Limits...
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

Appearance of ocular microglia in recipients of B6 or nee lymphocytes. a Iba1 immunostained microglia in the whole mounted retina of an age-matched naïve control mouse depicting quiescent, ramified microglia. Retinal microglia in recipients of (b) CD19+ and (c) CD3+ cells donated from B6 mice appear uniformly ramified 28 days after transfer. d Horizontal sections of the optic nerve also did not reveal signs of microglial activation. Here the optic nerve of a recipient after adoptive transfer of B6 CD19+ cell fraction is shown. Numerous activated microglia are readily identified in recipients of (e) CD19+ or (f) CD3+ lymphocytes from nee donor mice (bottom row). Both early-stage microglial activation, indicated by hypertrophic somatic areas (arrows in f), and highly activated microglia (arrowhead in e) were observed. g Interactions between transferred T-lymphocytes and activated resident microglia are occasionally found. These cells are located outside the retinal vasculature (V) and are accompanied by additional, unidentified, DAPI positive endogenous cells. h Microglial activation was not observed in the optic nerve of any recipients. Scale bars in a-c, e-f is 50 μm and in d, h 20 μm
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4591529&req=5

Fig5: Appearance of ocular microglia in recipients of B6 or nee lymphocytes. a Iba1 immunostained microglia in the whole mounted retina of an age-matched naïve control mouse depicting quiescent, ramified microglia. Retinal microglia in recipients of (b) CD19+ and (c) CD3+ cells donated from B6 mice appear uniformly ramified 28 days after transfer. d Horizontal sections of the optic nerve also did not reveal signs of microglial activation. Here the optic nerve of a recipient after adoptive transfer of B6 CD19+ cell fraction is shown. Numerous activated microglia are readily identified in recipients of (e) CD19+ or (f) CD3+ lymphocytes from nee donor mice (bottom row). Both early-stage microglial activation, indicated by hypertrophic somatic areas (arrows in f), and highly activated microglia (arrowhead in e) were observed. g Interactions between transferred T-lymphocytes and activated resident microglia are occasionally found. These cells are located outside the retinal vasculature (V) and are accompanied by additional, unidentified, DAPI positive endogenous cells. h Microglial activation was not observed in the optic nerve of any recipients. Scale bars in a-c, e-f is 50 μm and in d, h 20 μm

Mentions: One of the hallmarks of neurodegeneration, including retinal degeneration, is the activation of microglia [35, 41]. As the innate immune cell of the central nervous system, microglia strongly interface with the systemic immune system to cause bidirectional signaling, into and out of the CNS [42]. Hence, we examined if retinal microglia are affected by splenocyte transfer and whether there is evidence of their interaction with transferred cells in the retinas of recipient mice using the microglia marker Iba1 [29]. In the healthy retina microglial cells have a highly ramified phenotype with small soma, thin branches and a fine network of cellular processes. An early stage of microglial activation is indicated by retraction of their branches and hypertrophic somas, whereas fully activated microglia cells are characterized by an amoeboid shape [31]. It was apparent that only retinae of nee lymphocyte recipients, but not B6 lymphocyte recipients, contain a small population of hypertrophic Iba1+ microglia with an amoeboid shape as well as a few highly activated microglia (Fig. 5). Those microglia that appeared to be activated are frequently observed in close contact to transferred DsRed/CD3+ cells from nee mice even if quiescent microglia are apparent a short distance away. This interaction between transferred lymphocytes and resident microglia is occasionally accompanied by other, not yet identified, endogenous cells (Fig. 5f). These observations suggest an interaction between CD3+ T-lymphocytes and resident microglia.Fig. 5


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)

Appearance of ocular microglia in recipients of B6 or nee lymphocytes. a Iba1 immunostained microglia in the whole mounted retina of an age-matched naïve control mouse depicting quiescent, ramified microglia. Retinal microglia in recipients of (b) CD19+ and (c) CD3+ cells donated from B6 mice appear uniformly ramified 28 days after transfer. d Horizontal sections of the optic nerve also did not reveal signs of microglial activation. Here the optic nerve of a recipient after adoptive transfer of B6 CD19+ cell fraction is shown. Numerous activated microglia are readily identified in recipients of (e) CD19+ or (f) CD3+ lymphocytes from nee donor mice (bottom row). Both early-stage microglial activation, indicated by hypertrophic somatic areas (arrows in f), and highly activated microglia (arrowhead in e) were observed. g Interactions between transferred T-lymphocytes and activated resident microglia are occasionally found. These cells are located outside the retinal vasculature (V) and are accompanied by additional, unidentified, DAPI positive endogenous cells. h Microglial activation was not observed in the optic nerve of any recipients. Scale bars in a-c, e-f is 50 μm and in d, h 20 μm
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4591529&req=5

Fig5: Appearance of ocular microglia in recipients of B6 or nee lymphocytes. a Iba1 immunostained microglia in the whole mounted retina of an age-matched naïve control mouse depicting quiescent, ramified microglia. Retinal microglia in recipients of (b) CD19+ and (c) CD3+ cells donated from B6 mice appear uniformly ramified 28 days after transfer. d Horizontal sections of the optic nerve also did not reveal signs of microglial activation. Here the optic nerve of a recipient after adoptive transfer of B6 CD19+ cell fraction is shown. Numerous activated microglia are readily identified in recipients of (e) CD19+ or (f) CD3+ lymphocytes from nee donor mice (bottom row). Both early-stage microglial activation, indicated by hypertrophic somatic areas (arrows in f), and highly activated microglia (arrowhead in e) were observed. g Interactions between transferred T-lymphocytes and activated resident microglia are occasionally found. These cells are located outside the retinal vasculature (V) and are accompanied by additional, unidentified, DAPI positive endogenous cells. h Microglial activation was not observed in the optic nerve of any recipients. Scale bars in a-c, e-f is 50 μm and in d, h 20 μm
Mentions: One of the hallmarks of neurodegeneration, including retinal degeneration, is the activation of microglia [35, 41]. As the innate immune cell of the central nervous system, microglia strongly interface with the systemic immune system to cause bidirectional signaling, into and out of the CNS [42]. Hence, we examined if retinal microglia are affected by splenocyte transfer and whether there is evidence of their interaction with transferred cells in the retinas of recipient mice using the microglia marker Iba1 [29]. In the healthy retina microglial cells have a highly ramified phenotype with small soma, thin branches and a fine network of cellular processes. An early stage of microglial activation is indicated by retraction of their branches and hypertrophic somas, whereas fully activated microglia cells are characterized by an amoeboid shape [31]. It was apparent that only retinae of nee lymphocyte recipients, but not B6 lymphocyte recipients, contain a small population of hypertrophic Iba1+ microglia with an amoeboid shape as well as a few highly activated microglia (Fig. 5). Those microglia that appeared to be activated are frequently observed in close contact to transferred DsRed/CD3+ cells from nee mice even if quiescent microglia are apparent a short distance away. This interaction between transferred lymphocytes and resident microglia is occasionally accompanied by other, not yet identified, endogenous cells (Fig. 5f). These observations suggest an interaction between CD3+ T-lymphocytes and resident microglia.Fig. 5

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