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Half the Dane particle concentrates isolated from 84 serum samples of patients infected with hepatitis B virus (HBV) contained more than one HBV DNA species. At most, six HBV DNA molecules migrated on hybridization blots. The fastest migrating species was single minus-strand DNA, and the five slower species all were double-stranded relaxed circular DNA. One of the latter forms corresponded to the full-length HBV genome; the four others contained shorter plus strands of different lengths and might represent replicative intermediates. These results indicate that the replicative forms of HBV previously found in infected liver cells might be coated and exported as early as the step of single minus-strand synthesis.

In a 12 year-old boy presenting with auto-immune hemolytic anemia of the IgG type, human parvovirus infection was responsible for acute erythroblastopenia. Aplastic crisis quickly and spontaneously recovered but auto-hemolysis was durable. Human parvovirus induced erythroblastopenia has only been reported in patients with constitutional hemolytic anemia. On the other hand, in this case, human parvovirus infection revealed the auto-immune hemolytic anemia.

One hundred and thirty six patients receiving haemodialysis in a HB antigen-free unit were prospectively studied over a period of 29 months for evidence of hepatitis. Twelve/one hundred and eleven patients who were dialysed in this unit for at least one month developed elevation of ALT which proved to be related to neither toxic hepatitis nor hepatitis due to any of the following viruses: hepatitis B virus (HBV), hepatitis A virus (HAV), cytomegalovirus (CMV) or Epstein-Barr virus (EBV). Therefore these cases were considered to be non-A non-B (NANB) hepatitis. In 5 patients the liver disease was of short duration, whereas in 7 others hepatitis had a chronic course with ALT remaining elevated for more than 6 months. During the same period, one/sixty staff members who were working for at least one month in this unit also developed presumed non-A non-B hepatitis. Serological markers of NANB infection tested by double immunodiffusion were present in 10/12 patients and in the one staff member.

HTLV infection was detected in normal French blood donors and haemophiliac patients using HTLV p24 radio-immunoassay. No positive was found in 772 sera from continental blood donors born in continental France or in foreign countries. Among 131 donors living in continental France and born in overseas territories, 2 (1.5%) were found to possess anti-HTLV p24 antibodies. Similarly in a sample of 1 239 donors collected in Martinique, 1.5% were positive. No positive had been found in a sample of 142 continental haemophiliacs.

Quantitative analysis of beta-2-microglobulin (beta-2-M) may be a useful test for early diagnosis of acquired immuno-deficiency syndrome (AIDS). Even if beta-2-M is an unspecific marker, 96% of 45 patients with confirmed AIDS and 82% of 45 patients with lymphadenopathy have excessive (3 mg/l) beta-2-M value. beta-2-M is also useful in epidemiological studies, especially in subjects in close contact with patients, and in polytransfused hemophiliacs. At last, beta-2-M may be useful as a screening method in blood banks: from a study of 929 sera, excessive beta-2-M value was observed in 16 cases, which were excluded as blood donors.

From January 1980 to December 1983, 989,907 blood donations were screened for HBsAg. A total of 1,345 was found HBsAg positive; 33 of whom were negative for anti-HBc (2.45%); in 10 cases HBeAg was present at a low level along with HBsAg. 2 of the 33 subjects were lost to follow-up. Late serum samples were available for 28 blood donors, and clinical and biological data only were known for the 3 others. An evolution of HBV markers was observed in each case; anti-HBc antibody became positive in 24 subjects (the 4 remaining subjects had a very short follow-up); HBeAg was positive at a high titer in 18 subjects and seroconversion to anti-HBe was observed in 7 individuals; HBsAg concentration increased in 21 subjects from 2 to 4,000 times and decreased in the 7 others. Seroconversion to anti-HBs was observed in 8 individuals. According to these serological, biological and clinical findings, different outcomes were observed: in 6 subjects too short a follow-up was available to allow appropriate classification, but there was evidence to suggest all probably developed clinical hepatitis; in 6 subjects asymptomatic hepatitis with rapid loss of HBsAg occurred; in 2 subjects asymptomatic hepatitis with the persistence of HBsAg at a low level and seroconversion to anti-HBe occurred; and in 17 subjects clinical hepatitis developed. These observations establish that most of the 33 blood donors were infectious at the time of their blood donation, and that anti-HBc antibody screening must not replace HBsAg screening for blood donations.

Forty nine blood donors immune to HBV volunteered to receive a single dose of hepatitis B vaccine (HEVAC B from Institut Pasteur Production). The initial mean level of anti-HBs was 20 IU/ml (from 5 to 75 IU/ml). The maximum level was observed at the consecutive plasma donation usually 1 month and a half after the vaccine injection. The mean maximum level was 188 IU/ml (from 7 to 800 IU/ml) i.e. an increase of 9 times the initial level. Six donors developed a very weak immune response (inferior to twice the initial level). In the 43 others, anti-HBs levels remained higher than initial level 6 months after the vaccine injection. One year later (study conducted in 25 subjects) 56% had anti-HBs levels superior to 3.6 to 16 times the original level. Such booster responses are of considerable practical importance for increasing anti-HBs titers in human donors for producing HBIG.

A survey conducted in sera from 515 black and 499 white individuals living in 8 villages of Mauritania showed an HBs Ag prevalence of 15.7%. Two main subtypes, ayw2 (34.7%) and ayw4 (63%), were found in this country which is a junction area between North and West Africa and where 2 ethnic groups are living close one to the other but with very rare cross-breeding. ayw2 was more prevalent in the driest part of the country (North) and ayw4 in the South. Analysis of the subtype distribution in each village indicates that there is no relationship between HBs Ag subtype and ethnic groups, but there is a correlation between HBs Ag subtype and the geographical location of the villages.

From March 1984 through November 1985, 24 children and adults with aplastic crises were admitted in several Parisian hospitals. Twelve patients had known hemolytic anemia. Aplastic crisis revealed hemolytic anemia in the remaining patients. The detection of human parvovirus antigen was performed by counter-immunoelectrophoresis, and specific IgM antibodies were detected by IgM-antibody-capture-radioimmunoassay, in order to establish the incidence of human parvovirus infection in the genesis of the aplastic crisis. Twenty-one patients had acute infection with human parvovirus. In the three remaining patients, no marker of human parvovirus infection was found. The features of the human parvovirus linked aplastic crisis are described. We consider that human parvovirus infection, and unknown hemolytic anemia, must be systematically researched in any case of unexplained acute aplastic anemia.

HHV.6 (or HBLV) herpesvirus was isolated in 1986. This virus has a specific tropism for T cells. It is ubiquitous. Most subjects are infected in early childhood. HHV.6 is responsible for exanthema subitum. Its possible involvement in the aggravation of immunodeficiency syndromes and its action on malignant haematologic disorders, has been suggested but has not been clearly demonstrated.

Seven hundred and forty subjects who, between 1980 and 1985, had received intravenous or intramuscular injections of immunoglobulins (Ig) prepared by Cohn fractionation of plasma pools from more than 2000 donors were investigated for anti-HIV antibodies. Anti-HIV antibodies were detected in only one subject, a female drug addict and therefore belonging to a group at high risk of AIDS. In contrast with the huge amounts of Ig received by the remaining 739 subjects (976 litres intravenously, 128 litres intramuscularly), this young woman had only received 10 ml of anti-HBs Ig intramuscularly. The wide scattering in both space and time of these recipients of Ig and the prevalence of anti-HIV antibodies among blood donors in France (0.7 per 1000 in 1985) suggest that all the batches studied (over 100) were contaminated by HIV. The absence in these subjects of seropositive reaction ascribable to the injections of Ig confirms that Cohn fractionation inactivates the virus, as demonstrated in vitro. It is concluded that recipients of Ig prepared from plasma pools not tested for the absence of anti-HIV antibodies should not be regarded as a group at risk of AIDS.

A serum panel obtained from male homosexuals (n = 278); i.v. drug abusers (n = 99), patients attending a VD clinic (n = 390), blood donors who visited Central or West Africa (n = 573), blood donors who had sexual contact with natives from Central or West Africa (n = 38), blood donors from Surinam, South America (n = 481), individuals positive for anti-HIV-1 (n = 94), individuals with indeterminate HIV-1 western blot (WB) reactions (n = 73), African patients with AIDS or AIDS-related symptoms (n = 30), and random Dutch blood donors (n = 555), was tested with HIV-2 ELISA (ELAVIA-2). Of these 2,611 samples, 32 (1.2%) were repeatedly reactive. Antibodies to gp140/105env were found in 4/32 by HIV-2 WB, and in 1/4 by radioimmunoprecipitation (RIPA). These 4 HIV-2 WB-positive samples were also reactive with gp160/120env in HIV-1 WB, suggesting cross-reactivity. In a spot test with synthetic peptides of the transmembrane glycoprotein of both HIV types, 3/4 were only HIV-1 positive and 1/4 was strongly HIV-2 positive and weakly HIV-1 positive. In inhibition assays with soluble HIV-1 or HIV-2 synthetic peptides in HIV-1 and HIV-2 peptide ELISA, cross-reactivity was excluded, which indicates an HIV variant or HIV-1/HIV-2 double infection. It is concluded that for the moment HIV-2 infection is at low prevalence in risk groups in The Netherlands, and that in addition to WB and RIPA, synthetic peptide assays are useful for differentiation between HIV-1 and HIV-2 antibodies.

212 male prisoners were collected at the prison in March 1983. Anti-HBc and HBs Ag were detected by a combined test (AUSRIA-CORE, Abbott Laboratories). 67 sera (31,5%) were anti-HBc positive, 7 of them HBs Ag positive. Screening for anti-HTLV (anti-p24) was negative for all the donors. Beta 2 microglobulin levels were determined on 115 sera (B2-micro RIA 100, Pharmacia Laboratories). 8 had levels above 2,6 mg/l (greater than 2 SD). These 8 sera were anti-HBc positive and one of them HBs Ag positive 3 HBs Ag positive donors had non elevated Beta 2 microglobulin levels. This survey confirms that prisoners as blood donors should be regarded as carrying a high risk of transmission of HBV and probably other infectious agents with similar epidemiology. The signification of elevated Beta 2 microglobulin levels deserves further investigations since this determination could be of value as an additive test to increase the safety of blood products.

A seroimmunologic evaluation of 57 children with chronic hepatitis is presented. Twenty-one patients had chronic persistent hepatitis and 36 had chronic active hepatitis. Serum samples obtained before treatment were tested for HBsAg, anti-HBs, anti-HBc, smooth muscle antibody, and antibody to endoplasmic reticulum. A persistently positive HBsAg was observed in the serum of 18 of the 21 patients with chronic persistent hepatitis. The chronic active hepatitis group was divided into three subgroups according to the presence of hepatitis B-virus markers (7 patients), smooth muscle antibody (10 patients), and endoplasmic reticulum antibody (9 patients). Determination of these markers could be useful for classifying children with chronic hepatitis.

Spleens from high responder Biozzi mice immunized with purified HBsAg of adw2, adw4 and ayw3 subtypes respectively were fused with N5-1 mouse myeloma cells. Over 30 clones with anti-alpha activity were obtained (principally from fusion adw2) as well as numerous clones with restrictive anti-HBs reactivity. The aim of this study was to characterize 9 different monoclonal antibodies with restrictive reactivity in comparison to a monoclonal anti-alpha antibody. Two different techniques were used: immunodiffusion and enzymoimmunoassay. Already known specificities or common reactivity between some subtypes were found, as w4, subdivision of ayw3 subtype, antigenic determinant common to w4 and adr, absence of the same determinant in adw2 and ayw1, the 2 subtypes having the highest capacity to absorb anti-alpha. Other monoclonal antibodies indicated the presence of previously unidentified subtype specificities: one of them recognized all HBsAg with d determinant but also ayw4, another recognized all HBsAg with y determinant but also adr, 2 others recognized determinants present completely or partially (spur formation) in many subtypes, another one recognized ayw4 and adw4 but also adw2. The combination of new tools such as monoclonal antibodies and synthetic peptides should facilitate the analysis of HBsAg.

A seroepidemiological surveillance study to investigate the spread of HIV-2 infection in France was organized in the last semester of 1987 on 100 114 blood donors and 10 004 selected people. Sera were simultaneously tested for antibodies to HIV-1 and to HIV-2 by enzyme-linked immunosorbent assay (ELISA) and were confirmed by specific Western blot. A spot test using synthetic peptides was also applied when a sample was reactive by ELISA HIV-1 and HIV-2 and was confirmed as HIV-1 by Western blot. A total of 30 blood donors were confirmed as HIV-1 positive (0.030%) and no sample was confirmed as HIV-2 positive. In the selected population, the prevalence of HIV-infected people was 7.1% for HIV-1 and 0.09% for HIV-2. Eight of the nine HIV-2 cases were directly or indirectly connected to West Africa. Only one HIV-2 case had no known contact with these countries; he has been transfused and is under investigation. Except for this possible HIV-2 infection in a blood recipient, no HIV-2 seropositivity was found in at-risk groups (homosexuals, drug addicts and haemophiliacs). At present, HIV-2 does not seem to be widely spread in France. Seroepidemiological surveillance will be continued.

This report was presented at the June 1983 meeting of the Blood Transfusion National Consultative Commission at the French Ministry of Health. Clinical and epidemiological data on AIDS as well as problems raised by the lack of specific tests for screening of blood donors were briefly summarized. Out of 49 AIDS patients recorded in France up to April 1983, only one had a history of previous blood transfusion given in Haiti, 4 years before the clinical onset of the disease. Blood donors, all Haitians, had no sign and symptom of AIDS. Retrospective review of 2 300 hemophiliacs followed up in France until April 1983 disclosed no AIDS. However, in 6 patients, the following features, more or less associated, were found to be present: thrombocytopenia, neutropenia, micropolyadenopathy, splenomegaly, hypergammaglobulinemia and low OKT4/OKT8 ratio. No clear correlation could be found between these abnormalities and the origin, commercial or national, of the coagulation factor concentrates used for the treatment. Three main recommendations were proposed: -- information of blood donors and experimental evaluation of some non specific screening tests, in the at risk donor population. -- more cautious use of coagulation factor concentrates -- reduction of importations aiming at complete national self sufficiency concerning factor VIII concentrates.