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Hepatitis B vaccine (Merck, Sharp & Dohm) was administered according to various vaccination schedules to 12 patients in preterminal renal failure without haemodialysis treatment, 81 patients on a chronic haemodialysis programme and 43 staff of a nephrological centre. After three-times vaccination (at 0, 1, 6 months) with 20 micrograms HBs antigen, seroconversion occurred in 91% of persons without kidney disease. A double dose (40 micrograms) was given, at the same time intervals, to 29 dialysis patients, with a seroconversion rate of 55% and titre levels less than in those without renal disease. Of 26 dialysis patients given the same dose (40 micrograms) five times (at 0, 1, 2, 4, 6 months), 63% achieved seroconversion with a definitely higher anti-HBs titre than those dialysis patients vaccinated only three times. Passive-active vaccination with 40 micrograms HBs antigen (at 0, 1, 2, 4, 6 months) as well as 3 ml hepatitis B immunoglobulin (at 0, 2 months) was given to 26 dialysis patients. There was a comparable frequency of seroconversion (58%) and a comparable titre rise to those obtained with active vaccination alone. Twelve pre-uraemic patients not requiring dialysis (serum creatinine 753 +/- 184 mumol/l) also were actively vaccinated three times, the results not differing from those in dialysis patients. The findings indicate that patients with renal disease potentially needing dialysis should, in the early stages of renal failure (serum creatinine less than 500 mumol/l) be vaccinated with hepatitis B vaccine.

In 195 patients vaccinated against hepatitis B the course of anti-HBs concentration was followed over 4 years. Persistence of anti-HBs proved to be dependent on the level of anti-HBs concentration after basal immunization: in 14 subjects with a maximal anti-HBs level between 10 and 100 IU/l the level had dropped to less than 10 IU/l (considered to be the lowest prophylactic concentration), while 7 were anti-HBs negative. Of those who had 101-1000 IU/l after initial immunization 49% had anti-HBs levels under 10 IU/l after 4 years, while 18% were negative. Among subjects with concentrations 1001-10 000 IU/l after the third immunization only 7% had values below 10 IU/l after 4 years, 4.2% were negative. All those who, after the third immunization, had had anti-HBs levels above 10 000 IU/l, 4 years later still had anti-HBs levels of more than 100 IU/l (mean 581 IU/l). Quantitative anti-HBs determination after triple vaccination against hepatitis B thus makes it possible to predict the duration of protection and to determine the timing of re-vaccination.

Sera obtained from 927 drug addicts in 1983 to 1985 were tested for antibodies against LAV/HTLV-III. There was a steadily rising proportion of positive results: 10.1% in 1983, 17.6% in 1984 and 23.9% in 1985. In each year the prevalence of anti-LAV/HTLV-III was higher among female than male addicts. No increased proportion of positive results was demonstrable in relation to age. Among 152 sera from 1983/84, hepatitis-B markers were found in 72 (43.7%), of whom 10 (14%) were also anti-LAV/HTLV-III positive. Among hepatitis-B marker-negative sera there were 8 (6%) which were also anti-LAV/HTLV-III positive. The prevalence of anti-LAV/HTLV-III in drug addicts in prisons, rehabilitation centres, hospitals and medical practices was similar. There is a danger that prostitution by addicts for obtaining drugs will cause a penetration of LAV/HTLV-III in the rest of the population.

Anti-B19-IgG antibodies were tested for by the ELISA method in 768 sera, 76 from children and juveniles, aged 1-15 years, attending the Outpatients Department of the Children's Clinic, University of Munich, and 692 from persons, aged 18-68 years, attending the blood donor service of the Bavarian Red Cross in Munich. 38.4% of sera were positive, with a significant difference between men and women (32.5% vs. 47.5%, P less than or equal to 0.01). A fresh B19 infection was present in 42 subjects by demonstrating anti-B19-IgM in the ELISA test. The sera came from seven patients with infectious erythema, 26 with skin rash of uncertain cause, two with erythroblastosis fetalis, two with enteritis, one with transitory anaemia, one with juvenile polyarthritis, one with lymphadenitis and arthralgia, and two with an asymptomatic course. Viral B19-DNA was demonstrated by nucleic acid hybridization in a blood unit from an asymptomatic blood donor and in amniotic fluid in a case of erythroblastosis fetalis.

One to three repeat vaccinations were undertaken in 63 persons (32 males, 31 females; mean age 29 [17-59] years) who had not or inadequately responded to basal immunization with hepatitis B vaccine (47 non- and 16 hyporesponders). After re-vaccination, 12 of the 47 nonresponders and 11 of 16 hyporesponders formed specific antibodies against hepatitis B surface antigen, anti-HBs, at concentrations above 10 IU/l. Six of 14 persons who had not responded to the first repeat vaccination reached anti-HBs concentrations above 10 U/l after a further injection. Vaccination response was age-dependent. The median age of those subjects who responded to the first re-vaccination was 27 (19-45) years, while the non-responders' median age was 34 (22-59) years (P less than 0.05). Re-vaccination also raised initially low anti-HBs values. These results indicate that some of the nonresponders can obtain reliable protection by revaccination(s).

By routine screening for serologic markers of hepatitis A and B in patients with acute hepatitis, 30 chronic carriers of hepatitis B virus with serologic evidence of acute hepatitis A and two patients with simultaneous acute infection with hepatitis A virus and hepatitis B virus were detected. For evaluation of clinical data, two major risk groups were distinguished. Nine patients were drug addicts and 17 were children and young adults from Mediterranean countries or southeast Asia. During the acute phase of illness, serum bilirubin and SGPT levels did not differ from those in other patients with acute hepatitis A. In three patients for whom follow-up sera were available, HBsAg concentration decreased during the acute stage of hepatitis A.

Cloned antibodies to specific epitopes of hepatitis D virus were produced by transformation with Epstein-Barr virus and subsequent cloning of peripheral blood B lymphocytes from a patient with chronic hepatitis D virus infection. Several stable cloned B cell lines, derived from two parent cultures, produced hepatitis D-virus-specific IgG antibodies. Some cloned IgG antibodies detected hepatitis D virus-associated antigen in hepatitis D virus-infected woodchuck liver tissue sections by indirect immunofluorescence staining and some reacted in an inhibition ELISA test detecting hepatitis D virus antibodies; most cloned IgG lines detected hepatitis D antigen both in immunofluorescence tests and in inhibition ELISA. Cloned antibodies to hepatitis D antigen detected by ELISA and/or immunofluorescence staining recognized the two major specific native and denatured polypeptides, p27 and p29, in Western blot analysis. Such cloned antibodies for hepatitis D virus are potentially useful for clinical diagnosis and research.

Before hepatitis A virus (HAV) was identified, spread of hepatitis A was prevented by public health measures. The first specific, preventive measure for hepatitis A was passive protection with standard, pooled human immune globulins. Human immune globulin contained sufficient HAV neutralizing antibodies for short-term, prophylactic passive protection and for control of the spread of local outbreaks. After many unsuccessful attempts, HAV was propagated in cell cultures and the development of vaccines for active immunization began. Formalin-inactivated, whole HAV induced protective immunity, and such formalin-inactivated hepatitis A vaccines are now being evaluated in large-scale clinical trials. HAV attenuated by serial propagation in cell culture has been used for several, live, attenuated hepatitis A vaccines and results of clinical trials are reassuring. Future approaches to protection against hepatitis A are likely to include vaccination with: hybrid viruses; hepatitis A antigen-expressing, genetically-engineered bacteria; purified hepatitis A antigens produced by molecular biological techniques and incorporated into slow or pulse-releasing systems; synthetic peptides or idiotypes.

The occurrence of antibodies to hepatitis C virus (HCV) was investigated in 81 patients who developed hepatitis non-A, non-B (HNANB) after parenteral administration of contaminated immunoglobulin to prevent Rh sensitization. Sera from 74 of the 81 patients (89.9%) were anti-HCV positive at either 6–12 months or 9–10 years after administration of immunoglobulin. Sera were not available from any patients at either of the times however, 52 of 56 sera (92.9%) were anti-HCV positive 6–12 months after use of immunoglobulin, and anti-HCV was present in 45 of 65 sera (69.2%) 9–10 years after immunoglobulin treatment. Of the latter, only two of 13 (15.4%) sera from patients who recovered from hepatitis were anti-HCV positive, whereas 43 of 52 patients (82.7%) with chronic disease were anti-HCV positive. The ELISA using a recombinant antigen was found a good detector as marker for a HCV infection because 90% of patients infected by a common source became anti-HCV positive. However, 10 years after infection most patients who did not develop chronic disease no longer had detectable antibodies.

Parvovirus B19 exerts a highly selective cytopathic effect on erythroid progenitor cells. Studies so far on the pathogenesis of B19-infection have been performed using bone marrow samples providing large amounts of erythroid progenitor cells. Extensive study, however, has been hampered by the limited access to bone marrow samples. We have designed a liquid culture method allowing the generation of large numbers of erythroid progenitor cells, initiating cultures with CD3- and CD14-poor peripheral blood mononuclear cells. Following a 12 d preincubation in liquid cultures containing recombinant human interleukin 3 (rhIl-3) and recombinant human erythropoietin (rhEpo), cells harvested from the liquid cultures were exposed to B19-containing plasma, followed by a further cultivation in liquid culture for up to 96 h. Cells expressing the CD13 and the glycophorin A (GlyA) antigens, respectively, were monitored sequentially by flow-cytometry, demonstrating a selective inhibition of GlyA-positive cells following B19-inoculation. Typical morphological changes were observed on cytocentrifuge-spots, and typical giant-cells were identified as staining for GlyA. Productive infection by B19 was demonstrable, as B19-DNA increased by about x 100 after 72 h of culture. The liquid culture method generating erythroid target cells for effective infection by B19 virus promises to be a useful and easily accessible tool for further research on B19 infection of haemopoietic cells.

We characterized the 5' end and parts of the structural genes of European isolates of hepatitis C virus (HCV) and compared them with recently published RNA sequences of American and Japanese HCV isolates. The cDNA, obtained by reverse transcription of viral RNA extracted from different sera, was amplified by nested PCR, cloned and sequenced. Within 239 nucleotides (nt) of the 5' end, we found only three single-nt exchanges compared to two sequences of Japanese origin and one exchange to the prototype HCV sequence (ptHCV) (homology greater than 99%). The sequence of the core region (534 nt) in two European isolates showed a homology of about 97-98% on the nt level, as compared to ptHCV and one Japanese isolate, and 90% to other Japanese isolates. The amino acid (aa) homology was between 98-99% among all published sequences. A greater discrepancy was found in the European isolates within the 434 nt sequenced from the N-terminus of the putative envelope region, where the nt homology to ptHCV and one Japanese isolate was 90-93% (aa homology 93-95%), and to other Japanese isolates was 72-73% (aa homology 77-78%), indicating that the European isolates may be more closely related to the ptHCV and one Japanese isolate than to the other Japanese isolates. Amplified genes encoding structural proteins (core, envelope) were expressed in Escherichia coli. Sera from chronically infected patients reacted strongly with the recombinant core protein, but no specific immunoreactivity occurred with the putative envelope protein.

Six overlapping genomic regions of capsid proteins VP1 and VP3 of hepatitis A virus (HAV) inserted into the expression vectors pBD or pUR respectively expressed beta-galactosidase-HAV fusion proteins. The recombinant proteins were poorly soluble so they were difficult to detect by human anti-HAV sera in radioimmunoassay, but the fusion proteins dissolved in sodium dodecyl sulfate reacted with human and rabbit anti-HAV-positive sera in immunoblots. Antisera against VP1 and VP3 recombinant proteins reacted with the respective structural proteins of HAV in immunoblots. Two recombinant proteins, one including the first 120 amino acids of the N-terminus of VP1 and the other containing all of VP1 except for the first 60 N-terminal amino acids, induced a transient neutralizing antibody response in rabbits. Antisera directed against other regions of VP1 and VP3 neither neutralized viral infectivity nor recognized native virus in a competitive radioimmunoassay. However, when immunized animals were challenged with a sub-immunogenic dose of HAV, all animals responded with stable virus-neutralizing antibodies.

A total of 114 healthy young adults were immunized with hepatitis A vaccine using different vaccination schedules. Individuals received either a single dose (group 1), two doses given simultaneously (group 2), two doses at days 0 and 14 (group 3) or at days 0 and 28 (group 4), or three doses at days 0, 7 and 21 (group 5). Two weeks after a single dose, seroconversion rates between 77 and 85% were achieved (groups 1, 3, 4). All individuals immunized with two doses within two weeks (groups 2, 3, 5) had antibodies to hepatitis A vaccine (anti-HAV positive) by week 3; these participants also showed clearly higher mean anti-HAV values (geometric mean titres, GMTs) at this time than those individuals vaccinated only once. GMTs at week 8 were 560 IU/l in group 5, 236, 339 and 428 IU/l in groups 2–4 and 102 IU/l in group 1. Of participants with anti-HAV at week 8, 82 were again tested 4 months later; all were still seropositive. Ten individuals were tested during the first three weeks at 3–4 day intervals for anti-HAV immunoglobulin M (IgM); specific IgM responses were not detectable before day 10 but were present in eight of 10 vaccinees by day 14.

The extent of human T-cell lymphotropic retorvirus HTLV-I and HTLV-II infections in the general population in central Europe has not been investigated fully. Two hundred forty-eight thousand blood donors from southern Germany were examined serologically for antibodies to the human lymphotropic retroviruses HTLV-I and HTLV-II: 0.021% were confirmed postive and 0.056% were “indeterminate”. A limited number of seropositives and “indeterminate” samples were analyzed by polymerase chain reaction (PCR): the seropositives were confirmed as positive and 43% of the “indeterminate” samples were PCR-positive. The range of 0.021% HTLV-positives in 248,000 donors, i.e. about two in 10,000 individuals, mirrors closely the published data for the United States. © 1994 Wiley-Liss, Inc.

The immunogenicity and reactogenicity of different doses of hepatitis A vaccine was studied in healthy adult volunteers. Vaccinees (105) were immunized with 6.25, 12.5 or 25 ng of HAV antigen, each dose administered at 0, 1 and 6 months (groups B, C and D); one group (group A) obtained three 6.25 ng doses at 0, 1 and 2 months. After one single dose high seroconversion rates ranging between 63 and 85% were observed in all four groups. All participants had seroconverted after the third dose, irrespective of the antigen content per dose and the vaccination schedule. Geometric mean titers after three doses were 439 IU/l (group A, month 3) and 1492, 963 and 2772 IU/l in groups B, C and D at month 7. One year after the first injection all vaccinees tested still showed antibody levels well above 10 IU/l. The vaccine was very well tolerated. Minor localized symptoms were observed mainly such as slight pain at the injection site. These symptoms were not dose related; no serious side effects occurred.

Infectious cellular uptake of human immunodeficiency virus (HIV) is initiated by a complex sequence of interactions between the viral envelope gp120/gp41 complex and the cellular CD4 receptor resulting in the exposure of a hydrophobic region of gp41 that mediates the irreversible fusion of the virus with the cell membrane. Here we show that viral penetration into a susceptible cell can be inhibited by the high-affinity monoclonal CD4 antibody (CD4 mAb) M-T413 even when it is added as late as 30-120 min after the initial contact of virus with the cell membrane. Inhibition of infection was assessed by monitoring cultures for 34 days after exposure to virus using four different methods simultaneously, including detection of viral DNA by PCR. The interval during which HIV remains sensitive to postbinding neutralization by CD4 mAb depends on strain of virus and type of target cell. Preparations of recombinant soluble CD4 (and the immunoadhesin CD4-IgG1) were much less efficient when compared with mAb M-T413, particularly in blocking infection by fresh HIV-1 isolates. Also cellular transmission of HIV, as determined by syncytia formation within 24 hr, was prevented by mAb M-T413 when added within 45 min of contact of infected H9 cells with uninfected C8166 cells. Together with the favorable clinical experience obtained with CD4 mAbs as immunomodulatory drugs, these data suggest that infusion of CD4 mAb M-T413 may be a therapeutic modus for immediate prophylactic intervention after occupational exposure to HIV and for prevention of intrapartum mother-to-infant HIV transmission.

Heat inactivation of parvovirus B19 (B19) was studied in a culture of hematopoietic progenitor cells generated in vitro from peripheral human blood. After inoculating cell cultures with identical volumes of plasma (MII) containing B19 (B19-MII) heat-treated (60 degrees C) for various periods of time, a time-dependent inactivation of the input virus was determined by a decrease of viral DNA replication. No B19 DNA was detected after infection with B19-MII heat-treated for 20 min or more by Southern blot. Viral B19 protein production decreased time-dependently and was not detected after infection with samples treated for 12 min at 60 degrees C or more determined by the enzyme immunoassay. This study indicates that infectivity of B19 virus in plasma can be reduced in vitro by heat-treatment (60 degrees C). However, this does not mean that the heat treatment completely inactivated B19 virus.

Erythroid progenitor cells generated in vitro from peripheral human blood in the presence of interleukin-3 and erythropoietin were infected with human parvovirus B19. B19 virus DNA replication was highest 48 to 72 h after infection, and maximum levels of B19 virus proteins were detected in culture supernatants at 72 to 96 h after infection. B19 virus propagated in vitro was infectious. This cell culture system with peripheral blood cells facilitates studies in vitro of B19 virus replication.

A cDNA fragment corresponding to the nonstructural gene region of Hepatitis C virus was cloned and sequenced. cDNA was obtained by reverse transcription of viral RNA extracted from serum of a German patient with chronic post transfusion hepatitis. “Nested” PCR resulted in a cDNA fragment of 345 nt. The sequence showed a homology of 96% to the American prototype HCV.

A seroepidemiological study on the prevalence of antibodies against heptitis A virus (HAV), hepatitis B virus (HBV) and Treponema pallidum was conducted in various groups of the population of the state of Mauritius (Islands of Mauritius and Rodrigues). 618 sera were tested. The overall prevalence of anti-HAV was 86.1% and yielded an age-dependent increase. Serological evidence for acute or chronic HBV infection was found in 3.8%; 4.5% were positive for anti-HBc alone, and in 12.6% past HBV infection was detected. No age- or sex-dependent increase in the prevalence of anti-HBc was found. There were differences in the anti-HBc prevalence among the various groups of population ranging from 5.9 (flight personnel) to 58.3% (prison inmates). Treponemal antibodies were detected in 6.0% and showed a fairly marked age-dependent increase. Our study suggests that vaccination programmes against HAV and HBV would be beneficial for the Mauritian population.

Infectious herpes simplex virus type 1 (HSV-1) recombinants were constructed by inserting the cDNA sequence of the human immunodeficiency virus type 1 (HIV-1) gag gene (from nucleotide position 675 [SacI] to 3859 [Asp718] of the cDNA sequences of HIV-1 strain BH-10) within the DNA sequences of the BamHI DNA fragment B of the genome of an apathogenic HSV-1 strain HFEM. This HSV-1 strain possesses a 4.1-kbp deletion within the BamHI DNA fragment B between 0.762 and 0.789 map units of the viral genome, which allows the insertion of at least 4 kbp of foreign genetic material into this particular region. The DNA sequences of the immediate early promoter (IE4) of HSV-1 that were inserted upstream from the gag gene were used as a promoter. The screening of 205 virus stocks derived from individual plaques revealed that 46 recombinant viruses harbor HIV-1 gag-specific DNA sequences. However, it was found that only six of the recombinant viruses are able to express the gag gene product of HIV-1. This indicates that the ratio of the positive recombination events is about 2.9%.

A newly developed recombinant antigen-based anti-HIV-1/HIV-2 enzyme immunoassay (Abbott Recombinant HIV-1/HIV-2 EIA) was evaluated against a second generation anti-HIV-1 EIA (Abbott Recombinant HIV-1 EIA). Five thousand and twenty-nine sera from European blood donors and 403 sera from central African blood donors were used in the evaluation, along with four panels and one cohort. The panels included 99 'problem' sera, 733 sera with antibodies to HIV-1 from asymptomatic people and from patients at different disease stages, 25 serial bleeds from five plasmapheresis donors seroconverting for antibodies to HIV-1, and 202 sera with antibodies to HIV-2 collected from healthy and diseased people of European or west African origin. In addition, 734 sera collected from a west African cohort were tested. Using Western blot as the reference standard, the specificity obtained by the recombinant anti-HIV-1 EIA (HIV-i EIA) was 99.90% [99.81-99.99%; 95% confidence limits (95% CL)] with European blood donor sera; 99.50% (98.78-100%) with Central Africa blood donor sera; 92.93% (87.78-98.08%) with 'problem' sera and 99.43% (98.87-100%) with sera from a west African cohort. Using the same samples, the recombinant anti-HIV-1/HIV-2 EIA (HIV-1/HIV-2 EIA) yielded a specificity of 99.84% (99.73-99.95%), 99.50% (98.78-100%), 95.96% (92.00-99.92%) and 98.58% (97.69-99.47%), respectively. All 776 Western blot-confirmed anti-HIV-1 sera were reactive in both EIAs, and the EIA-reactive samples from seroconverting plasma donors were always observed for both assays in the same serial bleed. For HIV-2, the HIV-1 EIA yielded an overall sensitivity of 75.83% (69.93-81.72%) compared with 99.53% (98.58-100%) for HIV-1/HIV-2 EIA. The addition of a recombinant env-protein of HIV-2 to the recombinant env and core proteins of HIV-1 on the solid phase of HIV-1 EIA improved the detection of anti-HIV-2 while preserving the assay's overall specificity and sensitivity for the detection of anti-HIV-1.

The natural course of chronic hepatitis non-A, non-B (HNANB) was documented for 3-20 yr (mean 8 yr) in 86 patients, who attended our special ambulance between 1981 and 1988. Sixty five of the 86 patients (75%) were positive for circulating antibodies against hepatitis C virus (HCV) (anti-HCV). Twenty four patients had chronic posttransfusion (PT)-HNANB (18 anti-HCV-positive; 75%), and 62 patients had sporadic (S)-HNANB (47 anti-HCV-positive; 75%). Twenty nine per cent of patients with chronic PT-HNANB had sustained normalization of aminotransferases after a period up to 5 yr, 55% demonstrated chronic persistent hepatitis (CPH) and 16% progressed to chronic active hepatitis (CAH) with transition to cirrhosis. In the group with chronic S-HNANB, 2% of patients showed remission, 43% had stable CPH and 55% progressed to CAH or cirrhosis. However, development of cirrhotic complications required many years. Transition from CAH to CPH or remission was not observed. The results indicate that 75% of both patients groups with chronic PT- and S-HNANB are infected with the same agent, of which antibodies are detected by the new anti-HCV assay. There was no statistical association between the severity of the disease and the presence of anti-HCV. The different proportions of progressive courses in chronic PT- and S-HNANB might be explained by the patient recruitment.

Hepatitis A vaccine, strain HM175, was investigated for immunogenicity and tolerability in a prospective multicentre trial. The following vaccination schedules and antigen contents were evaluated: days 0 and 14 with 720 ELISA units (El.U) of antigen, days 0 and 28 with 720 El.U and days 0 and 28 with 360 El.U. In all study groups, the seroconversion rates following two vaccinations were between 95 and 100%. Higher geometric mean concentrations of antibody to hepatitis A virus (anti-HAV) were reached by the vaccine containing 720 El.U of HAV antigen. The vaccine was equally well tolerated in all groups. In addition, an abbreviated schedule, in which 720 El.U of HAV antigen was given on days 0 and 14, resulted in 100% seroconversion by day 28 and a level of anti-HAV that was substantially higher than that observed after passive immunization. This implies that such a vaccine could replace immune globulin administration if time permits.

(1990). Transmission of hepatitis C virus (HCV) from a Haemodialysis Patient to a Medical Staff Member. Scandinavian Journal of Infectious Diseases: Vol. 22, No. 6, pp. 757-758.

Two hundred and twelve individuals who failed to respond or responded poorly to hepatitis B vaccination or whose anti-HBs levels had dropped below 10 IU/L at follow-up were revaccinated. Only 18% of initial nonresponders, but 96% of those who responded initially, developed anti-HBs above 10 IU/L after revaccination. In 85% of vaccinees, anti-HBs titres after a fourth immunization were significantly higher than after completion of the first full immunization course, and despite a steeper decline of antibody levels after revaccination, anti-HBs seemed to persist better than after the initial course of three inoculations. All individuals who responded initially and in whom anti-HBs had become undetectable developed antibodies. Response to booster doses was seen after three to five days, and a fifth inoculation given to 23 individuals induced even better responses.

A virus-transformed marmoset lymphoblastoid cell line (LCL) with T-cell characteristics was used for the production of rabbit anti-marmoset T-cell globulin (RAMTG). Exhaustive absorption of the crude antiserum with kidney-liver homogenate and pooled marmoset B-LCL yielded a T-cell-specific antibody. Specificity of the absorbed RAMTG was evidenced by plateau formation in the cytotoxic activity on marmoset peripheral blood lymphocytes (PBL) and clear discrimination between marmoset T-LCL and B-LCL. Unexpectedly, the absorbed RAMTG was not cytotoxic for human PBL. Assays for mitogenic activity showed that unabsorbed RAMTG exhibited mitogenic activity on marmoset, but not on human or baboon PBL. Unabsorbed rabbit anti-human T-cell globulin (RAHTG) induced cellular proliferation in human and baboon PBL, whereas marmoset or Saimiri sciureus PBL showed no response. Absorbed RAMTG did not stimulate DNA synthesis of marmoset PBL. In conclusion, the specific RAMTG may be useful for studies on the functional role of T-lymphocytes in the marmoset immune response to organ allografts or infectious agents.

Using a solid phase radioimmunoassay, antibody to hepatitis A virus (anti-HAV) was determined in 3890 sera from populations in seven European countries. Prevalence of anti-HAV was lowest in Scandinavian countries and highest in Greece and France. Antibodies were found in 77 (13%) of 602 blood donors in Sweden, in 29 (17%) of 175 blood donors and women taking birth control pills in Norway, in 273 (39%) of 700 blood donors in Switzerland, in 262 (52%) of 505 blood donors in Holland, in 365 (55%) of 661 accident patients in West Germany, in 452 (75%) of 600 blood donors in France and in 530 (82%) of 647 persons in Greece. Prevalence of anti-HAV increased with age in all populations tested, indicating nearly total exposure to HAV in persons over 19 years of age in Greece and in persons over 39 years of age in West Germany, Holland and France. Antibody was found more frequently in rural than in urban populations in Greece and Switzerland. Calculation of the age-specific incidence of HAV infections suggests a remarkable decline in the exposure rate in the last few decades.

human diploid fibroblasts and human primary liver cell carcinoma cells (PLC/PRF/5) were infected with hepatitis A virus (HAV) adapted to growth in cell culture or derived directly from human stool. Viral antigen was expressed in PLC/PRF/5 cells 28 days after infection with cell culture-adapted HAV, and 50 days after infection with virus from human stool. In human fibroblasts the periods until first expression of viral antigen were 90 and 210 days, respectively. During further passages of HAV in fibroblasts the time of first appearance of antigen decreased to about 28 days. Biophysical properties of HAV extracted from infected fibroblasts were comparable to those of HAV derived directly from human stool. Immunofluorescence studies showed that the antigen was located exclusively within the cytoplasm of the infected fibroblasts. Kinetics of antigen production indicated that an equilibrium between virus multiplication and cell metabolism was reached in persistently infected fibroblasts.

Continuous lymphoblastoid cell cultures were established from marmoset (Saguinus sp.), squirrel (Saimiri sciureus), owl (Aotus trivirgatus) and cebus (Cebus apella) monkeys after culturing their peripheral lymphocytes with lethally X-irradiated cells carrying Epstein-Barr virus (EBV). Transformation also was achieved by exposing simian lymphocytes to infectious, cell-free EBV derived from the simian lymphoblastoid cell cultures. Simian lymphocytes were not transformed after exposure to cell-free EBV derived from HR-1 cells. The simian cell cultures were similar to cell cultures derived from Burkitt's lymphoma or infectious mononucleosis patients. EBV-induced early, viral capsid and membrane antigens, intranuclear inclusion bodies and herpesvirus virions were demonstrable in most cultures. Seven cultures were insusceptible to superinfection with EBV and treatment of the cultures with halogenated pyrimidines was relatively ineffective for inducing synthesis of early or viral capsid antigens. All cell cultures had B-cell characteristics: they produced immunoglobulins but did not form spontaneous rosettes with sheep erythrocytes. Four of six marmoset monkeys, inoculated with EBV-transformed marmoset lymphocytes, developed antibodies to EB viral capsid antigens and one marmoset inoculated with autochthonous transformed cells also developed heterophile antibodies. Seven marmosets, inoculated with cell-free EBV derived from HR-1 cell cultures, developed no detectable levels of antibodies to EBV-specified antigens or heterophile antibodies. No overt clinical abnormalities were detected in any of the marmosets inoculated with HR-1 or Kaplan EBV but one of five marmosets inoculated with B95hyphen;8 EBV developed a lymphoma. Virus D'Epstein-Barr: Transformation de Lymphocytes de Primates Non Humains In Vitro Des cultures continues de cellules lymphoblastoides ont ete etablies a partir de ouistitis (Saguinus), de saimiris (Saimiri sciureus), de singes de nuit (Aotus trivirgatus) et de sajous (Cebus apella) apres culture de leurs lymphocytes peripheriques avec des cellules letalement irradiees porteuses du virus d'Epstein-Barr (EBV). La transformation a egalement ete obtenue en exposant des lymphocytes simiens a de l'EBV acellulaire infectieux provenant des cultures de cellules lymphoblastoides simiennes. Les lymphocytes simiens n'etaient pas transformes apres exposition a l'EBV acellulaire provenant des cellules HR-1. Les cultures de cellules simiennes etaient analogues aux cultures provenant de cas de lymphome de Burkitt ou de mononucleose infectieuse. On a observe dans la plupart des cultures des antigenes de la membrane ou des capsides virales et des antigenes precoces induits par l'EBV des inclusions intranucleaires et des virions du virus herpetique. Sept cultures n'etaient pas sensibles a la surinfection par l'EBV et le traitement aux pyrimidines halogenees a ete relativement inefficace du point de vue de l'induction de la synthese des antigenes precoces ou des antigenes des capsides virales. Toutes les cultures avaient les caracteristiques des cellules B: elles produisaient des immunoglobulines mais ne formaient pas spontanement de rosettes avec les erythrocytes de mouton. Quatre des six ouistitis inocules avec des lymphocytes de ouistiti transformes par l'EBV ont developpe des anticorps contre les antigenes des capsides virales EB et un ouistiti inocule avec des cellules autochtones transformees a produit egalement des anticorps heterophiles. Chez sept ouistitis inocules avec de l'EBV acellulaire provenant de cultures de cellules HR-1. on n'a pas observe d'anticorps contre les antigenes determines par l'EBV ou d'anticorps heterophiles. Aucune anomalie clinique manifeste n'a ete decelee chez les ouistitis inocules avec l'EBV des HR-1 ou des cellules de Kaplan, mais l'un des cinq ouistitis inocules avec l'EBV de la culture B95-8 a developpe un lymphome.

A microtechnique for marmoset lymphocyte cultures is described that allows a 10-fold reduction of cell numbers and the amount of culture medium and radioisotopes. Optimal culture conditions for the response of Saguinus fuscicollis, Saguinus oedipus , and Callithrix jacchus peripheral blood lymphocytes to a variety of mitogens were established. The usefulness of marmoset lymphocyte microcultures for the evaluation of alterations of immunocompetence associated with experimental viral disease and its possible application for immunogenetic studies are discussed.

Hepatitis B e antigen (HBeAg) and its antibody (anti-HBe) were evaluated using a sensitive radioimmunoassay (RIA) in weekly serum samples obtained from nine chimpanzees experimentally infected with hepatitis B virus (HBV). In two chimpanzees with HBV infection with detectable hepatitis B surface antigen (HBsAG) for less than five weeks, and in one chimpanzee with documented HBV infection with no detectable HBsAg, HBeAg was not detected; in all three, anti-HBe became detectable early in the infection. In six chimpanzees in which HGsAg was detected for 16 weeks or longer, HBeAg was detected early in the infection; in five, anti-HBe became detectable and HBeAg undetectable prior to the clearance of HBsAg. The sixth remained HGsAg-positive and HBeAg-positive for more than two years and never developed anti-HBe. These results confirmed the sensitivity of this RIA and its value in predicting the course of HBV infections.

The relationship between NK active and IFN alpha-producing cells in human peripheral blood was studied with a monoclonal antibody with specificity for NK cells (anti-Leu11b). Removal of Leu11b antigen expressing leukocytes with antibody-mediated complemeritdependent lysis resulted in a marked reduction of NK activity. In contrast, the depletion of Leu11b positive cells did not affect the production of IFN alpha in response to influenza A/X31 virus, Corynebacterium parvum, or Molt 4 human leukemic cells. The results indicate that NK activity and synthesis of IFN alpha are mediated by different leukocyte subpopulations. The findings further suggest that the augmentation of the cytotoxic activity of NK cells by IFN may not be the consequence of positive self-regulation, but rather of cellular cooperation.

Two panels consisting of 236 and 258 anti-LAV/HTLV III positive and 64 and 50 negative sera, respectively (determined in Western blots), were tested with 8 different, commercially available ELISA test kits for detecting LAV/HTLV III antibodies (Abbott, Dupont, Electro-Nucleonics, Litton, Organon, Pasteur, Sorin, Wellcome). Positive sera were selected for levels of antibodies ranging from average to very high, and the negative sera included both negative sera and sera with antibodies to cellular components such as HLA antigens or nuclear membranes. In examination of the 2 panels, the sensitivity of the tests ranged from 97.2 to 100%, and the specificity from 70.0 to 100%. No test was ideal.

An ELISA for the detection of specific IgM and IgG against human parvovirus B19 (anti-B19 IgM and IgG) and B19 antigen is described. With ELISA anti-B19 IgM could be detected for up to 20 weeks after viraemia. Four to five months after B19 infection anti-B19 IgG titres range between 10−6 and 10−7. Nonspecific reactions with rheumatoid factor or IgM against rubella were not found. The ELISA for B19 antigen was shown to be as sensitive as DNA hybridisation. With immunoblotting two viral proteins of 83 kd (VP1) and 58 kd (VP2) were demonstrated. After acute infection antibodies to VP2 appear before antibodies to VP1. Immunoblotting might be used in pregnancy to determine the time of maternal infection. In a survey of a B19 outbreak in a school for medical technology, 6 (28.6%) of 21 non-immune females seroconverted.

The effects of specific immunotherapy with allogeneic cells transformed by Schmidt-Ruppin Rous sarcoma virus (SR-RSV), of treatment with BCG, and of surgery on the growth of SR-RSV-induced sarcomas in white-lipped marmosets were studied. Tumor incidence, tumor progression, and survival did not differ between control and treated animals. Animals immunized with BCG developed lymphocyte reactivity to tuberculin, which remained until the animals died. BCG was isolated from the spleen of one tumor-bearing animal.

This paper describes the construction of a new heteromyeloma cell line designated CB-F7. The cell line was derived from xenogeneic somatic cell hybridization between normal human B lymphocytes and the murine HAT-sensitive P3X63Ag8/653 cell line. CB-F7 cells were characterized by rapid cell growth (doubling time about 16 h) and high cloning efficiencies in culture medium supplemented with 10% or 5% fetal calf serum, respectively. The karyotype of the cells consists of about 75-78 chromosomes as well as two chromosomal fragments. Fusions of the cells with human peripheral blood cells resulted in approximately 2-6 clones per 10(5) seeded lymphocytes. Furthermore, the cells are ouabain resistant and therefore suitable for fusions with EBV-transformed lymphoblastoid cell lines. Using CB-F7 as the parental cell line a number of specific human mAb producing hybrids were established. For the first time, we describe here the generation of hybrids secreting human monoclonal antibodies to human immunodeficiency virus (HIV). Two monoclonal antibodies of IgG type and one of IgM type reacted with the major core protein p25 and one IgG antibody reacted with the transmembrane protein gp41.

The prevalence of hepatitis δ virus (HDV) was evaluated in 1027 HBsAg positive people living in five provinces, two municipalities and four autonomous regions of China. Seventeen people had antibodies to HDV (anti-HD — 13 of 454 asymptomatic carriers of HBsAg, two of 151 patients with chronic persistent hepatitis and two of 59 patients with chronic active hepatitis). All of those who were anti-HD lived in the autonomous regions of Inner Mongolia, Tibet, and Xinjiang.