Preparation of noninfectious hepatitis A virus hemagglutinin for detecting hemagglutination inhibition antibodies.
Hepatitis A virus (HAV) harvested from infected MRC-5 cells can hemagglutinate various species of erythrocytes at acid pH (Eckels et al., 1989). Further studies revealed that the majority of the hemagglutinin (HA) in MRC-5 and BS-C-1 cells was cell-associated. A simplified procedure for preparing HAV-HA consisted of collecting infected cells in phosphate-buffered saline followed by three cycles of freeze-thawing and sonication. The fluids were clarified and stored at 4 degrees C. The analysis of HA by rate-zonal sucrose gradient centrifugation indicated that the majority of HA co-migrated with infectious virus. Complete inactivation of infectious HAV with 0.03% beta-propiolactone (BPL) did not affect HA activity, while inactivation with 0.05% formalin caused a 16-fold reduction in titer. There was no difference in HAI antibody titers when BPL-treated HA was compared to untreated HA in the hemagglutination inhibition (HAI) test.
Dubois
D R
DR
Division of Communicable Disease and Immunology, Walter Reed Army Institute of Research, Washington, DC 20307-5100.
Binn
L N
LN
Summers
P L
PL
Timchak
R L
RL
Barvir
D A
DA
Marchwicki
R H
RH
Eckels
K H
KH
D000818:Animals; D002478:Cells, Cultured; D002498:Centrifugation; D006385:Hemagglutination Inhibition Tests; D006389:Hemagglutinins, Viral / Q000378:metabolism*; D006508:Hepatitis Antibodies / Q000032:analysis*; D006507:Hepatovirus / Q000276:immunology*; D006801:Humans; D011420:Propiolactone / Q000494:pharmacology; D014775:Virus Activation / Q000187:drug effects
D006389:Hemagglutinins, Viral; D006508:Hepatitis Antibodies; D011420:Propiolactone