Susceptibilty of murine corneal kpithklial and endothelial cells to lysis by complement-dependent alloantibody

S. L. Hargrave, E. S. Mayhem, S. Hedge, J. Y. Niederkorn

Research output: Contribution to journalArticle

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Abstract

Purpose. To determine if orthotopic corneal allografts stimulated the production of allospecific cuotoxic antibody and if comeal epithelial and endothelial cells are vulnerable to K sis by complement-dependent antibody. Methods. The capacity of orthotopic corneal alIngrafts to induce allospecific antibody was determined by transplanting C3H/Hej (H-2k) corneal grafts onto CB6FI (H-2h'd) mice. Grafted mice were bled 30 days later and the serum tested by ELISA for the presence of anti-C3H/Hej alloantibody. Anti-C3H alloantiserum was prepared by immunizing CB6F1 (H-2 ) mice with two intramuscular injections of 5x10 allogeneic C3H/Hej {H-2 ) spleen cells. Complement-dependent antibody-mediated lysis of C3H corneal epithelial cells and C3H corneal endothelial cells was determined in a conventional in vitro 4-hr " Cr-release assay in the presence and absence of 50% human aqueous humor (AH). Results. Orthotopic corneal allografts induced significant allospecific IgG antibody production in 5 of 7 animals tested (P<0.05). in three separate experiments, hyper-immune anti-C3H/Hej serum produced 21% to 50% cytolysis of C3H/Hej corneal endothetial cells. By contrast, the same antiserum failed to l\se OH/Hej corneal epithelial cells. Antibody-mediated lysis of comeal epithelial cells could be inhibited b> the addition of 50% human AH. Conclusions. Although orthotopic corneal allografts can elicit the production of alloantibodies. comeal epithelial cells are resistant to antibody-mediated lysis. The resistance to lysis by complement-dependent antibody is presumably due to the expression of complement decay accelerating factor (DAF) on the corneal epithelium. By contrast, murine corneal endothetial cells do not express DAF and as a result are susceptible to complement-mediated lysis. However, DAF present in the AH protects corneal endothelial cells from injury inflicted by complement dependent cytolytic antibody.

Original languageEnglish (US)
JournalInvestigative Ophthalmology and Visual Science
Volume38
Issue number4
StatePublished - 1997

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Isoantibodies
Endothelial Cells
CD55 Antigens
Antibodies
Aqueous Humor
Allografts
Epithelial Cells
Corneal Epithelium
Intramuscular Injections
Antibody Formation
Spleen
Immunoglobulin G
Enzyme-Linked Immunosorbent Assay
Transplants
Wounds and Injuries
Serum

ASJC Scopus subject areas

  • Ophthalmology

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Susceptibilty of murine corneal kpithklial and endothelial cells to lysis by complement-dependent alloantibody. / Hargrave, S. L.; Mayhem, E. S.; Hedge, S.; Niederkorn, J. Y.

In: Investigative Ophthalmology and Visual Science, Vol. 38, No. 4, 1997.

Research output: Contribution to journalArticle

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title = "Susceptibilty of murine corneal kpithklial and endothelial cells to lysis by complement-dependent alloantibody",
abstract = "Purpose. To determine if orthotopic corneal allografts stimulated the production of allospecific cuotoxic antibody and if comeal epithelial and endothelial cells are vulnerable to K sis by complement-dependent antibody. Methods. The capacity of orthotopic corneal alIngrafts to induce allospecific antibody was determined by transplanting C3H/Hej (H-2k) corneal grafts onto CB6FI (H-2h'd) mice. Grafted mice were bled 30 days later and the serum tested by ELISA for the presence of anti-C3H/Hej alloantibody. Anti-C3H alloantiserum was prepared by immunizing CB6F1 (H-2 ) mice with two intramuscular injections of 5x10 allogeneic C3H/Hej {H-2 ) spleen cells. Complement-dependent antibody-mediated lysis of C3H corneal epithelial cells and C3H corneal endothelial cells was determined in a conventional in vitro 4-hr {"} Cr-release assay in the presence and absence of 50{\%} human aqueous humor (AH). Results. Orthotopic corneal allografts induced significant allospecific IgG antibody production in 5 of 7 animals tested (P<0.05). in three separate experiments, hyper-immune anti-C3H/Hej serum produced 21{\%} to 50{\%} cytolysis of C3H/Hej corneal endothetial cells. By contrast, the same antiserum failed to l\se OH/Hej corneal epithelial cells. Antibody-mediated lysis of comeal epithelial cells could be inhibited b> the addition of 50{\%} human AH. Conclusions. Although orthotopic corneal allografts can elicit the production of alloantibodies. comeal epithelial cells are resistant to antibody-mediated lysis. The resistance to lysis by complement-dependent antibody is presumably due to the expression of complement decay accelerating factor (DAF) on the corneal epithelium. By contrast, murine corneal endothetial cells do not express DAF and as a result are susceptible to complement-mediated lysis. However, DAF present in the AH protects corneal endothelial cells from injury inflicted by complement dependent cytolytic antibody.",
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AU - Hargrave, S. L.

AU - Mayhem, E. S.

AU - Hedge, S.

AU - Niederkorn, J. Y.

PY - 1997

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N2 - Purpose. To determine if orthotopic corneal allografts stimulated the production of allospecific cuotoxic antibody and if comeal epithelial and endothelial cells are vulnerable to K sis by complement-dependent antibody. Methods. The capacity of orthotopic corneal alIngrafts to induce allospecific antibody was determined by transplanting C3H/Hej (H-2k) corneal grafts onto CB6FI (H-2h'd) mice. Grafted mice were bled 30 days later and the serum tested by ELISA for the presence of anti-C3H/Hej alloantibody. Anti-C3H alloantiserum was prepared by immunizing CB6F1 (H-2 ) mice with two intramuscular injections of 5x10 allogeneic C3H/Hej {H-2 ) spleen cells. Complement-dependent antibody-mediated lysis of C3H corneal epithelial cells and C3H corneal endothelial cells was determined in a conventional in vitro 4-hr " Cr-release assay in the presence and absence of 50% human aqueous humor (AH). Results. Orthotopic corneal allografts induced significant allospecific IgG antibody production in 5 of 7 animals tested (P<0.05). in three separate experiments, hyper-immune anti-C3H/Hej serum produced 21% to 50% cytolysis of C3H/Hej corneal endothetial cells. By contrast, the same antiserum failed to l\se OH/Hej corneal epithelial cells. Antibody-mediated lysis of comeal epithelial cells could be inhibited b> the addition of 50% human AH. Conclusions. Although orthotopic corneal allografts can elicit the production of alloantibodies. comeal epithelial cells are resistant to antibody-mediated lysis. The resistance to lysis by complement-dependent antibody is presumably due to the expression of complement decay accelerating factor (DAF) on the corneal epithelium. By contrast, murine corneal endothetial cells do not express DAF and as a result are susceptible to complement-mediated lysis. However, DAF present in the AH protects corneal endothelial cells from injury inflicted by complement dependent cytolytic antibody.

AB - Purpose. To determine if orthotopic corneal allografts stimulated the production of allospecific cuotoxic antibody and if comeal epithelial and endothelial cells are vulnerable to K sis by complement-dependent antibody. Methods. The capacity of orthotopic corneal alIngrafts to induce allospecific antibody was determined by transplanting C3H/Hej (H-2k) corneal grafts onto CB6FI (H-2h'd) mice. Grafted mice were bled 30 days later and the serum tested by ELISA for the presence of anti-C3H/Hej alloantibody. Anti-C3H alloantiserum was prepared by immunizing CB6F1 (H-2 ) mice with two intramuscular injections of 5x10 allogeneic C3H/Hej {H-2 ) spleen cells. Complement-dependent antibody-mediated lysis of C3H corneal epithelial cells and C3H corneal endothelial cells was determined in a conventional in vitro 4-hr " Cr-release assay in the presence and absence of 50% human aqueous humor (AH). Results. Orthotopic corneal allografts induced significant allospecific IgG antibody production in 5 of 7 animals tested (P<0.05). in three separate experiments, hyper-immune anti-C3H/Hej serum produced 21% to 50% cytolysis of C3H/Hej corneal endothetial cells. By contrast, the same antiserum failed to l\se OH/Hej corneal epithelial cells. Antibody-mediated lysis of comeal epithelial cells could be inhibited b> the addition of 50% human AH. Conclusions. Although orthotopic corneal allografts can elicit the production of alloantibodies. comeal epithelial cells are resistant to antibody-mediated lysis. The resistance to lysis by complement-dependent antibody is presumably due to the expression of complement decay accelerating factor (DAF) on the corneal epithelium. By contrast, murine corneal endothetial cells do not express DAF and as a result are susceptible to complement-mediated lysis. However, DAF present in the AH protects corneal endothelial cells from injury inflicted by complement dependent cytolytic antibody.

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