OBJECTIVE: To examine the normal human corneal stroma for the presence of bone marrow-derived cells. METHODS: Thirty-four corneas from donors aged 56 to 71 years were used. The stroma of the donor corneas was examined immunohistochemically by fluorescent microscopy. CD45-positive and -negative cells were separated from collagenase-digested stroma by magnetic beads, and the expression of toll-like receptor 4 was analyzed. RESULTS: CD45-positive cells were mainly found in the anterior stroma of the central and paracentral cornea as well as all stromal layers of the peripheral cornea (n = 5). These cells uniformly expressed CD11b, CD11c, CD14, and HLA-DR antigen but not CD3, CD19, CD56, or CD66, indicative of bone marrow-derived monocyte lineage cells, which can include monocytes, macrophages, or dendritic cells. CD45-positive cells isolated with magnetic beads accounted for 6.0% of total stromal cells (n = 20). Stromal CD45-positive cells, but not CD45-negative cells, expressed toll-like receptor 4 by flow cytometry and reverse-transcriptase polymerase chain reaction. CONCLUSION: Our findings demonstrate that DR antigen-positive bone marrow-derived monocyte lineage cells exist in the anterior and peripheral posterior stroma of normal human cornea. Clinical Relevance These cells may play a role in the innate and adaptive immune responses in the human cornea.
OBJECTIVE: To examine the normal humancorneal stroma for the presence of bone marrow-derived cells. METHODS: Thirty-four corneas from donors aged 56 to 71 years were used. The stroma of the donor corneas was examined immunohistochemically by fluorescent microscopy. CD45-positive and -negative cells were separated from collagenase-digested stroma by magnetic beads, and the expression of toll-like receptor 4 was analyzed. RESULTS:CD45-positive cells were mainly found in the anterior stroma of the central and paracentral cornea as well as all stromal layers of the peripheral cornea (n = 5). These cells uniformly expressed CD11b, CD11c, CD14, and HLA-DR antigen but not CD3, CD19, CD56, or CD66, indicative of bone marrow-derived monocyte lineage cells, which can include monocytes, macrophages, or dendritic cells. CD45-positive cells isolated with magnetic beads accounted for 6.0% of total stromal cells (n = 20). Stromal CD45-positive cells, but not CD45-negative cells, expressed toll-like receptor 4 by flow cytometry and reverse-transcriptase polymerase chain reaction. CONCLUSION: Our findings demonstrate that DR antigen-positive bone marrow-derived monocyte lineage cells exist in the anterior and peripheral posterior stroma of normal human cornea. Clinical Relevance These cells may play a role in the innate and adaptive immune responses in the human cornea.
Authors: Tina B McKay; Yashar Seyed-Razavi; Chiara E Ghezzi; Gabriela Dieckmann; Thomas J F Nieland; Dana M Cairns; Rachel E Pollard; Pedram Hamrah; David L Kaplan Journal: Prog Retin Eye Res Date: 2018-11-16 Impact factor: 21.198