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Immune Complex Decomplementation of Canine Sera for Use in a Complement-Fixation Test for Diagnosis of Visceral Leishmaniasis^*

Barbara J. Flemmings, Michael G. Pappas, Charlotte Miller Keenan AND Wayne T. Hockmeyer

Canine sera frequently become anti-complementary when heat-inactivated at 56°C for 30 min, and generally cannot be used in standard complement-fixation (CF) assays. Therefore, a procedure was developed for decomplementing canine sera by absorption with particulate immune complexes consisting of sheep erythrocyte stroma optimally sensitized with anti-sheep erythrocyte antibody (hemolysin). Canine sera incubated for 20 min at 30°C with sensitized stroma consistently showed less than 10% residual complement and were not anti-complementary. This decomplementation procedure was applied in a complement-fixation (CF) test for detection of serum antibodies during canine visceral leishmaniasis. Two groups of German shepherd dogs were injected intravenously with Leishmania donovani or L. donovani chagasi amastigotes, and the course of infections was followed for 12 weeks. Using freeze-thaw sonicate preparations of L. donovani parasites as antigen, reciprocal CF antibody titers above 24 were detectable in sera 7 weeks after infection and gradually increased to a maximum titer of 775 at 12 weeks. Sera from control dogs had mean titers of 24. This improved methodology enhances the potential of the CF test in the serodiagnosis of canine leishmaniasis.

Accepted for publication January 27, 1984.

^* In conducting research described in this report, the investigators adhered to the Guide for Laboratory Animal Facilities and Care as promulgated by the Committee of the Guide for Laboratory Animal Resources, National Academy of Sciences, National Research Council.

The views of the authors do not purport to reflect the position of the Department of the Army or the Department of Defense (Para. 4-3, AR 360-5).

Present address: Armed Forces Institute of Pathology, Department of Automation Management Services, Washington, D.C. 20306.