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(Investigative Ophthalmology and Visual Science. 2004;45:857-862.)
© 2004 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.03-1028
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Central Immunotolerance in Transgenic Mice Expressing a Foreign Antigen under Control of the Rhodopsin Promoter

Don-Il Ham,1,2,3 Stephen J. Kim,1,2,4 Jun Chen,1,2 Barbara P. Vistica,1 Robert N. Fariss,5 Robert S. Lee,6 Eric F. Wawrousek,6 Hiroshi Takase,1 Cheng-Rong Yu,1 Charles E. Egwuagu,1 Chi-Chao Chan,1 and Igal Gery1

1From the Laboratories of Immunology, 5Mechanisms of Ocular Diseases, and 6Molecular and Developmental Biology, National Eye Institute, National Institutes of Health, Bethesda, Maryland; the 3Department of Ophthalmology, Sungkyunkwan University School of Medicine, Seoul, Korea; and the 4Howard Hughes Medical Institute-NIH Research Scholars Program, Bethesda, Maryland.

PURPOSE. Different conclusions have been reached in recent studies concerning the immune response or tolerance of transgenic (Tg) mice expressing foreign antigens under control of retinal antigen promoters. The present study was aimed at analyzing the state of tolerance in Tg mice expressing hen egg lysozyme (HEL) under control of the rhodopsin promoter.

METHODS. Tg mice expressing HEL under control of the rhodopsin promoter (RhHEL-Tg) were generated and tested by conventional methods for immune responses against HEL. These Tg mice were also mated with Tg mice expressing HEL-specific receptor on their T lymphocytes and the double-Tg mice were examined for increased apoptosis in their thymi by the TUNEL assay, as well as for loss of HEL-specific T cells, by flow cytometry and proliferative response. The presence of HEL mRNA in mouse thymi was determined by RT-PCR.

RESULTS. RhHEL-Tg mice developed tolerance to HEL, shown by reduced cellular and humoral responses to HEL, as well as by the failure of ocular inflammation to develop after immunization with HEL. RhHEL-Tg mice expressed HEL mRNA in their thymus, and the tolerogenic mechanism in these mice was shown to be thymic deletion of HEL-specific T cells by the following observations in the double-Tg mice: (1) increased apoptosis in their thymi, (2) remarkable reduction in the proportion of the HEL-specific T cells, and (3) loss of lymphocyte response to low concentrations of HEL.

CONCLUSIONS. Tg mice expressing HEL under control of the rhodopsin promoter develop a tolerance for the foreign antigen, apparently as a result of thymic expression of HEL and deletion of T cells specific to this antigen.




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