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Br J Ophthalmol 1999;83:973-979 doi:10.1136/bjo.83.8.973
  • Original Article
    • Laboratory science

Suppression of induction of experimental immune mediated blepharoconjunctivitis by tolerogenic conjugates of the antigen and monomethoxypolyethylene glycol

  1. Atsuki Fukushimaa,
  2. Koji Nishinoa,
  3. Hironori Yoshidaa,
  4. Masaru Takatab,
  5. Hisayuki Uenoa
  1. aDepartment of Ophthalmology, Kochi Medical School, Nankoku City, Japan, bDepartment of Immunology, Kochi Medical School, Nankoku City, Japan
  1. Atsuki Fukushima, Laboratory of Immunology, Department of Ophthalmology, Kochi Medical School, Kohasu, Oko-cho, Nankoku 783–8505, Japan.
  • Accepted 15 March 1999

Abstract

AIM Covalent conjugates consisting of diverse antigens coupled to optimal numbers of monomethoxypolyethylene glycol (mPEG) molecules have been shown to suppress antigen specific antibody formation. In this study, the possibility was examined that the same conjugates might prevent experimental immune mediated blepharoconjunctivitis (EC, formerly EAC) which had been shown to be caused by CD4+ T cells—that is, to cell mediated immunity.

METHODS 6–8 week old male Lewis rats were used. The test groups of rats received two intravenous injections, each of 300 μg, of a conjugate of ovalbumin mPEG (OVA(mPEG)11) in phosphate buffered saline (PBS), 14 and 28 days before the single immunisation with OVA in complete Freund’s adjuvant. The rats were challenged 3 weeks later by eye drops containing OVA; 24 hours later they were sacrificed, and their eyes, blood, and lymph nodes were harvested for histological examination and determination of anti-OVA antibody titres and levels of cellular immunity. Two control groups received PBS or OVA in PBS before immunisation. Furthermore, the possibility that OVA(mPEG)11may have induced OVA specific suppressor cells was tested by establishing the effects of the co-transfer of splenocytes from OVA(mPEG)11 treated rats with OVA primed lymph node cells on the manifestations of EC.

RESULTS Either PBS or OVA pretreated rats, which had not received OVA(mPEG)11, developed high levels of antibodies and cell mediated immune responses to OVA, and application of eye drops led to blepharoconjunctivitis with massive cellular infiltration. In contrast, pretreatment with OVA(mPEG)11 prevented cellular infiltration into the lids and conjunctivas, as well as the formation of detectable humoral and cellular immunity against OVA. Co-transfer of splenocytes from OVA(mPEG)11 treated rats with OVA primed lymph node cells suppressed the cellular infiltration on application of OVA on the conjunctiva.

CONCLUSIONS These data indicate that intravenous injection of OVA(mPEG)11conjugates suppressed both humoral and cellular immunity by the effects of antigen specific suppressor cells, thus leading to the inhibition of development of EC.

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