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Updated: Jun 19, 2026

An Optimized Hemagglutination Inhibition (HI) Assay to Quantify Influenza-specific Antibody Titers
Published on: December 1, 2017
THE NATURE OF NON-SPECIFIC INHIBITION OF VIRUS HEMAGGLUTINATION
W F Friedewald1, E S Miller, L R Whatley
1Department of Bacteriology and Immunology, Emory University School of Medicine, Atlanta.
Abstract:
A study of the component in serum and tissue extracts responsible for non-specific inhibition of hemagglutination with mumps virus and the PR8 and Lee strains of influenza virus has yielded the following results: 1. The inhibitory factor was found in high titer in human serum and in saline extracts of various organs procured at autopsy (lung, liver,kidney, spleen). The inhibition titers of extracts of these organs were usually higher than the serum titers, whereas the titers of muscle extracts were invariably lower. 2. Similar results were obtained with serum and tissue extracts from normal rabbits and guinea pigs. 3. The serum inhibition titers were not affected by heating to 75 degrees C. for 30 minutes, whereas the titers of the tissue extracts were usually reduced by heating at 65 degrees C. or 75 degrees C. for 30 minutes. 4. Saline extracts of human and chicken red blood cells also contained an inhibitory substance in high titer, and these cells showed marked agglutination with influenza and mumps viruses. Rabbit red cells, on the other band, underwent little or no agglutination with these viruses and extracts of these cells failed to cause inhibition. Sheep red cells varied in their capacity to agglutinate and also in their yield of the inhibitory substance. 5. When the virus receptor substance was removed from chicken red cells by adsorption and elution with influenza virus, extracts of the cells no longer yielded the inhibitory factor. 6. The inhibitory substance did not neutralize influenza virus in mice and it failed to fix complement when mixed with influenza or mumps viruses. 7. Evidence was obtained that some virus was released from the inhibitory substance after incubation for 6 hours at 22 degrees C. or 37 degrees C. The implications of these findings are discussed.
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