Alfred M. Prince

Serum Hepatitis Antigen (SH): Rapid Detection by High Voltage Immunoelectroosmophoresis

An immunoelectroosmophoretic technique for rapid detection of the antigen (SH) associated with the serum hepatitis virus has been devised. The technique maintains the specificity characteristic of the Ouchterlony gel-diffusion method, yet detects in 1 to 2 hours one-tenth the amount of antigen required for gel diffusion. The test has immediate application to blood-banking practice since it permits the screening of such labile products as platelets and fresh whole blood, and the detection of antigen in additional serums negative by the Ouchterlony technique.

Role of immune complexes involving SH antigen in pathogenesis of chronic active hepatitis and polyarteritis nodosa.

Abstract The passive haemagglutination assay has revealed circulating SH antibody in the serum of 16 of 58 chronic SH carriers. The quantity of this antibody did not correlate well with the degree of liver damage. Rate-zonal density-gradient centrifugation revealed sera with coexisting antigen and antibody to contain rapidly sedimenting immune complexes. The presence of such complexes did not correlate well with the presence or absence of chronic active hepatitis or polyarteritis nodosa. These and other data are interpreted as suggesting that, although the pathogenesis of chronic hepatitis and polyarteritis nodosa most probably involves immune mechanisms, these are not mediated by pathogenic immune complexes composed of SH antigen and antibody.

HEPATITIS B ANTIGEN IN WILD-CAUGHT MOSQUITOES IN AFRICA

Abstract 187 pools of mosquitoes caught in the wild in Kenya and Uganda have been examined for the presence of the hepatitis B (H.A.A., SH, Australia) antigen. This antigen was detected by solid-phase radioimmunoassay in 28 pools representing eight different species. These data suggest a possible role for blood-sucking arthropods in transmission of hepatitis-B infections in the tropics and in other areas. The high prevalence of chronic hepatitis-B infections in the tropics may depend on blood-sucking arthropod vectors to assure a high frequency of exposure in the early years of life when the risk of developing chronic hepatitis-B-virus infections is greatest.

The high voltage immunoelectroosmophoretic (IEOP) technique for detection of SH antigen: application to blood donor screening and to study of liver disease.

An assay for the serum hepatitis antigen (SH/Australia/hepatitis associated antigen) which can utilize presently available impure reagents and is able to provide results with sufficient rapidity to permit screening of labile blood products has recently been reported [PRINCE and BURKE, 19701. This procedure, which we have termed high voltage immunoelectroosmophoresis (IEOP), is a modification of the counterelectrophoresis method in which antigen is caused to migrate in an electric field through a layer of agarose against antibody migrating in the opposite direction due to endosmotic flow. Similar methods of screening for SH antigen, which utilize a lower voltage, have recently been described [PESENDORFER et al., 1970; GOCKE and HOWE, 19701. Comparative assays carried out on 12 different SH antigen containing sera revealed the IEOP technique t o be capable of detecting on the average ten-fold less antigen than the standard agar gel diffusion assay. A similar estimate of sensitivity has been reported by GOCKE and HOWE [1970]. The present report summarizes comparative results obtained using the IEOP and the Ouchterlony technique t o screen sera from clinically well individuals, and from patients with acute and chronic liver disease for SH antigen. In order to evaluate the sensitivity of the IEOP test for detection of SH antigen, 1,023 sera from different populations with a known high frequency of antigen carriers were tested by the IEOP and agar gel diffusion techniques (table I). I n

Kinetic Studies on the Direct Solid Phase Radioimmunoassay for Hepatitis B Antigen

Abstract. Kinetic studies have revealed the direct solid phase radioimmunoassay for hepatitis B antigen (Ausria, Abbott Laboratories) to be markedly sensitive to incubation temperature. The present 18‐hour room temperature incubation can be reduced to 2 h at 50 °C without loss in test sensitivity. The rapid test is also considerably more specific than that carried out under standard conditions.

Recent Advances in our Understanding of Hepatitis B Infections

Abstract. The development of highly sensitive hemagglutination (HA) assays for detection of hepatitis B antibody has made possible observation of primary anamnestic antibody responses in most individuals exposed to hepatitis B virus and the development and evaluation of a hyperimmune γ‐globulin preparation.

Detection of Serum Hepatitis Virus Carriers by Testing for the SH (Australia) Antigen

In 1964 BLUMBERG described what appeared to be another human serum protein polymorphism: an antigen detected in the serum of an Australian aborigine which was found to react in the Ouchterlony technique with an antibody derived from the serum of a multiply transfused hemophiliac [l]. This antigen, which BLUMBERG termed the Australia antigen, was detected in a proportion of sera from some foreign populations and from patients with leukemia [2]. Family studies appeared to support the hypothesis that this antigen was a genetically determined isoantigen [3]. The Australia antigen was subsequently shown to be present in the sera of about 25% of institutionalized patients with Down’s syndrome. Serial specimens drawn from one child with Down’s syndrome did not reveal Australia antigen until shortly before the onset of hepatitis in this patient. Additional testing revealed antigen in 5 out of 48 patients with viral hepatitis [4].

CAN THE BLOOD-TRANSMITTED HEPATITIS PROBLEM BE SOLVED?

Of the approaches presently available for prevention of blood-transmitted hepatitis, most clearly effective is the avoidance of high-risk donor populations, such as most paid commercial donors. It should be emphasized that even after screening for hepatitis B antigen most paid donor blood carries with it a five-ten times higher risk of transmitting hepatitis than volunteer donor blood. Hepatitis B screening is certainly the next most useful approach. However, it has been pointed out that even with the most sensitive of current assays, as much as two-thirds of infective blood may not be eliminated. In part this is due to failure to detect hepatitis B antige, which may be detectable by such currently investigational approaches as selective antibody inhibition or testing for Core antibody. However, a major factor is the inability of hepatitis B screening to eliminate all infectious blood appears to be due to the existence of viruses other than hepatitis B that play a major role in blood-transmitted hepatitis. Tests for detection of carriers of this virus, or viruses, remain to be developed. Improved reporting of hepatitis cases with investigation and registration of implicated donors may provide an additional means for reducing the incidence of hepatitis transmission. The efficiency of additonal approaches, such as transaminase screening of donors and passive or active immunization, remains to be determined.

ELECTRON MICROSCOPIC OBSERVATIONS ON SH ANTIGEN USING POSITIVE STAINING

This chapter presents the electron microscopic observations on serum hepatitis (SH) antigen, which have been obtained using positive staining. SH antigen, which is identical with the Australia (Au) antigen, has been observed in the serum in acute and chronic liver diseases. Electron microscopy shows that this antigen is located on a 200 A spherical structure. The results of the experiments indicate that the outer membrane of the 430 A particles is similar to the membrane of the 200 A particles, in staining characteristics and morphology. Because only the central core of the large particle stains positively with uranyl acetate, only this form of SH containing particle can contain nucleic acid and possibly be a virus. The electron microscopic observations, based on differences in staining properties, indicate that only the larger of these particles might contain nucleic acid.