Anti-HIV IgM antibody analysis during early manifestations of HIV infections.

In a prospective study we tested the appearance of IgG and IgM positive viral protein bands in Western blots from six people who seroconverted for anti-HIV antibody. Quantification of the immunoblotted bands was performed by reading the Western blot stripes in Camag scanner and analysed on a 350 computer (digital equipment). In the first serum, all people were negative for anti-HIV antibodies. In the second serum, after 16 to 122 days, all people showed IgM HIV-antibodies to p24. IgG HIV-antibodies were detectable in all people after 18 to 114 days after the second collection. Our data clearly demonstrated that for early analysis of HIV infection only the detection of IgM antibodies to viral protein bands of the Western blot technique provides reliable results and that scanning and advanced integration analysis of the Western blot peaks offer the advantage of direct quantitative comparison of the results, not just qualitative description. Further, this direct quantitative comparison of antibodies to HIV virus protein bands can be used as a prognostic marker for disease states.

Highly active antiretroviral therapy during early HIV infection reverses T-cell activation and maturation abnormalities. Swiss HIV Cohort Study.

OBJECTIVES: To evaluate the impact of early initiation of highly active antiretroviral therapy (HAART) on disease-induced T-cell activation and maturation abnormalities during asymptomatic HIV infection. DESIGN: A prospective open-label trial of zidovudine, lamivudine and ritonavir in treatment-naive asymptomatic HIV-infected individuals with CD4 cells > or = 400 x 10(6)/l. METHODS: Peripheral blood CD4+ and CD8+ T cells derived from 15 asymptomatic HIV-infected individuals (median baseline CD4+ cells, 608 x 10(6)/l; CD8+ cells, 894 x 10(6)/l; plasma HIV RNA, 3.93 log10 copies/ml) undergoing therapy with zidovudine (300 mg twice daily), lamivudine (150 mg twice daily), and ritonavir (600 mg twice daily) were assessed for changes in expression of phenotypic markers of T-cell activation (HLA-DR and CD38) and maturation (CD45RA and CD45RO). At weeks 0, 2, 4, 8, 12, 16, 20 and 24, T-cell subsets were quantified by flow cytometry and plasma HIV viral loads determined using reverse transcription PCR. RESULTS: HAART-induced decrease in plasma HIV RNA levels coincided with a significant reduction in numbers of activated CD4+/HLA-DR+ (maximum change, -36%; P < or = 0.05), CD8+/HLA-DR+ (maximum change, -66%; P < or = 0.005) and CD8+/CD38+ (maximum change, -51%; P < or = 0.01) T cells. A concomitant significant increase in numbers of naive CD4+/CD45RA+ (maximum change, +12%; P < or = 0.005) and memory CD4+/CD45RO+ (maximum change, +6%; P < or = 0.05) T cells was also evident, which contrasted with a significant decrease in memory CD8+/CD45RO+ cells (maximum change, -42%; P < or = 0.005). CONCLUSION: The observed ability of HAART during early asymptomatic HIV infection to initiate rapid reversal of disease-induced T-cell activation and maturation abnormalities, while preserving pretherapy levels of immune function, supports the concept that therapeutic advantage is to be gained by commencing early aggressive antiretroviral therapy.

Change in circulating levels of the chemokines macrophage inflammatory proteins 1 alpha and 11 beta, RANTES, monocyte chemotactic protein-1 and interleukin-16 following treatment of severely immunodeficient HIV-infected individuals with indinavir.

OBJECTIVE: To evaluate the in vivo relationship between HIV replication and circulating levels of the chemokines macrophage inflammatory protein (MIP)-1 alpha, MIP-1 beta, RANTES (acronym for Regulated upon Activation, Normal T-cell Expressed and presumably Secreted), interleukin (IL)-16 and monocyte chemotactic protein (MCP)-1, which have recently been characterized as factors capable of regulating in vitro HIV replication. DESIGN AND METHODS: We have compared changes in plasma HIV-RNA levels and circulating levels of MIP-1 alpha, MIP-1 beta, RANTES, IL-16 and MCP-1 in 20 severely immunodeficient HIV-infected individuals (CD4+ T cells = 14 +/- 3 cells x 10(6)/l; plasma HIV RNA = 4.45 +/- 0.27 log 10 copies/ml) undergoing treatment with the HIV protease inhibitor indinavir that added to pre-existing nucleoside-based therapy. At weeks 0, 2, 6 and 12, viral load was quantified using a commercial reverse-transcription polymerase chain reaction assay, peripheral blood T-cell subpopulations assessed by flow cytometry, and chemokine levels quantified using commercial sandwich enzyme-linked immunosorbent assay kits. RESULTS: Following initiation of indinavir-based therapy, significant decreases in plasma HIV-RNA levels (change = 2.0 +/- 0.75 log 10 copies/ml) were observed in conjunction with significant increases in absolute CD4+ (change = 83 +/- 19 cells x 10(6)/l) and CD8+ (change = 293 +/- 96 cells x 10(6)/l) T-cell numbers. Concomitantly, significant increases in MIP-1 alpha (19% increase), MIP-1 beta (14% increase), RANTES (15% increase) and IL-16 (1213% increase) levels occurred. In contrast, MCP-1 levels decreased significantly (47% decrease). CONCLUSION: The in vivo demonstration of an association between diminishing plasma HIV-RNA levels and the emergence of a circulating chemokine profile capable of inhibiting HIV replication corroborates recent in vitro observations and provides evidence for the restoration of chemokine capacity by HIV protease inhibitor-based therapy.

Simple monitoring of antiretroviral therapy with a signal-amplification-boosted HIV-1 p24 antigen assay with heat-denatured plasma.

OBJECTIVE: Virus load determination has become indispensable for the management of HIV patients, but depends on expensive assays of a low throughput. We evaluated whether a highly improved HIV-1 p24 antigen detection procedure which involves heat-mediated immune complex dissociation and signal-amplification-boosted enzyme-linked immunosorbent assay (ELISA) was suitable for antiretroviral treatment monitoring. DESIGN AND METHODS: Virus load in plasma was determined for 127 plasma samples taken at 0, 2, 6, 12, 18, 24, 30 and 36 weeks from 23 patients with CD4+ T cells < 50 x 10(6)/l who received indinavir 800 mg three times daily in addition to prior antiretroviral treatment. Tests included polymerase chain reaction (PCR) for viral RNA, measured prospectively with the Roche Amplicor kit, and retrospective batch testing of heat-denatured samples for p24 antigen by the DuPont HIV-1 p24 Core Profile ELISA linked with a tyramide signal amplification step. Particle-associated reverse transcriptase (RT) by the product-enhanced RT (PERT) assay was determined as an independent third-opinion viral load marker. RESULTS: p24 antigen was detected as sensitively as viral RNA. Overall detection during a median observation time of 25 weeks (range, 0-39) amounted to 75.6% for antigen and 73.6% for RNA. The antigen detection limit was 0.2 pg/ml. Antigen was detectable in all 23 baseline samples, whereas RNA was undetectable in one. Antigen and RNA levels in 79 samples positive for both markers correlated with r = 0.714 (P < 0.0001). Average changes in levels of p24 antigen and RNA at eight timepoints correlated with r = 0.982 (P < 0.0001). In individual patients, the two parameters behaved similarly, and in certain cases virtually identically. RT activity was measurable in all samples. CONCLUSIONS: The performance of this antigen detection procedure is comparable to RNA PCR, thus providing a simple, high throughput alternative in monitoring the efficacy of antiretroviral treatment.

Recombinant gp160 as a therapeutic vaccine for HIV-infection: results of a large randomized, controlled trial. European Multinational IMMUNO AIDS Vaccine Study Group.

OBJECTIVES: The primary objective of this study was to expand the safety and immunogenicity database of recombinant gp160 as a therapeutic vaccine in the treatment of HIV-infection. Preliminary efficacy data was also sought. DESIGN: This trial was a randomized, double-blind, placebo-controlled study. Two-hundred and eight volunteers, 96 therapy-naive with CD4 cell count >500x10(6)/l (group A) and 112 with CD4 cell count of 200-500x10(6)/l (group B, 51 out of 112 on treatment with one or two nucleoside analogues), received monthly injections of rgp160 IIIB vaccine or placebo for the first 6 months of the study; booster immunizations with rgp160 MN or placebo were given at times 15, 18, and 21 months. METHODS: Safety and immunogenicity data were obtained and measurements of CD4 cell count, plasma viral RNA, and proviral DNA were performed. Clinical outcome was recorded for the 24 months of study. RESULTS: The vaccine was safe and well tolerated. Despite the induction of new rgp160-specific lymphoproliferative responses and the presence of positive delayed type hypersensitivity skin tests to rgp160 at the end of the 24 month study, no effect on the natural history of HIV infection was detected. Within 24 months, AIDS-defining illnesses had occurred in 19 of the vaccinated volunteers and in 18 of the placebo recipients. Persons with higher plasma viral RNA levels and higher proviral DNA had a more rapid decline in CD4 cell count when compared to persons with lower values. Vaccine did not alter viral RNA or proviral DNA levels. CONCLUSION: There was no clinical benefit to therapeutic immunizations with rgp160, despite the induction of new lymphoproliferative responses.

Glia cell stimulating factor (GSF): a new lymphokine. Part 1. Cellular sources and partial purification of murine GSF, role of cytoskeleton and protein synthesis in its production.

The effect of activated-lymphocyte supernatant on glia cells was investigated. When treated in vitro with Concanavalin A (ConA), murine spleen cells released a soluble product, termed glia cell stimulating factor (GSF), which stimulated RNA and DNA synthesis in cultured murine glia cells. Furthermore, GSF appeared to promote the maturation of undifferentiated glia cells to astrocytes having a high content of glial fibrillary acidic protein. GSF secretion occurred after a lag period of 16 hours and proceeded at a constant rate for more than 48 hours. This GSF produced by ConA-stimulated murine lymphocytes has an apparent molecular weight between 60,000 and 80,000. Antigenic stimulation of primed lymph node cells with BGG resulted in a similar GSF production. Cellular sources of mitogen-induced GSF were investigated by using isolated lymphoid populations. GSF release by ConA-activated pure T-lymphocytes reconstituted with peritoneal macrophages was equivalent to that of unseparated spleen cells, whereas GSF production by T-lymphocytes alone was low. Macrophages alone did not elaborate detectable levels of GSF. GSF was also secreted by enriched -B-lymphocytes populations stimulated by Protein A. Formation of GSF was suppressed when cytochalasin B or cyclo-heximide was added to the cultures, while colchicine failed to have any effect. DNA synthesis is not required for GSF production as determined by resistence to treatment with mitomycin C. The data indicate that the GSF production and secretion mechanism is much like that described for other lymphokines.

Glia cell stimulating factor (GSF): a new lymphokine. Part 2. Cellular sources and partial purification of human GSF.

Glia cell proliferation is characteristic of many inflammatory and degenerative processes in the brain, however the mechanisms underlying this response are poorly understood. We described a glia cell stimulating factor (GSF), produced by murine spleen cells, which activates DNA- and RNA synthesis of cultured glia cells. In the present series of experiments, we examined the ability of Concanavalin A (ConA)-stimulated human peripheral blood mononuclear leukocytes (PBM) and two continuous cell lines derived from human lymphocytes to spontaneously release of GSF in culture. The studies presented herein demonstrate that (1) ConA-stimulated PBM of healthy subjects produced GSF, (2) GSF is produced by T lymphocytes in collaboration with monocyte-macrophages, (3) both the human T cell line (MOLT-4F) and a B cell line (RPMI 1788) spontaneously secrete GSF, (4) GSF was found to have a molecular weight of 30,000 and less than 10,000 daltons, (5) macrophage (MIF)- and leukocyte (LIF) migration-inhibiting factor activities, as well as mitogenic factor (MF)- and lymphocyte-activation factor (LAF) activities could be separated from the GSF activity by gel filtration on Biogel P-100. These findings provide further evidence for the existence of GSF as a new lymphokine, distinct from LIF, MIF, MF and LAF.

Dual effect of glia maturation factor on astrocytes. Differentiation and release of interleukin-1 like factors.

C6 glioma cells, and primary cultures of mouse astrocytes, stimulated with lipopolysaccharide (LPS) release an interleukin-1 like factor (IL-1) which enhances lectin-induced T-lymphocyte proliferation and promotes the release of interleukin-2 (IL-2) by ConA-stimulated thymocytes. In the present study, the glia maturation factor (GMF) was found not only to induce differentiation of glioblasts, but also to elicit the secretion of IL-1 like factors by cultured mouse astrocytes and their precursor cells. GMF was also effective in triggering IL-1 release by macrophages. Contamination of the 23 000 MW GMF preparation with LPS was excluded by the Limulus lysate assay and by using C3H/HeJ LPS-nonresponder mice whose glia and macrophages responded to GMF but not to LPS, by IL-1 release. Through its ability to induce glial differentiation and IL-1 release, GMF may represent an important endogenous signal, triggering both reactive gliosis and the development of an immune response within the central nervous system.

Immunogenicity of a hepatitis B subunit vaccine in hemodialysis and in renal transplant recipients.

A hepatitis B subunit vaccine was given to 59 medical staff members, 106 hemodialysis patients and 28 renal allograft recipients. The vaccine consisted of formalin-inactivated hepatitis Bsurface antigen (HBsAg) and was given in 3 doses (times 0, 1 and 6 months) of 20-40 micrograms. Some of the vaccinees received anti-HBs antibodies together with the first vaccine dose (active/passive vaccination). One month after the last infection, 93% of the medical staff members who had received active/passive immunisation and 97% of those who had received active immunisation had detectable anti-HBs antibodies with mean titers ranging from 1:512 to 1:1024. In the group of hemodialysis patients antibodies were detectable in 63-65% of the individuals who had received active or passive/active immunisation in mean titers between 1:32 and 1:64. Finally, only 32% of the renal allograft patients developed measurable anti-HBs antibodies, the titers of responders being still lower than in the hemodialysis patients. Side effects occurred following 10% of all vaccine injections and were always mild in nature. Within the 12 months observation period period following the first vaccination, 3 HBV events occurred in the 193 individuals: One aclinical case detected by a transient seroconversion against the hepatitis B core antigen, one anicteric and one icteric hepatitis case. The data illustrate the difficulties for active immunisation against hepatitis B of hemodialysis patients or of renal transplant recipients.

Chronic viral hepatitis B and C: an argument against the conventional classification of chronic hepatitis.

The classification of chronic hepatitis distinguishing benign chronic persistent hepatitis from severe chronic active hepatitis was constructed without knowledge of well-defined aetiological factors. Better understanding of the different hepatitis-viruses has shed new light on this subject. Chronic viral hepatitis B and C each show typical histological patterns. The validity of the conventional classification has been evaluated by a comparative study of chronic viral hepatitis B and C. 130 biopsies from 110 patients with chronic hepatitis C (CH-C) proven serologically by antibodies (second generation testing) were compared with 105 biopsies from 73 patients with chronic hepatitis B (CH-B). These were scored semi-quantatively. In CH-C, lymphoid follicles and/or aggregates were found in 88.5%, fatty degeneration in 51%, bile duct lesions in 46.2%, and Mallory body-like material in the hepatocytes in 9.2%. The portal lymphocytic infiltration generally predominated over the necro-inflammatory lesions of the parenchyma. Chronic persistent hepatitis (defined by the presence of portal hepatitis) was present exclusively in CH-C. Chronic lobular hepatitis was found exclusively in CH-B. We conclude that the histological criteria described for CH-C are highly suggestive of the diagnosis, that the artificial subdivision of chronic hepatitis into CPH and CAH is obsolete and that the histological assessment of chronic hepatitis should consist of a grading of inflammatory activity (minimal, mild, moderate, severe) and staging of fibrosis (extent of distortion of architecture). The final diagnosis should be based on the demonstration of the aetiological agent.

Polyvalent immunoglobulins for prophylaxis of bacterial infections in patients following multiple trauma. A randomized, placebo-controlled study.

One hundred and fifty severely injured patients requiring long-term artificial ventilation were evaluated in a prospective, randomized, double blind study comparing the prophylactic effect of an intravenous immunoglobulin (Sandoglobulin; IGIV) against nosocomial infections with a placebo preparation. The groups were comparable in age, sex, injury pattern, and severity of the trauma. Seventy-six patients received 12 g of Sandoglobulin as a 3% solution on day 0, day 5 and day 12, i.e. a total of 36 g. Sandoglobulin significantly reduced the incidence of pneumonia (28 cases in the IGIV group, 43 cases in the placebo group, p = 0.0111). This resulted in a reduced therapeutic use of antibiotics. For the occurrence of sepsis (IGIV: 14 cases; placebo 19 cases) and other infections (IGIV: 11 cases; placebo: 10 cases) no significant differences were found. No side effects of the administration of IGIV were observed. IGIV prophylaxis neither reduced the overall death rate nor those deaths caused by infection. On day 5 after administration of the first 12 g of IGIV, the IgG serum concentrations were significantly higher in the Sandoglobulin group (8.41 +/- 1.96 mg/ml and 7.42 +/- 2.25 mg/ml respectively, p less than 0.001) whereas later serum samples showed no significant differences.

Immunodeficiency in epilepsy: a new view.

Up to 12% of epileptic patients have subnormal IgA serum concentrations. Previous observations suggest that IgA deficiency is correlated with hydantoin treatment and also with the type of seizure. In a followup study it is shown that IgA deficiency in epileptics is a rather constant feature of a given patient. The most pronounced changes in IgA levels were seen in patients in whom the hydantoin medication also changed. However, low IgA levels have also been reported in untreated epileptics. A new classification for the immunodeficiency state in epileptics is introduced.

Immunoglobulin abnormalities in relatives of IgA deficient epileptics.

Serum levels of IgA were found to be reduced in some patients with epilepsy. Further studies revealed that only epileptics with constitutional factors for seizures showed, if ever, IgA deficiency, particularly those treated with hydantoins (up to 25%). In order further to substantiate the association of immunoglobulin alterations with epilepsy nine families in whom the disease was clustered were investigated. An IgA deficiency was detected in 16 of the 19 epileptics (three without hydantoin medication), but in none of their 45 non-epileptic relatives. However, four of the relatives had a low IgM. Seven other families were tested in each of which only one IgA deficient epileptic was known. No other family members were found with a low IgA, but 24 of 58 such relatives had increased IgM serum concentrations. The association of IgA deficiency and epilepsy with IgM imbalances in relatives of IgA deficient epileptics gives additional support for the hypothesis that immune imbalances and certain forms of epilepsy might be linked.

Transfer factor therapy in patients with subacute sclerosing panencephalitis.

Thirteen patients with subacute sclerosing panencephalitis (S.S.P.E.) at different stages of the disease were admitted for transfer factor treatment. The transfer factor was prepared from non-selected blood bank donors. The activity of the transfer factor was tested in patients with diseases other than S.S.P.E. and was found to be either clinically or immunologically active. Regardless of the number of transfer factor units applied a significant influence on the course of the disease was not apparant. The observed intermittant improvement of 3 patients was considered as spontaneous remission which is known to occur occasionally in S.S.P.E. The humoral and cellular immune response before and after transfer factor therapy did not reveal significant changes which could be correlated with transfer factor therapy.

[Alpha-1-fetoprotein screening--development and problems]. / Alpha-1-Fetoprotein-Screening--Ablauf und Fragestellungen.

Principle and logistic of alpha-1-fetoprotein (AFP-) screening is explained. It is estimated that approximately 2% of women pregnant for 16 to 18 weeks have elevated circulating AFP. By again measuring serum-AFP and by ultrasound examination such abnormal values can be explained by multiple pregnancies, wrongly estimated duration of pregnancy, abortus, etc, in approximately half of the cases. In 0.8 to 1% of all pregnant women the high-serum-AFP remains unexplained and an amniocentesis is indicated. An elevated amniotic AFP (0.1 to 0.2% of all pregnant women) strongly points to a fetus with an open neural tube defect. By also measuring amniotic acetylcholinesterase, this diagnosis can be distinguished from other fetal abnormalities associated with high amniotic AFP, such as omphalocele.

[Hepatitis B virus and pathogenesis]. / Hepatitis-B-Virus und Pathogenese.

The hepatitis B virus belongs to the hepadna viruses family. Its genome consists of an incompletely double stranded DNA. The preS/S domain encodes proteins which make up the outer viral coat containing the HBs surface antigen (HBsAg). Other viral genes programme for structures inside the virus and for various regulatory enzymes. HBV mainly infects hepatocytes. The virus replicates in the cytoplasm and is primarily non-cytopathogenic. HBV can also integrate into the host cell. Various stable genotypes and subtypes are known, which have a characteristic geographic distribution. They all share a common HBsAg epitop, which has allowed the development of a vaccine which is efficient world-wide. The protective principle consists of inducing protective anti-HBs. The infected cell has to be destroyed to eliminate the virus. Cellular immune defence mechanisms are mainly relevant, the principle effectors being cytotoxic T lymphocytes, activated monocytes/macrophages and cytokines such as interferon-gamma. The natural course of infection is highly variable, comprising viral elimination with or without acute hepatitis and chronic infection which might lead to chronic hepatitis, liver cirrhosis and hepatocellular carcinoma. This is due to the balance respectively to the inbalance between the viral replication capacity and the immune defence mechanisms.

[Serologic diagnosis and follow-up of hepatitis B]. / Serologische Diagnose und Verlaufsbeurteilung der Hepatitis B.

This paper is a short summary on the usefulness of two antigens (HBsAg and HBeAg), three antibodies (anti-HBc, anti-HBe and anti-HBs) and of HBV DNA, as markers for the diagnosis and the follow-up of hepatitis B. The significance of each of these markers at the various stages of disease history, a few patterns of co-existence of some of these markers and the occurrence of mutations in the core and pre-core regions of the genome are also described. The various indications for measuring HBV DNA, in addition to the classical serological markers, are also mentioned.

[Street vaccination]. / Gassenimpfung.

Since 1982 drug addicts in the Canton of Zurich have been vaccinated against hepatitis B (HB) free of charge. Up to December 1983, 1480 doses of vaccine were administered to 1270 individuals (an average of 1.2 doses per person). About 700 vaccinations took place directly on the streets of Zurich. The article will discuss the epidemiological data of HB in the drug addict population and also the methodology and difficulties of the Streetwork Vaccination. In addition to immunization, the most important requisite for lowering the incidence of HB would be the possibility for drug addicts to buy injection material themselves.

Hepatitis virus infection in flying airline personnel.

Sera of 1126 flying personnel of an airline were tested for signs of ongoing or past infections with hepatitis B virus (HBV) or with hepatitis A virus (HAV). The prevalence of anti-HA antibodies was similar in all professional categories of flying personnel and the same or slightly lower than in Swiss blood donors. The frequency of immune markers identifying HBV immunity was similar in pilots, flight-engineers, and female flight attendants compared to Swiss blood donors. However, HBV immunity was clearly more prevalent in male flight attendants. Within 1 year, 13 of 2624 flying personnel had acute hepatitis. This higher-than-average incidence of hepatitis amongst flying personnel compared to the Swiss population was mainly due to a high incidence of hepatitis B amongst male flight attendants. Their special life-styles might be responsible for the high prevalence of HBV immunity and for the high incidence of hepatitis B.

[Hepatitis serology: use and interpretation]. / Hepatitisserologie: Anwendung und Interpretation.

Presently five viruses causing hepatitis are known, the hepatitis viruses A (HAV), B (HBV), C (HCV), D (HDV) and E (HEV). The genomic structure is known of most of all these viruses as well as some of their structural and regulatory gene products. Using radio- and enzyme immunoassays viral antigens can be detected for HBV and HDV as well as specific antibodies against all the five viruses. The results of these tests are the basis for the diagnosis and the follow-up of these infections but differ in their accuracy for each given virus. Concerning HIV one can differentiate between an ongoing or recently passed infection and immunity. Concerning HBV (and HDV), an ongoing infection at various stages can be distinguished from immunity. Such distinctions are not possible with respect to HCV except when also applying the expensive and cumbersome method of the polymerase chain reaction. In this paper the most important characteristics of the hepatitis viruses are given and the behaviour of the various viral markers during the infections and their consequences are described.