The immunohistochemical detection of bone marrow derived Ia+ cells in the chimeric rodent retina

Jie Zhang, G. S. Wu, S. Ishimoto, N. A. Rao

Research output: Contribution to journalArticle

Abstract

Purpose. Previous adoptive T-cell transfer studies using chimeric rats suggested presentation of S-antigen by bone marrow derived cells. The present study attempts to reveal the Ia- antigen expressing bone marrow-derived cells, their localization and their phenotype in the bone marrow chimeric rat retina. Methods. The chimeras were constructed by transferring 50x106 donor (Lewis x Brown Norway F1) bone marrow cells into lethally irradiated Brown Norway (BN) rats. Three chimeras and 3 Lewis rats received intravenous injection of IFN-γ (2x105 U/rat/day, x2). Three chimeras, 3 Lewis rats and 3 BN rats served as controls. The retinas from all the animals were prepared for whole mount immunofluorescent stain, utilizing various primary antibodies (1:100): anti-Ia (OX6); anti-la Lewis specific (OX3); anti-MHC class I antigens (OX18), anti-Lewis MHC class I (IL-69); anti-tissue macrophages (ED1, ED2, and ED3); anti-macrophage/microglia FC receptor (OX42) and anti-GFAP. The positively stained cells were counted for quantitative analysis. Results. All IFN-γ treated rats revealed the presence of OX6+, OX3+, OX42+, ED1+ and ED3+ cells in the retina. OX6+ cells were present in the control chimeric rat retinas but could not be detected in the other two controls. In contrast OX3+ cells were seen in the retina of IFN-γ treated animals only. These cells were distributed perivascularily and had morphologic similarities of ED1+ cells. The OX3+ cells were smaller than OX 42+ cells. Moreover, the OX3+ cells were significantly fewer in number than OX6- or OX42+ cells. OX18- and IL-69+ staining was seen in the retinal vasculature of all rats. Conclusions. A distinct subpopulation of Ia+ cells with dendritic morphology (OX3+), derived from bone marrow, are found in the retinal perivascular sites. These perivascular cells may present retinal antigens to induce organ-specific autoimmune uveoretinitis.

Original languageEnglish (US)
JournalInvestigative Ophthalmology and Visual Science
Volume37
Issue number3
StatePublished - Feb 15 1996
Externally publishedYes

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Retina
Rodentia
Bone Marrow
Bone Marrow Cells
Macrophages
Histocompatibility Antigens Class I
Adoptive Transfer
Histocompatibility Antigens Class II
Antigen Presentation
Microglia
Norway
Intravenous Injections
Dendritic Cells
Coloring Agents
Staining and Labeling
T-Lymphocytes
Phenotype
Antigens
Antibodies

All Science Journal Classification (ASJC) codes

  • Ophthalmology

Cite this

The immunohistochemical detection of bone marrow derived Ia+ cells in the chimeric rodent retina. / Zhang, Jie; Wu, G. S.; Ishimoto, S.; Rao, N. A.

In: Investigative Ophthalmology and Visual Science, Vol. 37, No. 3, 15.02.1996.

Research output: Contribution to journalArticle

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abstract = "Purpose. Previous adoptive T-cell transfer studies using chimeric rats suggested presentation of S-antigen by bone marrow derived cells. The present study attempts to reveal the Ia- antigen expressing bone marrow-derived cells, their localization and their phenotype in the bone marrow chimeric rat retina. Methods. The chimeras were constructed by transferring 50x106 donor (Lewis x Brown Norway F1) bone marrow cells into lethally irradiated Brown Norway (BN) rats. Three chimeras and 3 Lewis rats received intravenous injection of IFN-γ (2x105 U/rat/day, x2). Three chimeras, 3 Lewis rats and 3 BN rats served as controls. The retinas from all the animals were prepared for whole mount immunofluorescent stain, utilizing various primary antibodies (1:100): anti-Ia (OX6); anti-la Lewis specific (OX3); anti-MHC class I antigens (OX18), anti-Lewis MHC class I (IL-69); anti-tissue macrophages (ED1, ED2, and ED3); anti-macrophage/microglia FC receptor (OX42) and anti-GFAP. The positively stained cells were counted for quantitative analysis. Results. All IFN-γ treated rats revealed the presence of OX6+, OX3+, OX42+, ED1+ and ED3+ cells in the retina. OX6+ cells were present in the control chimeric rat retinas but could not be detected in the other two controls. In contrast OX3+ cells were seen in the retina of IFN-γ treated animals only. These cells were distributed perivascularily and had morphologic similarities of ED1+ cells. The OX3+ cells were smaller than OX 42+ cells. Moreover, the OX3+ cells were significantly fewer in number than OX6- or OX42+ cells. OX18- and IL-69+ staining was seen in the retinal vasculature of all rats. Conclusions. A distinct subpopulation of Ia+ cells with dendritic morphology (OX3+), derived from bone marrow, are found in the retinal perivascular sites. These perivascular cells may present retinal antigens to induce organ-specific autoimmune uveoretinitis.",
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N2 - Purpose. Previous adoptive T-cell transfer studies using chimeric rats suggested presentation of S-antigen by bone marrow derived cells. The present study attempts to reveal the Ia- antigen expressing bone marrow-derived cells, their localization and their phenotype in the bone marrow chimeric rat retina. Methods. The chimeras were constructed by transferring 50x106 donor (Lewis x Brown Norway F1) bone marrow cells into lethally irradiated Brown Norway (BN) rats. Three chimeras and 3 Lewis rats received intravenous injection of IFN-γ (2x105 U/rat/day, x2). Three chimeras, 3 Lewis rats and 3 BN rats served as controls. The retinas from all the animals were prepared for whole mount immunofluorescent stain, utilizing various primary antibodies (1:100): anti-Ia (OX6); anti-la Lewis specific (OX3); anti-MHC class I antigens (OX18), anti-Lewis MHC class I (IL-69); anti-tissue macrophages (ED1, ED2, and ED3); anti-macrophage/microglia FC receptor (OX42) and anti-GFAP. The positively stained cells were counted for quantitative analysis. Results. All IFN-γ treated rats revealed the presence of OX6+, OX3+, OX42+, ED1+ and ED3+ cells in the retina. OX6+ cells were present in the control chimeric rat retinas but could not be detected in the other two controls. In contrast OX3+ cells were seen in the retina of IFN-γ treated animals only. These cells were distributed perivascularily and had morphologic similarities of ED1+ cells. The OX3+ cells were smaller than OX 42+ cells. Moreover, the OX3+ cells were significantly fewer in number than OX6- or OX42+ cells. OX18- and IL-69+ staining was seen in the retinal vasculature of all rats. Conclusions. A distinct subpopulation of Ia+ cells with dendritic morphology (OX3+), derived from bone marrow, are found in the retinal perivascular sites. These perivascular cells may present retinal antigens to induce organ-specific autoimmune uveoretinitis.

AB - Purpose. Previous adoptive T-cell transfer studies using chimeric rats suggested presentation of S-antigen by bone marrow derived cells. The present study attempts to reveal the Ia- antigen expressing bone marrow-derived cells, their localization and their phenotype in the bone marrow chimeric rat retina. Methods. The chimeras were constructed by transferring 50x106 donor (Lewis x Brown Norway F1) bone marrow cells into lethally irradiated Brown Norway (BN) rats. Three chimeras and 3 Lewis rats received intravenous injection of IFN-γ (2x105 U/rat/day, x2). Three chimeras, 3 Lewis rats and 3 BN rats served as controls. The retinas from all the animals were prepared for whole mount immunofluorescent stain, utilizing various primary antibodies (1:100): anti-Ia (OX6); anti-la Lewis specific (OX3); anti-MHC class I antigens (OX18), anti-Lewis MHC class I (IL-69); anti-tissue macrophages (ED1, ED2, and ED3); anti-macrophage/microglia FC receptor (OX42) and anti-GFAP. The positively stained cells were counted for quantitative analysis. Results. All IFN-γ treated rats revealed the presence of OX6+, OX3+, OX42+, ED1+ and ED3+ cells in the retina. OX6+ cells were present in the control chimeric rat retinas but could not be detected in the other two controls. In contrast OX3+ cells were seen in the retina of IFN-γ treated animals only. These cells were distributed perivascularily and had morphologic similarities of ED1+ cells. The OX3+ cells were smaller than OX 42+ cells. Moreover, the OX3+ cells were significantly fewer in number than OX6- or OX42+ cells. OX18- and IL-69+ staining was seen in the retinal vasculature of all rats. Conclusions. A distinct subpopulation of Ia+ cells with dendritic morphology (OX3+), derived from bone marrow, are found in the retinal perivascular sites. These perivascular cells may present retinal antigens to induce organ-specific autoimmune uveoretinitis.

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