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Adoptive transfer of immune cells from glaucomatous mice provokes retinal ganglion cell loss in recipients.

Gramlich OW, Ding QJ, Zhu W, Cook A, Anderson MG, Kuehn MH - Acta Neuropathol Commun (2015)

Bottom Line: Signs of pan-retinal inflammation were not detected.Transferred lymphocytes were detected integrated in the spleen and in the retinal ganglion cell layer of recipient animals, albeit at very low frequencies.Furthermore, we observed cell-cell interaction between transferred T-cells and recipient microglia along with focal microglial activation in recipient eyes.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology and Visual Sciences, The University of Iowa, Iowa City, 52242, IA, USA.

ABSTRACT

Introduction: Several studies have indicated that autoimmune and neuroinflammatory processes contribute to the neurodegeneration of retinal ganglion cells in human glaucoma patients and in animal models. To test the involvement of cellular immune processes in the pathophysiology of retinal ganglion cell degeneration in vivo, we carried out adoptive transfer experiments from two independent genetic mouse models of glaucoma into normal recipient mice.

Results: Our findings indicate that transfer results in a progressive loss of retinal ganglion cells and their axons despite normal intraocular pressure in recipient mice. Signs of pan-retinal inflammation were not detected. Similar findings were obtained following transfer of isolated T-lymphocytes, but not after transfer of splenocytes from immune deficient glaucomatous mice. Transferred lymphocytes were detected integrated in the spleen and in the retinal ganglion cell layer of recipient animals, albeit at very low frequencies. Furthermore, we observed cell-cell interaction between transferred T-cells and recipient microglia along with focal microglial activation in recipient eyes.

Conclusion: This study demonstrates that the pathophysiology of glaucomatous degeneration in the tested animal models includes T-cell mediated events that are capable of causing loss of healthy retinal ganglion cells.

No MeSH data available.


Related in: MedlinePlus

T- and B-cell adoptive transfer. a IOP recordings in nee/Rag1− splenocyte recipients (n=, CD3+ and CD19+ cell recipient animals. Data indicate that the transfer does not influence IOP and remains similar to IOP values of naïve mice. The mean IOP over all groups remains at 12.6 ± 2.8 mmHg (p > 0.5). Data are given as mean ± SD. b RGC density in nee/Rag1− splenocyte recipients and CD19+ or CD3+ lymphocyte recipient mice 4 months after transfer. Transfer of nee CD3+ T-lymphocytes induces a significant loss of RGC in animals (N = 5) when compared to recipients of B6 CD3+ cells (N = 8), to B6 CD19+ lymphocyte recipients (N = 8), or to a naïve control group (N = 7, **p < 0.001). Adoptive transfer of the nee CD19+ fractions results in a modest, but not statistically significant, reduction of RGC in recipients (N = 5, p = 0.12) when compared to the corresponding B6 CD19+ lymphocyte recipients. Transfer of splenocytes from immunodeficient nee/Rag1− donors did not result in loss of RGC in recipients (N = 6) when compared to naïve mice and animals having received lymphocytes from B6 donors (p < 0.84). Accordingly, RGC density is also significantly lower in nee when compared to nee/Rag1− animals (##p < 0.001). Triangles represent individual RGC data and the horizontal line designates the group average
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Fig3: T- and B-cell adoptive transfer. a IOP recordings in nee/Rag1− splenocyte recipients (n=, CD3+ and CD19+ cell recipient animals. Data indicate that the transfer does not influence IOP and remains similar to IOP values of naïve mice. The mean IOP over all groups remains at 12.6 ± 2.8 mmHg (p > 0.5). Data are given as mean ± SD. b RGC density in nee/Rag1− splenocyte recipients and CD19+ or CD3+ lymphocyte recipient mice 4 months after transfer. Transfer of nee CD3+ T-lymphocytes induces a significant loss of RGC in animals (N = 5) when compared to recipients of B6 CD3+ cells (N = 8), to B6 CD19+ lymphocyte recipients (N = 8), or to a naïve control group (N = 7, **p < 0.001). Adoptive transfer of the nee CD19+ fractions results in a modest, but not statistically significant, reduction of RGC in recipients (N = 5, p = 0.12) when compared to the corresponding B6 CD19+ lymphocyte recipients. Transfer of splenocytes from immunodeficient nee/Rag1− donors did not result in loss of RGC in recipients (N = 6) when compared to naïve mice and animals having received lymphocytes from B6 donors (p < 0.84). Accordingly, RGC density is also significantly lower in nee when compared to nee/Rag1− animals (##p < 0.001). Triangles represent individual RGC data and the horizontal line designates the group average

Mentions: Following our observation that RGC loss is inducible by adoptive transfer of splenocytes, we sought to determine if the lymphocytes included in these preparations are primarily responsible for the observed RGC loss in recipients and, if so, whether this effect is mediated by B- or T-cells. In order to address this question, we created immunodeficient nee mice by crossing them with Rag1− mice [38]. The resultant immunodeficient nee (nee/Rag1−) lack mature T-cells or B-cells, but display the glaucomatous phenotype with high IOP (25.2 mmHg) and progressive RGC loss. Splenocytes were obtained from these animals and transferred into B6 recipients (N = 6) as described above. In addition, respective CD3 and CD19 positive cell fractions were isolated by flow cytometry from splenocytes of nee mice or B6 donors. Recipient B6 mice were injected with either 1×106 CD3+ T-lymphocytes, or 1.5×106 CD19+ B-lymphocytes from either nee (N = 5, ea.) or healthy B6 donor animals (N = 8, ea.). As noted following splenocyte transfer, adoptive transfer of splenocytes or T- or B-lymphocytes from glaucomatous mice did not increase IOP in the recipient animals. The average IOP of recipient mice was 12.3 ± 2.6 mmHg, similar to the normal population average (13.5 ± 2.5, p < 0.5, Fig. 3a).Fig. 3


Adoptive transfer of immune cells from glaucomatous mice provokes retinal ganglion cell loss in recipients.

Gramlich OW, Ding QJ, Zhu W, Cook A, Anderson MG, Kuehn MH - Acta Neuropathol Commun (2015)

T- and B-cell adoptive transfer. a IOP recordings in nee/Rag1− splenocyte recipients (n=, CD3+ and CD19+ cell recipient animals. Data indicate that the transfer does not influence IOP and remains similar to IOP values of naïve mice. The mean IOP over all groups remains at 12.6 ± 2.8 mmHg (p > 0.5). Data are given as mean ± SD. b RGC density in nee/Rag1− splenocyte recipients and CD19+ or CD3+ lymphocyte recipient mice 4 months after transfer. Transfer of nee CD3+ T-lymphocytes induces a significant loss of RGC in animals (N = 5) when compared to recipients of B6 CD3+ cells (N = 8), to B6 CD19+ lymphocyte recipients (N = 8), or to a naïve control group (N = 7, **p < 0.001). Adoptive transfer of the nee CD19+ fractions results in a modest, but not statistically significant, reduction of RGC in recipients (N = 5, p = 0.12) when compared to the corresponding B6 CD19+ lymphocyte recipients. Transfer of splenocytes from immunodeficient nee/Rag1− donors did not result in loss of RGC in recipients (N = 6) when compared to naïve mice and animals having received lymphocytes from B6 donors (p < 0.84). Accordingly, RGC density is also significantly lower in nee when compared to nee/Rag1− animals (##p < 0.001). Triangles represent individual RGC data and the horizontal line designates the group average
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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Fig3: T- and B-cell adoptive transfer. a IOP recordings in nee/Rag1− splenocyte recipients (n=, CD3+ and CD19+ cell recipient animals. Data indicate that the transfer does not influence IOP and remains similar to IOP values of naïve mice. The mean IOP over all groups remains at 12.6 ± 2.8 mmHg (p > 0.5). Data are given as mean ± SD. b RGC density in nee/Rag1− splenocyte recipients and CD19+ or CD3+ lymphocyte recipient mice 4 months after transfer. Transfer of nee CD3+ T-lymphocytes induces a significant loss of RGC in animals (N = 5) when compared to recipients of B6 CD3+ cells (N = 8), to B6 CD19+ lymphocyte recipients (N = 8), or to a naïve control group (N = 7, **p < 0.001). Adoptive transfer of the nee CD19+ fractions results in a modest, but not statistically significant, reduction of RGC in recipients (N = 5, p = 0.12) when compared to the corresponding B6 CD19+ lymphocyte recipients. Transfer of splenocytes from immunodeficient nee/Rag1− donors did not result in loss of RGC in recipients (N = 6) when compared to naïve mice and animals having received lymphocytes from B6 donors (p < 0.84). Accordingly, RGC density is also significantly lower in nee when compared to nee/Rag1− animals (##p < 0.001). Triangles represent individual RGC data and the horizontal line designates the group average
Mentions: Following our observation that RGC loss is inducible by adoptive transfer of splenocytes, we sought to determine if the lymphocytes included in these preparations are primarily responsible for the observed RGC loss in recipients and, if so, whether this effect is mediated by B- or T-cells. In order to address this question, we created immunodeficient nee mice by crossing them with Rag1− mice [38]. The resultant immunodeficient nee (nee/Rag1−) lack mature T-cells or B-cells, but display the glaucomatous phenotype with high IOP (25.2 mmHg) and progressive RGC loss. Splenocytes were obtained from these animals and transferred into B6 recipients (N = 6) as described above. In addition, respective CD3 and CD19 positive cell fractions were isolated by flow cytometry from splenocytes of nee mice or B6 donors. Recipient B6 mice were injected with either 1×106 CD3+ T-lymphocytes, or 1.5×106 CD19+ B-lymphocytes from either nee (N = 5, ea.) or healthy B6 donor animals (N = 8, ea.). As noted following splenocyte transfer, adoptive transfer of splenocytes or T- or B-lymphocytes from glaucomatous mice did not increase IOP in the recipient animals. The average IOP of recipient mice was 12.3 ± 2.6 mmHg, similar to the normal population average (13.5 ± 2.5, p < 0.5, Fig. 3a).Fig. 3

Bottom Line: Signs of pan-retinal inflammation were not detected.Transferred lymphocytes were detected integrated in the spleen and in the retinal ganglion cell layer of recipient animals, albeit at very low frequencies.Furthermore, we observed cell-cell interaction between transferred T-cells and recipient microglia along with focal microglial activation in recipient eyes.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology and Visual Sciences, The University of Iowa, Iowa City, 52242, IA, USA.

ABSTRACT

Introduction: Several studies have indicated that autoimmune and neuroinflammatory processes contribute to the neurodegeneration of retinal ganglion cells in human glaucoma patients and in animal models. To test the involvement of cellular immune processes in the pathophysiology of retinal ganglion cell degeneration in vivo, we carried out adoptive transfer experiments from two independent genetic mouse models of glaucoma into normal recipient mice.

Results: Our findings indicate that transfer results in a progressive loss of retinal ganglion cells and their axons despite normal intraocular pressure in recipient mice. Signs of pan-retinal inflammation were not detected. Similar findings were obtained following transfer of isolated T-lymphocytes, but not after transfer of splenocytes from immune deficient glaucomatous mice. Transferred lymphocytes were detected integrated in the spleen and in the retinal ganglion cell layer of recipient animals, albeit at very low frequencies. Furthermore, we observed cell-cell interaction between transferred T-cells and recipient microglia along with focal microglial activation in recipient eyes.

Conclusion: This study demonstrates that the pathophysiology of glaucomatous degeneration in the tested animal models includes T-cell mediated events that are capable of causing loss of healthy retinal ganglion cells.

No MeSH data available.


Related in: MedlinePlus