Expression of major histocompatibility complex molecules in rodent retina

Immunohistochemical study

Jie Zhang, Guey Shuang Wu, Sei Ichi Ishimoto, Geeta Pararajasegaram, Narsing A. Rao

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

Purpose. In Lewis rats, S-antigen-induced intraocular inflammation, which occurs initially in the retina, is mediated by T cells requiring major histocompatibility complex (MHC)-restricted antigen presentation. In such organ-specific inflammation, antigen presentation may take place at the site of the initial inflammatory response. In the present study an attempt waS made to determine the presence of the putative antigen-presenting cells in the retina of rats. Methods. Six bone marrow chimeras were constructed by transferring 50 x 106 donor [Lewis x Brown Norway, (LBN) FI] bone marrow cells into lethally irradiated Brown Norway rats. Three chimeras, 3 Lewis, and 2 Brown Norway rats each received intravenous injections of gamma interferon (IFN-γ) at a dose of 2 X 105 U/rat, 48 and 24 hours before enucleation of the globes. Enucleated globes from the 3 remaining untreated chimeras, 21 additional Lewis rats, and 6 Brown Norway rats served as controls. Retinas from all globes were prepared for either wholemount or cryosectioning and were stained using various primary antibodies, including anti-Lewis MHC class II (OX3), anti-rat MHC class II (OX6), anti-Lewis MHC class I (II-69), anti-rat MHC class I (OX18), anti-macrophage complement receptor 3 (OX42), anti-monocytes/macrophages (ED1, ED2, and EB3), and anti- glial fibrillary acidic protein (GFAP). Fluorescein-conjugated goat anti- mouse and rhodamine-conjugated anti-rabbit immunoglobulins were used to detect the monoclonal or polyclonal antibodies, respectively. All the specimens were examined under Zeiss confocal laser scanning microscopy. The positively stained cells were counted for quantitative analysis. Results. Major histocompatibility complex class II (OX6)-positive cells were demonstrated in the wholemount retinas of IFN-γ-untreated chimeras, Lewis, and Brown Norway rats. These cells showed a dendritic morphology and increased significantly in number in IFN-γ-treated Lewis and Brown Norway rats. Expression of Lewis-specific class I (II-69) and class II (OX3) molecules was detected in a few perivascular cells in the retina of chimeric rats treated with IFN-γ. Most dendritic cells in the retina expressed the macrophage markers, ED1 and OX42, without IFN-γ treatment. However, vascular endothelia, retinal pigment epithelia, Muller cells, and astrocytes stained neither by Class II molecules nor by macrophage markers. The vascular endothelium and retinal pigment epithelium was found to express constitutively class I molecules (OX18). Conclusions. A subpopulation of retinal microglia can express MHC class II molecules. Only a few of these are derived from bone marrow. Retinal microglial cells, particularly those derived from bone marrow, may participate in antigen presentation and the subsequent development of retinitis, as seen in S-antigen-induced experimental autoimmune uveoretinitis.

Original languageEnglish (US)
Pages (from-to)1848-1857
Number of pages10
JournalInvestigative Ophthalmology and Visual Science
Volume38
Issue number9
StatePublished - Aug 1997
Externally publishedYes

Fingerprint

Major Histocompatibility Complex
Retina
Rodentia
Interferons
Antigen Presentation
Macrophages
Retinal Pigment Epithelium
Bone Marrow
Vascular Endothelium
Cryoultramicrotomy
Macrophage-1 Antigen
Inflammation
Ependymoglial Cells
Retinitis
Antigens
Rhodamines
Glial Fibrillary Acidic Protein
Microglia
Antigen-Presenting Cells
Norway

All Science Journal Classification (ASJC) codes

  • Ophthalmology

Cite this

Expression of major histocompatibility complex molecules in rodent retina : Immunohistochemical study. / Zhang, Jie; Wu, Guey Shuang; Ishimoto, Sei Ichi; Pararajasegaram, Geeta; Rao, Narsing A.

In: Investigative Ophthalmology and Visual Science, Vol. 38, No. 9, 08.1997, p. 1848-1857.

Research output: Contribution to journalArticle

Zhang, Jie ; Wu, Guey Shuang ; Ishimoto, Sei Ichi ; Pararajasegaram, Geeta ; Rao, Narsing A. / Expression of major histocompatibility complex molecules in rodent retina : Immunohistochemical study. In: Investigative Ophthalmology and Visual Science. 1997 ; Vol. 38, No. 9. pp. 1848-1857.
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title = "Expression of major histocompatibility complex molecules in rodent retina: Immunohistochemical study",
abstract = "Purpose. In Lewis rats, S-antigen-induced intraocular inflammation, which occurs initially in the retina, is mediated by T cells requiring major histocompatibility complex (MHC)-restricted antigen presentation. In such organ-specific inflammation, antigen presentation may take place at the site of the initial inflammatory response. In the present study an attempt waS made to determine the presence of the putative antigen-presenting cells in the retina of rats. Methods. Six bone marrow chimeras were constructed by transferring 50 x 106 donor [Lewis x Brown Norway, (LBN) FI] bone marrow cells into lethally irradiated Brown Norway rats. Three chimeras, 3 Lewis, and 2 Brown Norway rats each received intravenous injections of gamma interferon (IFN-γ) at a dose of 2 X 105 U/rat, 48 and 24 hours before enucleation of the globes. Enucleated globes from the 3 remaining untreated chimeras, 21 additional Lewis rats, and 6 Brown Norway rats served as controls. Retinas from all globes were prepared for either wholemount or cryosectioning and were stained using various primary antibodies, including anti-Lewis MHC class II (OX3), anti-rat MHC class II (OX6), anti-Lewis MHC class I (II-69), anti-rat MHC class I (OX18), anti-macrophage complement receptor 3 (OX42), anti-monocytes/macrophages (ED1, ED2, and EB3), and anti- glial fibrillary acidic protein (GFAP). Fluorescein-conjugated goat anti- mouse and rhodamine-conjugated anti-rabbit immunoglobulins were used to detect the monoclonal or polyclonal antibodies, respectively. All the specimens were examined under Zeiss confocal laser scanning microscopy. The positively stained cells were counted for quantitative analysis. Results. Major histocompatibility complex class II (OX6)-positive cells were demonstrated in the wholemount retinas of IFN-γ-untreated chimeras, Lewis, and Brown Norway rats. These cells showed a dendritic morphology and increased significantly in number in IFN-γ-treated Lewis and Brown Norway rats. Expression of Lewis-specific class I (II-69) and class II (OX3) molecules was detected in a few perivascular cells in the retina of chimeric rats treated with IFN-γ. Most dendritic cells in the retina expressed the macrophage markers, ED1 and OX42, without IFN-γ treatment. However, vascular endothelia, retinal pigment epithelia, Muller cells, and astrocytes stained neither by Class II molecules nor by macrophage markers. The vascular endothelium and retinal pigment epithelium was found to express constitutively class I molecules (OX18). Conclusions. A subpopulation of retinal microglia can express MHC class II molecules. Only a few of these are derived from bone marrow. Retinal microglial cells, particularly those derived from bone marrow, may participate in antigen presentation and the subsequent development of retinitis, as seen in S-antigen-induced experimental autoimmune uveoretinitis.",
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T1 - Expression of major histocompatibility complex molecules in rodent retina

T2 - Immunohistochemical study

AU - Zhang, Jie

AU - Wu, Guey Shuang

AU - Ishimoto, Sei Ichi

AU - Pararajasegaram, Geeta

AU - Rao, Narsing A.

PY - 1997/8

Y1 - 1997/8

N2 - Purpose. In Lewis rats, S-antigen-induced intraocular inflammation, which occurs initially in the retina, is mediated by T cells requiring major histocompatibility complex (MHC)-restricted antigen presentation. In such organ-specific inflammation, antigen presentation may take place at the site of the initial inflammatory response. In the present study an attempt waS made to determine the presence of the putative antigen-presenting cells in the retina of rats. Methods. Six bone marrow chimeras were constructed by transferring 50 x 106 donor [Lewis x Brown Norway, (LBN) FI] bone marrow cells into lethally irradiated Brown Norway rats. Three chimeras, 3 Lewis, and 2 Brown Norway rats each received intravenous injections of gamma interferon (IFN-γ) at a dose of 2 X 105 U/rat, 48 and 24 hours before enucleation of the globes. Enucleated globes from the 3 remaining untreated chimeras, 21 additional Lewis rats, and 6 Brown Norway rats served as controls. Retinas from all globes were prepared for either wholemount or cryosectioning and were stained using various primary antibodies, including anti-Lewis MHC class II (OX3), anti-rat MHC class II (OX6), anti-Lewis MHC class I (II-69), anti-rat MHC class I (OX18), anti-macrophage complement receptor 3 (OX42), anti-monocytes/macrophages (ED1, ED2, and EB3), and anti- glial fibrillary acidic protein (GFAP). Fluorescein-conjugated goat anti- mouse and rhodamine-conjugated anti-rabbit immunoglobulins were used to detect the monoclonal or polyclonal antibodies, respectively. All the specimens were examined under Zeiss confocal laser scanning microscopy. The positively stained cells were counted for quantitative analysis. Results. Major histocompatibility complex class II (OX6)-positive cells were demonstrated in the wholemount retinas of IFN-γ-untreated chimeras, Lewis, and Brown Norway rats. These cells showed a dendritic morphology and increased significantly in number in IFN-γ-treated Lewis and Brown Norway rats. Expression of Lewis-specific class I (II-69) and class II (OX3) molecules was detected in a few perivascular cells in the retina of chimeric rats treated with IFN-γ. Most dendritic cells in the retina expressed the macrophage markers, ED1 and OX42, without IFN-γ treatment. However, vascular endothelia, retinal pigment epithelia, Muller cells, and astrocytes stained neither by Class II molecules nor by macrophage markers. The vascular endothelium and retinal pigment epithelium was found to express constitutively class I molecules (OX18). Conclusions. A subpopulation of retinal microglia can express MHC class II molecules. Only a few of these are derived from bone marrow. Retinal microglial cells, particularly those derived from bone marrow, may participate in antigen presentation and the subsequent development of retinitis, as seen in S-antigen-induced experimental autoimmune uveoretinitis.

AB - Purpose. In Lewis rats, S-antigen-induced intraocular inflammation, which occurs initially in the retina, is mediated by T cells requiring major histocompatibility complex (MHC)-restricted antigen presentation. In such organ-specific inflammation, antigen presentation may take place at the site of the initial inflammatory response. In the present study an attempt waS made to determine the presence of the putative antigen-presenting cells in the retina of rats. Methods. Six bone marrow chimeras were constructed by transferring 50 x 106 donor [Lewis x Brown Norway, (LBN) FI] bone marrow cells into lethally irradiated Brown Norway rats. Three chimeras, 3 Lewis, and 2 Brown Norway rats each received intravenous injections of gamma interferon (IFN-γ) at a dose of 2 X 105 U/rat, 48 and 24 hours before enucleation of the globes. Enucleated globes from the 3 remaining untreated chimeras, 21 additional Lewis rats, and 6 Brown Norway rats served as controls. Retinas from all globes were prepared for either wholemount or cryosectioning and were stained using various primary antibodies, including anti-Lewis MHC class II (OX3), anti-rat MHC class II (OX6), anti-Lewis MHC class I (II-69), anti-rat MHC class I (OX18), anti-macrophage complement receptor 3 (OX42), anti-monocytes/macrophages (ED1, ED2, and EB3), and anti- glial fibrillary acidic protein (GFAP). Fluorescein-conjugated goat anti- mouse and rhodamine-conjugated anti-rabbit immunoglobulins were used to detect the monoclonal or polyclonal antibodies, respectively. All the specimens were examined under Zeiss confocal laser scanning microscopy. The positively stained cells were counted for quantitative analysis. Results. Major histocompatibility complex class II (OX6)-positive cells were demonstrated in the wholemount retinas of IFN-γ-untreated chimeras, Lewis, and Brown Norway rats. These cells showed a dendritic morphology and increased significantly in number in IFN-γ-treated Lewis and Brown Norway rats. Expression of Lewis-specific class I (II-69) and class II (OX3) molecules was detected in a few perivascular cells in the retina of chimeric rats treated with IFN-γ. Most dendritic cells in the retina expressed the macrophage markers, ED1 and OX42, without IFN-γ treatment. However, vascular endothelia, retinal pigment epithelia, Muller cells, and astrocytes stained neither by Class II molecules nor by macrophage markers. The vascular endothelium and retinal pigment epithelium was found to express constitutively class I molecules (OX18). Conclusions. A subpopulation of retinal microglia can express MHC class II molecules. Only a few of these are derived from bone marrow. Retinal microglial cells, particularly those derived from bone marrow, may participate in antigen presentation and the subsequent development of retinitis, as seen in S-antigen-induced experimental autoimmune uveoretinitis.

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