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International Immunology Advance Access originally published online on February 10, 2009
International Immunology 2009 21(4):349-360; doi:10.1093/intimm/dxp004
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© The Japanese Society for Immunology. 2009. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org

Immunization of rabbits with DNase II leads to formation of polyclonal antibodies with DNase and RNase activities

Michael A. Krasnorutskii, Valentina N. Buneva and Georgy A. Nevinsky

Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, pr. Lavrent'eva 8, Novosibirsk 630090, Russia

Correspondence to: G. A. Nevinsky; E-mail: nevinsky{at}niboch.nsc.ru

The serum of patients with many autoimmune (AI) diseases contains small fractions of antibodies possessing both DNase and RNase activities. It was shown that immunization of rabbits with DNA, RNA, DNase I and RNase leads to production of antibodies with DNase and RNase activities. It is not known whether anti-idiotypic antibodies against DNase II can possess DNase or RNase activity. Electrophoretically and immunologically homogeneous polyclonal IgGs (pIgGs) from the sera of rabbits immunized with DNase II were obtained by sequential chromatography of the serum proteins on Protein A-Sepharose and gel filtration. It was shown for the first time that immunization of healthy rabbits with bovine DNase II produces IgGs with intrinsic DNase and RNase activities. IgGs from rabbits immunized with BSA or non-immunized animals were catalytically inactive. It was shown that ~10% of the total IgG DNase and RNase activities belong to anti-idiotypic antibodies to DNase II (~0.1% of total pIgGs), while 90% of the activities did not interact with Sepharose bearing antibodies against DNase II and might be antibodies to nucleic acids bound to DNase II. Affinity chromatography on DNA-cellulose using elution of antibodies with different concentration of NaCl and an acidic buffer separated catalytic IgGs into five antibody subfractions, three of which hydrolyzed RNA faster than DNA while two subfractions demonstrated only DNase activity. Our data suggest that a fraction of abzymes from AI patients hydrolyzing both DNA and RNA can contain subfraction of antibodies against DNase II.

Keywords: abzymes, immunization with DNase II, RNA and DNA hydrolysis

Transmitting editor: A. Cooke

Received 1 September 2008, accepted 6 January 2009.


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