Various clinical and pathological conditions can develop after EBV infections depending on the age ofthe individual. Whereas young children show a silent seroconversion, adolescents with acute EBV infection develop infectious mononucleosis (IM) in half of the cases . Being a lymphotropic virus, EBV can trigger an unspecific B cell stimulation besides the specific immune response. This may cause a big rise of class G and M Igs in the sera. Little is known about their specificity. Heterophil antibodies appear that agglutinate sheep, horse, and bovine erythrocytes (1, 2). Cold antibody with anti-i specificity was found in 8% to 60-70% of the IM patients (3, 4). In addition, antibodies are found that react with components of the cytoskeleton (5) or possess antinuclear features (6). None of these antibodies always appears in acute EBV infection, thus, their significance for the pathogenesis of IM is not clear. This may be different in the antibodies we recently detected in all sera of persons acutely infected with EBV These antibodies possess specificity for the glycolytic enzyme, triosephosphate isomerase (TPI).
Volume 171 February 1990 565-570
Materials and Methods
Brief Definitive ReportPreparing the Antigens. Frozen cells (1 ml packed volume) and disrupted tissues were allowed to thaw, and were subsequently resuspended in equal volumes of modified lysis buffer (7) containing 9.5 M urea, 2% (wt/vol) NP-40, and 5% 6-ME; further extraction was done by five cycles of freezing and thawing (-20°C/room temperature). The cell lysate was then centrifuged at 3,000 g for 15 min. The supernatant fluid was used for antigen preparation by SDS-PAGE followed by immunoblotting. The protein concentration (20 mg/ml) was measured by the method of Neuhoff et al. (8) .PAGE. Composition and electrophoresis of SDS-13 % (wt/vol) polyacrylamide slab gels were performed as reported by Laemmli (9) . The electrotransfer of proteins from SDS gels to hydrophobe polyvinylilenedifluoride (PVDF) membrane (Millipore Continental Water Systems, Bedford, MA) was performed by transversal electrophoresis (10).Immunostaining the Blots. For the detection of antibodies, sera, as well as secondary antibodies conjugated with peroxidase, were diluted in 20% FCSTBS (vol/vol) . Each antibody
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