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Immunoelectrodiffusion with paper discs is a simple and rapid method for screening blood donors for hepatitis antigen.

The screening of a dialysis population for the presence of hepatitis-associated antigen (H.A. antigen) has proved to be of value in locating a probable source of infection and in terminating an outbreak of hepatitis by early detection of H.A. antigen positive patients and staff.

Immunological studies were performed on Ugandan patients with hepatocellular carcinoma to test the hypothesis that the high rate of persistence of hepatitis-associated antigen in these patients is the result of defects in host immune response. The responses to 1-chloro-2,4-dinitrobenzene sensitization and to a battery of recall skin test antigens were normal, as was the humoral antibody response to tularaemia antigen. Neither hypogammaglobulinaemia nor specific immunoglobulin deficiencies were found. Thus it appears unlikely that generalized defects in host immune responses can account for the high incidence of persistent hepatitis B virus infection found in Ugandan patients with hepatocellular carcinoma.

Histopathological findings in 91 Ugandan patients with cirrhosis were studied in relation to serological tests for the hepatitis-associated antigen (H.A.A.) and antibody (anti-H.A.A.). H.A.A. was present in 30 (32·9%) of the 91 patients as opposed to 7 (3·1%) out of 224 controls drawn from the same population (P

Sera from 87 patients with acute sporadic viral hepatitis were tested for the presence of the Australia-SH antigen. Forty-three were positive by the complement-fixation test, but only 24 of these reacted in the gel-diffusion test. The antigen was equally distributed in infectious and serum hepatitis. The relationship of naturally occurring antigen-positive hepatitis to the Willow-brook MS-2 type is discussed.

Of 53 cases of active chronic liver disease two were found to be carriers of Australia antigen Au (1)-an elderly woman with typical lupoid hepatitis and an elderly mortuary attendant with serologically atypical active chronic hepatitis. Au (1) was detected also in the serum of 7 out of 20 patients with clinically atypical acute hepatitis-one was an elderly Italian woman, one had hepatitis in the puerperium, and five had a history of transfusion or inoculation. The antigen was not found in 20 typical cases of infectious hepatitis in young people in 86 patients with other diseases. Antibody to Au (1) was present in only 2 out of 102 patients who had received numerous transfusions.We conclude, firstly, that Au (1) antigen is rare in white Australians (in keeping with the low incidence of serum hepatitis in Australia), and, secondly, that Au (1) positivity in hepatitis patients is associated with transfusions and with older age. We suggest that active chronic hepatitis and lupoid hepatitis may follow infection of susceptible individuals with Au (1)-positive hepatitis virus, but persistence of the virus in high titre does not appear to be necessary for chronicity of the disease.

A 19-week-old English girl developed acute viral hepatitis, which became chronic with persistent hepatosplenomegaly and abnormal liver function tests. Liver biopsy at 1 year showed an active cirrhosis with multinucleated giant cells. The Australia (Au) antigen was detected repeatedly in the infant's serum by immunodiffusion and by electron microscopy at the time of the acute attack and during the development of cirrhosis. She had apparently acquired the hepatitis from her mother, who had had jaundice at the end of pregnancy and for one month thereafter, and who was subsequently shown to be a carrier of Au antigen. Particles with surface projections resembling paramyxoviruses were observed in two of the later specimens of the infant's serum.

In 39 unselected patients with acute infective hepatitis diagnosed on clinical, epidemiological, and pathological findings, smooth muscle (S.M.) antibody was found in 87% and hepatitis-associated antigen in 33%. The incidence of S.M. antibody was highest during the first month after the onset of symptoms, and the antibody was preponderantly IgM at this stage. S.M. antibody and hepatitis-associated antigen occurred independently, and unlike hepatitis-associated antigen, which was strongly associated with possible parenteral exposure to infection, the incidence of S.M. antibody was not related to the presumed mode of exposure. It is concluded that production of S.M. antibody is more directly related to liver cell damage than to the presence of hepatitis-associated antigen in the serum.

Examination of serum specimens from a patient with chronic active hepatitis proved negative to the Australia-SH antigen by immunodiffusion and by complement fixation. Because the sera were anticomplementary they were examined with the electron microscope, and virus-like structures similar to members of the coronavirus group were identified. The possible significance of this finding in human serum and its relation to mouse hepatitis virus are discussed.A sample of the serum pool containing the MS-1 agent was found negative when examined by immunodiffusion, complement fixation, and electron microscopy. The possible lack of immunological identity between infectious and serum hepatitis is discussed.

The sera of 110 patients with chronic hepatitis and adequate controls were examined for antibodies to smooth muscle (S.M.), mitochondria (M.), and for antinuclear factors by the immunofluorescence method, and for Australia (Au(1)) antigen by a modified micro-Ouchterlony immunodiffusion technique. Twelve out of 13 patients with primary biliary cirrhosis had M. antibodies, two had antinuclear factor, and none had Au(1) in their sera. In chronic aggressive hepatitis 23·5% of the sera contained antinuclear factor, 13% S.M. antibodies, 10·5% M. antibodies, and 22% Au(1) antigen. Of the 12 patients with chronic persistent hepatitis, one had antinuclear factor, one S.M. antibodies, and three Au(1) antigen. The most striking finding was a mutual exclusion between Au(1) antigen and M. and S.M. antibodies. None of the 33 patients with one or the other form of chronic hepatitis and M. or S.M. antibodies had Au(1) antigen; 22 out of 77 (28%) patients without such antibodies were positive.

Viral hepatitis in young adults in Accra, Ghana, is associated with Australia antigen (H.A.A.). Sera from 85 patients in hospital with viral hepatitis were available for determinations of H.A.A. Of the 16 patients whose serum was obtained within the first week of symptoms, 15 were positive. The only factor related to finding H.A.A. was the time between onset of symptoms and the collection of the serum sample. Persistence of H.A.A. was associated with persistence of jaundice in men but not in women. Previous epidemiological studies in Accra found no evidence for parenteral transmission of viral hepatitis and showed a shanty-town predilection pointing to faecal-oral transmission. It thus seems that H.A.A.-associated hepatitis is transmitted in West Africa either faecal-orally or by shanty-town associated arthropods. The finding that H.A.A. hepatitis is the usual hepatitis in young adults in Accra is in accord with the high prevalence of H.A.A. elsewhere in the general population in Africa and may be related to the high rate of cirrhosis and hepatoma in Africa.

In a comparison of 347 chronic carriers of the SH antigen and 8,864 controls with respect to ABO and Rh blood group distribution no significant differences have been detected.

During a period of one year all of 105,724 blood donations were tested for Australia (Au) antigen and its antibody by rapid immunoelectro-osmophoresis-86 (1 in 1,229) were positive for antigen and 67 (1 in 1,578) positive for antibody. Second donations by previously negative donors reduce the overall incidence of positives. Men prisoners have a significantly higher incidence of Au antigen (1 in 153) than non-institutionalized men (1 in 803). The latter have a significantly higher incidence of antigen than women (1 in 2,019). Only one antigen-positive donor was incubating acute viral hepatitis. Failure to detect one strong and one weak antigen was responsible for two cases of posttransfusion Au-antigen-positive hepatitis.

Hepatitis associated antigen may be detected quickly and reliably by the latex agglutination test, using antiserum from guinea pigs immunized with the antigen. The latex test has a sensitivity comparable to the counter current immunoelectrophoresis technique.

One of the new insights emerging through careful and imaginative study of the hepatitis B surface antigen (HBs Ag, Australia antigen) in various clinical settings is that the transmission of viral hepatitis is not fully defined. This lack of understanding contributes to serious problems, especially with nosocomial infection. In hepatitis B virus (HBV) infection, the high frequency of HBs antigenemia anti-HBs in medical personnel has repeatedly been documented, particularly in hemodialysis units.1 , 2 Now, the paper of Wands et al. in this issue of the Journal adds another disturbing dimension to the problem. In a study of HBV . . .

In collaboration with Horst in 1970 my colleagues and I reported Australia antigen in urine and feces of patients with Au-positive hepatitis, 1,2 an observation confirmed by others. Grob and Jemelka 3 found Australia antigen in the stools of patients with acute hepatitis, and Blainey et al 4 isolated it from urine of patients with renal transplants. After examination of urine and feces specimens by electron microscopy, we found not only the common 200-Angstrom particles but also the large (Dane's) particles of about 450 A diameter, which is believed to be the hepatitis virus per se. 5 Materials and Methods Serial samples of urine and feces of five patients with acute hepatitis, two with chronic hepatitis, one with subactue hepatitis, and one with Kimmelstiel-Wilson's syndrome, as well as materials from three normal controls, were collected and examined within 24 hours according to the method described later in this paper.

The duration of incubation for icteric viral hepatitis appears to be shorter for fresh blood, serum, and plasma than for stored serum or plasma, pooled or unpooled. More than half of the cases of serum hepatitis from blood occur prior to the 60th day and three fourths before the 90th day. It has become evident also that one may extend the incubation period for infectious serum and plasma almost at will, especially by the use of ultraviolet light or by storage of liquid plasma or serum at temperatures between 20 and 35 C for three months or longer. Eventually, these extensions of icteric serum hepatitis reach "infinity," in that no cases of serum hepatitis are observed. It is the purpose of this paper to present data from several sources in support of these statements and to discuss their possible relationship to the Australia antigen. In general, the longer the duration

An effective and relatively simple method for production of monospecific heterologous antisera to hepatitis-associated antigen (HAA) is described. The inoculum was prepared by concentration of pepsin-treated HAA from pooled HAA-positive sera and used to hyperimmunize rabbits and guinea pigs by a multiple-route technique. Although guinea pigs had a stronger immune response to the inoculated HAA, antisera from rabbits were more satisfactory for use in electrophoresis and immunodiffusion. This paper presents a relatively simple and efficient method for producing monospecific heterologous antiserum to hepatitis-associated antigen (HAA). The method is simpler than methods previously described, in that an effective inoculum is prepared by concentration of HAA from HAA-positive sera treated with pepsin [1-3]. The reactivities of the antisera obtained compare favorably with those produced with purified concentrates of HAA or with human antisera to HAA. Heterologous antisera to HAA were produced by various schemes of inoculation in rabbits and guinea pigs for comparison of their reactivities and specificities.

"Australia" antigen has been shown to be closely associated with serum hepatitis. The presence of the antigen and its corresponding antiserum can be detected in human beings (and in certain primates) by a number of laboratory tests. This is of great potential importance to blood transfusion and similar services because detection and exclusion of blood donors carrying the antigen might significantly reduce the risk of hepatitis from transfusions and other procedures.In this paper the present state of knowledge of "Australia" or "hepatitis-associated" antigen is reviewed. The currently employed tests are described in detail and their use, interpretation and limitations are discussed. Though it appears from early studies that the application of routine screening tests to blood donors would only reduce the risk to recipients by less than 25%, the more sensitive tests becoming available may increase this percentage and it is recommended that where competent laboratory services are available steps should be taken to set up a scheme for testing donors-provided that the current limitations of such a scheme are clearly recognized.

Light and electron microscopic examinations were carried out on liver tissues from 6 renal transplant recipients who developed Australia antigen-associated hepatitis, referred to in this paper as hepatitis-associated antigen (HAA) hepatitis. While under immunosuppressive therapy, the patients showed rapid progression of acute hepatitis to chronic active hepatitis and posthepatitic cirrhosis resulting in liver failure. By electron microscopy, in 5 of 6 patients, characteristic virus-like particles of 230 Å size were observed in massive numbers in the nucleus and occasionally in the cytoplasm of liver cells. No similar particles were found in liver biopsies from other patients. These particles had an appearance of nucleocapsid without an outer coat, and were believed to be the viruses in their natural infective states. The predominantly intranuclear location of the particles and the associated nuclear changes suggested that the nucleus is the primary site of virus replication. The finding of the virus-like particles is believed to be diagnostic for HAA hepatitis. While HAA is implicated in serum hepatitis, the identification of HAA with the virus-like particles of this study is still to be established. In the phagosomes, there were some particles showing an outer membrane and a tail-like structure. Both the coated and the noncoated particles were believed to be the same virus. The phagosomes also contained what appeared to be “degradated” viral material. The lysosomal activity in the infected hepatitic cells seemed to be increased. This study showed nonspecific changes of the liver cell nuclei and organelles, probably related to viral infection. The precise mechanism of cell injury remains to be elucidated. However, the result of this study has provided a reasonable basis for future investigations of HAA hepatitis.

The platelet aggregation test (PLA) can be used as an indicator of Australia antigen or antibody in the serum. 1,2 The reaction of Au antigen-Au antibody complexes with human platelets changes the pattern of sedimentation of platelets on microplates. 2 This method has also been used to detect other antibodies and antigens. 3 When PLA was used to study hepatitis sera, it was soon found that many of these sera reacted directly with platelets without any added antibody or antigen. Earlier studies show that this can be a sign of the presence of immune complexes in the blood. 4 The possibility of immune complexes in severe hepatitis cases has recently been under special consideration. 5 This paper presents the results of 93 cases of different forms of hepatitis, 49 other diseases, and 99 blood donors. Special consideration is given to the "direct" reactivity of sera. The detection of the