD4-cre/IFT20f/f mice were born at normal Mendelian ratios and appeared normal (Figure 2a and 2c). Further analysis of spleen and thymus weights did not reveal any abnormalities. The spleens and thymuses in Lck-Cre/IFT20f/f and CD4-Cre/IFT20f/f mice have similar weights compared to wild-type control littermates (Figure 2b and 2d). Deletion of IFT20 in an early stage of T-cell differentiation reduced the population of CD4- and CD8-positive cells To investigate the role of IFT20 in T-cell development, we detected the CD4- and CD8-positive cells in both thymus and spleen with flow cytometry. As shown in Figure 3a and 3b, depletion of IFT20 in the beginning of DP stage with Lck-Cre resulted in a significant loss of CD4+, CD8+ and CD4+CD8+ T cells in the spleen, while it increased the CD4−CD8− population. In the thymus, the CD4+ and CD8+ T cells were also significantly reduced, accompanied by increased CD4−CD8− cell numbers. However, the numbers of CD4 and CD8 single- and double-positive T cells were not apparently affected when IFT20 was deleted with CD4-Cre (Figure 3c and 3d), indicating that the loss of IFT20 in DP stage does not significantly alter thymocyte differentiation. These findings suggest that IFT20 may function in DN proliferation, but is not required for further differentiation.


Deletion of IFT20 in early stage T lymphocyte differentiation inhibits the development of collagen-induced arthritis.

Yuan X, Garrett-Sinha LA, Sarkar D, Yang S - Bone Res (2014)

Both Lck-Cre/IFT20f/f and CD4-Cre/IFT20f/f mice were indistinguishable from wild-type littermates in mouse body size, and in the morphology and weights of the spleen and thymus. (a) Representative photograph of Lck-Cre/IFT20+/+ and Lck-Cre/IFT20f/f mice at 3 months of age showing similar body mass and appearance. (b) Thymus and spleen from Lck-Cre/IFT20f/f mice shown next to those from a Lck-Cre/IFT20+/+ littermate. Similar Lck-Cre/ IFT20f/f thymic weight was found compared to age-matched Lck-Cre/IFT20+/+ (n=10, P>0.05) (c) Representative photograph of CD4-Cre/IFT20+/+ and CD4-Cre/IFT20f/f mice at 3 months of age showing similar body mass and appearance. (d) Thymus and spleen from CD4-Cre/IFT20f/f mice shown next to those from a CD4-Cre/IFT20+/+ littermate. Similar Lck-Cre/IFT20f/f thymic weight was observed compared to CD4-Cre/IFT20+/+ littermate (n=10, P>0.05) ND, no significant difference.
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Figure 2: Both Lck-Cre/IFT20f/f and CD4-Cre/IFT20f/f mice were indistinguishable from wild-type littermates in mouse body size, and in the morphology and weights of the spleen and thymus. (a) Representative photograph of Lck-Cre/IFT20+/+ and Lck-Cre/IFT20f/f mice at 3 months of age showing similar body mass and appearance. (b) Thymus and spleen from Lck-Cre/IFT20f/f mice shown next to those from a Lck-Cre/IFT20+/+ littermate. Similar Lck-Cre/ IFT20f/f thymic weight was found compared to age-matched Lck-Cre/IFT20+/+ (n=10, P>0.05) (c) Representative photograph of CD4-Cre/IFT20+/+ and CD4-Cre/IFT20f/f mice at 3 months of age showing similar body mass and appearance. (d) Thymus and spleen from CD4-Cre/IFT20f/f mice shown next to those from a CD4-Cre/IFT20+/+ littermate. Similar Lck-Cre/IFT20f/f thymic weight was observed compared to CD4-Cre/IFT20+/+ littermate (n=10, P>0.05) ND, no significant difference.
Mentions: Lck-Cre/IFT20f/f and CD4-cre/IFT20f/f mice were born at normal Mendelian ratios and appeared normal (Figure 2a and 2c). Further analysis of spleen and thymus weights did not reveal any abnormalities. The spleens and thymuses in Lck-Cre/IFT20f/f and CD4-Cre/IFT20f/f mice have similar weights compared to wild-type control littermates (Figure 2b and 2d). Deletion of IFT20 in an early stage of T-cell differentiation reduced the population of CD4- and CD8-positive cells To investigate the role of IFT20 in T-cell development, we detected the CD4- and CD8-positive cells in both thymus and spleen with flow cytometry. As shown in Figure 3a and 3b, depletion of IFT20 in the beginning of DP stage with Lck-Cre resulted in a significant loss of CD4+, CD8+ and CD4+CD8+ T cells in the spleen, while it increased the CD4−CD8− population. In the thymus, the CD4+ and CD8+ T cells were also significantly reduced, accompanied by increased CD4−CD8− cell numbers. However, the numbers of CD4 and CD8 single- and double-positive T cells were not apparently affected when IFT20 was deleted with CD4-Cre (Figure 3c and 3d), indicating that the loss of IFT20 in DP stage does not significantly alter thymocyte differentiation. These findings suggest that IFT20 may function in DN proliferation, but is not required for further differentiation.

Bottom Line: Recently, IFT20 was found to be also expressed in non-ciliated T cells and have functions in immune synapse formation and signaling in vitro.However, how IFT20 regulates T-cell development and activation in vivo is still unknown.These results demonstrate that deletion of IFT20 in the early stage of T-cell development inhibited CIA development through regulating T-cell development and the expression of critical cytokines.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Biology, School of Dental Medicine, University at Buffalo, The State University of New York , Buffalo, NY 14214, USA.

ABSTRACT
IFT20 is the smallest member of the intraflagellar transport protein (IFT) complex B. It is involved in cilia formation. Studies of IFT20 have been confined to ciliated cells. Recently, IFT20 was found to be also expressed in non-ciliated T cells and have functions in immune synapse formation and signaling in vitro. However, how IFT20 regulates T-cell development and activation in vivo is still unknown. We deleted the IFT20 gene in early and later stages of T-cell development by crossing IFT20(flox/flox) (IFT20(f/f) ) mice with Lck-Cre and CD4-Cre transgenic mice, and investigated the role of IFT20 in T-cell maturation and in the development of T cell-mediated collagen-induced arthritis (CIA). We found that both Lck-Cre/IFT20(f/f) and CD4-Cre/IFT20(f/f) mice were indistinguishable from their wild-type littermates in body size, as well as in the morphology and weight of the spleen and thymus. However, the number of CD4- and CD8-positive cells was significantly lower in thymus and spleen in Lck-Cre/IFT20(f/f) mice. Meanwhile, the incidence and severity of CIA symptoms were significantly decreased, and inflammation in the paw was significantly inhibited in Lck-Cre/IFT20(f/f) mice compared to Lck-Cre/IFT20(+/+) littermates. Deletion IFT20 in more mature T cells of CD4-Cre/IFT20(f/f) mice had only mild effects on the development of T cells and CIA. The expression of IL-1β, IL-6 and TGF-β1 were significantly downregulated in the paw of Lck-Cre/IFT20(f/f) mice, but just slight decreased in CD4-Cre/IFT20(f/f) mice. These results demonstrate that deletion of IFT20 in the early stage of T-cell development inhibited CIA development through regulating T-cell development and the expression of critical cytokines.

No MeSH data available.


Related in: MedlinePlus