HTLV-III antibodies in hemodialysis patients — A consequence of blood transfusions?

Summary An investigation for HTLV-III antibodies in chronic hemodialysis patients revealed in four out of 276 patients a positive result using the ELISA and western blot techniques. All HTLV-III positive patients had received blood transfusions. As it has been shown that a needle stick could transmit the HTLV-III, it is suggested that hemodialysis patients who have received frequent blood transfusions should be screened.Abbreviations AIDS acquired immune deficiency syndrome - ELISA enzyme linked immunosorbent assay - HbSAg hepatitis surface antigen - HD hemodialysis - HTLV-III human T-cell lymphotropic virus type III     

Seroepidemiology of HTLV-III (LAV) in the Federal Republic of Germany

Summary In 1984 10,281 sera were collected in the FRG and examined for antibodies to HTLV-III (LAV) with an enzyme-linked immunosorbent assay and confirmative tests. Of the German AIDS patients 81% have antibodies. Individuals belonging to AIDS risk groups, homosexuals, haemophiliacs and i.v. drug abusers, have antibody frequencies between 25%–72%. The detection of HTLV-III antibodies in blood donours indicates that the virus is being transmitted by blood transfusions.Abbreviations AIDS acquired immunodeficiency syndrome - LAS lymphadenopathy syndrome - ARC AIDS related complex - LAV lymphadenopathy associated virus - HTLV-III human T-lymphotropic virus type III - HBV hepatitis B virus     

Counterimmunoelectrophoresis for detection of human serum antibody to HTLV-III

We examined the usefulness of a counterimmunoelectrophoresis (CIE) technique for detecting antibodies to HTLV-III using sera that previously had been assessed for antibodies to HTLV-III by the standard enzyme-linked immunosorbent assay (ELISA). We selected a subset of 53 sera from patients with the acquired immune deficiency syndrome (AIDS) or the generalized lymphadenopathy syndrome (GLS) in which 81.1% were initially ELISA-positive, and 96.2% were positive by Western blot technique. In our standard HTLV-III CIE technique, 58.5% were positive and repeat testing increased the yield to 67.9%. Varying several parameters of the standard CIE assay did not improve sensitivity. We also studied 20 ELISA-negative and 10 ELISA-borderline sera from normal controls; all were negative by CIE. These results indicate that CIE may be used for detection of human serum antibodies to HTLV-III, but that the present assay was less sensitive than the HTLV-III ELISA.     

Intra-blood-brain-barrier synthesis of HTLV-III-specific IgG in patients with neurologic symptoms associated with AIDS or AIDS-related complex

Intra-blood-brain-barrier production of virus-specific antibody is good evidence of infection within the blood-brain barrier. Patients with the acquired immuno-deficiency syndrome (AIDS) have an increased incidence of neurologic abnormalities--i.e., unexplained, diffuse encephalopathy manifested clinically as chronic progressive dementia. To define the role of human T-cell lymphotropic virus Type III (HTLV-III), the etiologic agent of AIDS, in the pathogenesis of neurologic dysfunction, we compared cerebrospinal fluid and serum from patients with neurologic symptoms associated with AIDS and the AIDS-related complex for the presence of antibodies directed against HTLV-III. Antibodies directed against HTLV-III antigens were detected by four immunologic tests: a fixed-cell immunofluorescence assay, an enzyme-linked immunosorbent assay, immunoblots of viral lysates, and immunoprecipitation of cellular lysates. All patients were seropositive, and 22 of 23 (96 per cent) had HTLV-III-specific antibodies in their cerebrospinal fluid. Unique oligoclonal IgG bands were detected in the cerebrospinal fluid, and the rate of IgG synthesis within the blood-brain barrier was elevated. In eight of nine patients tested, the enzyme-linked immunosorbent assay showed that the percentage of HTLV-III-specific IgG in cerebrospinal fluid was higher than in serum, suggesting that HTLV-III infection of neurologic tissue occurs in the majority of patients with neurologic disease associated with AIDS or its related complex.     

Detection of HTLV-III antibody in cerebrospinal fluid from patients with AIDS and pre-AIDS with the use of a commercial test system

Sixty-one paired sera and cerebrospinal fluid (CSF) specimens were tested for the presence of human T-cell lymphotropic virus/lymphadenopathy-associated virus (HTLV-III) antibodies. Of 16 sera negative for HTLV-III antibodies by enzyme-linked immunosorbent assay (ELISA), the corresponding CSF results were also negative. Forty-two of 45 (93%) CSF specimens tested (in which the corresponding sera were HTLV-III antibody positive) were positive by ELISA. Western blot analysis (WB) of 24 paired specimens, as well as retrospectively acquired case histories, confirmed the results obtained by ELISA.     

Antibodies to Human T Lymphotropic Virus Type III Demonstrated by a Dot Immunobinding Assay

A dot immunobinding (DIB) technique was applied to the demonstration of antibodies to human T-lymphotropic virus type III (HTLV-III). By this technique IgG antibodies to HTLV-III were demonstrated in six of six Swedish patients with acquired immune deficiency syndrome and in 37 of 39 (95%) Swedish homosexual men with persistent generalized lymphadenopathy but in none of 50 Swedish healthy blood donors. Findings obtained by this method showed complete concordance with those obtained by an enzyme-linked immunosorbent assay. All sera positive by DIB were also positive by Western blotting. The DIB assay is simple and well suited for screening of sera for HTLV-III antibodies.     

HTLV-III antibody prevalence among young delinquent drug abusers in long-term residential treatment at a north-German drug clinic

Summary To date, long-term intravenous drug users studied at regional European centers of illicit drug use have had a high number of LAV/HTLV-III infections. Among 200 patients remanded by court or referred from prison to a special clinic in northern Germany for young delinquent drug abusers, 26 (17%) of 157 IV drug abusers were HTLV-III seropositive. With 40% seropositive, female patients showed a significantly higher prevalence of HTLV-III infection than males. The results of longitudinal serological, immunological, and clinical observations over periods of 12 months and 2–3 years indicate that under conditions of continuous medical surveillance, sound preventive counseling, and steady therapeutic care during long-term coeducative residential treatment of drug abusers, neither detectable HTLV-III transmission nor definite progression of HTLV-III induced impairment of immune regulatory functions must ensue.Abbreviations AIDS acquired immune deficiency syndrome - CDC Centers for Disease Control - HTLV-III human T-lymphotropic virus type III - IV intravenous - LAV lymphadenopathy-associated virus     

Coxsackie B virus-specific IgM responses in coronary care unit patients

An enzyme-linked immunosorbent assay (ELISA) test using polyvalent antigens and antisera was used to detect Coxsackie B virus-specific IgM responses in 329 patients admitted to the Coronary Care Unit, Wellington Hospital, New Zealand over a 12-month period. The sera of 30 of 153 (19.6%) patients with acute myocardial infarction (AMI), 16 of 98 (18.4%) with chest pain, and 7 of 46 (15.2%) patients with arrhythmia were positive for Coxsackie B virus-specific IgM. Four of 12 (25%) patients with heart failure were also positive. Over the same period, 178 sex- and age-matched normal blood donors were also studied. Eleven of 178 (6.2%) matched blood donors were positive for Coxsackie B virus-specific IgM. The rates of occurrence of Coxsackie B virus-specific IgM in patients with AMI and in a group of matched controls showed a significant difference (chi 2 = 5.64, p = 0.02).     

Antiviral antibodies in the sera of homosexual men: correlation with their lifestyle and drug usage

Healthy homosexual men between the ages of 21 and 65 years, from the Washington, DC (n = 162), and New York City (n = 89) areas, were studied for antibodies in the serum against cytomegalovirus (CMV), herpes simplex virus (HSV) types 1 and 2, and Epstein Barr virus (EBV) viral capsid antigen (VCA). CMV-specific antibodies were assayed by enzyme-linked immunosorbent assay (ELISA), anti-HSV-1 and -2 antibodies were measured by indirect hemagglutination (IHA), and antibodies to EBV VCA were measured by the immunofluorescence assay. Antibodies to human T lymphotrophic virus III (HTLV-III) were detected by ELISA and Western blot procedures. T lymphocytes were enumerated using OKT4 monoclonal antibody. Healthy male volunteer blood donors (n = 90) matched for age range and race proportions were used as controls. The percentage of seropositive individuals in the homosexual group was higher (90-98%) for all the viruses tested than in the control group (47-87%). Comparisons of the geometric mean titers, expressed as reciprocal serum dilutions, of seropositive individuals in homosexual (H) vs control (C) group were as follows: CMV-IgG (ELISA) H = 1:794, C = 1:68; HSV-1 (IHA) H = 1:248, C = 1:14; HSV-2 (IHA) H = 1:56, C = 1:17; EBV-VCA (IFA) H = 1:385, C = 1:131. The homosexual group also showed a higher frequency of individuals with elevated titers than the control group. The CMV IgM antibody was prevalent in 17.7% of the homosexual group and 5% of the control group; arithmetic means for ELISA values for CMV IgM were 0.207 for the homosexual group and 0.05 for the control group. In the homosexual group, the anti-CMV antibody titers increased with age (P = 0.01) and with numbers of sex partners (P = 0.06). Both anti-HSV-1 and anti-HSV-2 antibodies correlated with the number of sex partners (P = 0.04 and P = 0.05, respectively). Neither age nor partner number correlated with response to EBV, and no particular sex act was related to the EBV VCA titer level. HTLV-III seropositivity was associated with higher herpes virus group antibody titers, probably because of life style cofactors. Among the HTLV-III-seropositive subjects, those with less than or equal to 400 T-helper lymphocytes/mm3 had lower antibody titers than those with greater than 400 T-helper lymphocytes/mm3 counts, suggesting an impaired immune response secondary to immunosuppression.     

Lymphadenopathy and antibodies to HTLV-III in homosexual men

Summary The study provides information on the epidemiology of HTLV-III infection and the lymphadenopathy syndrome (LAP) in 374 German homosexual men. Sexual contacts in the USA and rectal enemas before receptive anal intercourse are the main risk factors associated with virus transmission. HTLV-III seropositivity is significantly correlated with LAP. Prominent clinical signs are infreqquent. Immunological and haematological abnormalities are prevalent, and the retrovirus infection is frequently associated with serological markers of other viruses (hepatitis B, herpes group viruses). Lymphadenopathy as a manifestation of HTLV-III infection is discussed within the context of AIDS-related disorders.Abbreviations AIDS Acquired Immunodeficiency Syndrome - CMV Cytomegalovirus - EBV Epstein-Barr virus - HBV Hepatitis B virus - HTLV-III Human leukaemia retrovirus type III - LAP Lymphadenopathy     

Evaluation of three commercial screening tests for AIDS virus antibodies

Three commercial enzyme-linked immunosorbent assays (ELISA) for acquired immune deficiency syndrome (AIDS) virus antibodies were evaluated using serum that had been characterized by an ELISA and western blot procedure developed at the University of California at Davis (UCD). Each of the commercial tests was more specific than the UCD ELISA, but the UCD ELISA was more sensitive in the detection of sera that lacked reactivity by western blot to the envelope glycoprotein (gp-41). The HTLV-III Bio-EnzaBead (Litton Bionetics, Charleston, SC) was less sensitive and specific than the Abbott HTLV-III EIA (Abbott Laboratories, North Chicago, IL) or the Virgo HTLV-III ELISA (Electro-Nucleonics Inc., Columbia, MD). Overall, 22.9% (57 of 250) and 51.0% of sera that were repeatedly (X2) positive by commercial screening kits tested at blood donor centers and clinical laboratories, respectively, were confirmed by western blot. These results indicate that the screening assays for AIDS virus antibodies are not equal in performance and that positive screening test results must be confirmed by a more specific test like western blot before results are released.     

Risk of nosocomial infection with human T-cell lymphotropic virus III (HTLV-III)

Infection with human T-cell lymphotropic virus III (HTLV-III) is closely linked to the acquired immunodeficiency syndrome (AIDS). We evaluated the risk of nosocomial infection with HTLV-III by testing for antibodies to HTLV-III among hospital employees, including victims of needle-stick exposure, endoscopists, pathologists, and laboratory workers. Assays for antibody against the virus were performed by enzyme-linked immunosorbent assay and electrophoretic (Western blot) techniques. Although all 22 of our patients with AIDS and 6 of 7 with AIDS-related complex were found to have antibodies to HTLV-III when both assays were employed, none of the 85 employees with nosocomial exposure to specimens from patients with AIDS were positive for HTLV-III antibody. These studies must be regarded as preliminary, but they suggest that when current hospital isolation procedures are employed, the risk of nosocomial transmission of HTLV-III is low.     

Testing for antibodies to AIDS-associated retrovirus (HTLV-III/LAV) by indirect fixed cell immunofluorescence: specificity, sensitivity, and applications

Seropositivity to the AIDS-associated retrovirus, HTLV-III/LAV, has profound implications. Simple and reliable tests are needed to detect such antibodies. A rapid, sensitive indirect immunofluorescence assay (IFA) on acetone-fixed virus-producing CEM/LAV-N1 cells was adapted for detection of human antibodies to HTLV-III/LAV. Specific and nonspecific patterns of of immunofluorescent reactivity were easily distinguished, and results paralleled those obtained by Western blotting and radioimmunoprecipitation (RIP), indicating that there is no need to confirm IFA positivity. In contrast, the commercial enzyme-linked immunosorbent assay (ELISA) was less reliable. False positives occurred with sera from seven hemophiliacs that were negative on Western blots, and false-negative reactions were observed on two occasions. These involved low-titer AIDS-patients' sera that were positive on Western blots, and from one of which virus was successfully isolated. Our results emphasize the requirement for confirmatory assays when the ELISA test is used for primary screening of sera for antibodies to HTLV-III/LAV. The IFA method is especially well-suited to quantitative analysis of serum antibody levels. Our data suggest that serum antibody titers rise as disease progression occurs, ultimately falling as severe complications ensue. It is suggested that in laboratories where the demand for HTLV-III/LAV antibody testing is not excessive (1,000-2,000 sera/month), IFA could serve as the only serological assay for both screening and epidemiological purposes.     

A glycopolypeptide (gp 100) is the main antigen detected by HTLV-III antisera

Sera from homosexuals and hemophiliacs in Germany were examined for antibodies to human T-lymphotropic retrovirus type III (HTLV-III) by an enzyme-linked immunosorbent assay (ELISA) against purified virus. ELISA positive sera were used to search by immunoprecipitation for HTLV-III related antigens in a persistently infected human T-cell line. A glycopolypeptide with a mol. wt. of 100 000 was regularly recognized by all positive sera. In analogy to glycosylation and high antigenicity of envelope polypeptides of other mammalian retroviruses, gp100 seemed to be related to the env gen. Two polypeptides with mol. wts. of 24 000 and 22 000 probably representing viral core polypeptides were additionally detected by sera with high ELISA titers.     

Immune functions in homosexual men with antibodies to HTLV-III in Finland.

The occurrence of HTLV-III antibodies in a voluntary group of 175 homosexual men in a low risk AIDS area was studied, and the findings were correlated to clinical, virological, immunological and lifestyle parameters. Fifteen of 175 men had HTLV-III antibodies; two of these had AIDS, five had LAS and two had enlarged lymph nodes. In the HTLV-III antibody negative group, no signs of AIDS or pre-AIDS were seen during a 10 month follow-up. In HTLV-III antibody positive individuals, low TH/TS ratio was mainly due to decreased number of TH cells. Most HTLV-III antibody positive cases had low responses to a specific antigen, PPD, while responses to the mitogens PHA and PWM were only slightly affected. In HTLV-III antibody negative cases, 13% had a low TH/TS ratio, mostly due to elevation of TS cells. In this group, mitogen and antigen responses were normal or only slightly affected. The results reinforce the causal relationship between HTLV-III and AIDS and suggest that the cells primarily affected by the virus infection are TH cells, responsible for antigen specific responses. Longitudinal studies are required to find out, what is the relationship of immune response to the development of clinical AIDS in HTLV-III infected individuals.     

Subclass distribution of rubella virus-specific immunoglobulin G

An enzyme-linked immunosorbent assay was used to study the subclass distribution of rubella virus-specific immunoglobulin G (IgG) in 97 serum samples from healthy donors and from patients with recent or remote rubella infections. Plastic beads coated with rubella antigen were incubated with test serum and then with monoclonal antibodies to the four human subclass of IgG. Rubella virus-specific IgG1 was present in all serum samples containing rubella virus-specific IgG antibodies. Rubella virus-specific IgG2 was present in 1 of 35 samples from healthy donors that also contained specific IgG1. Rubella virus-specific IgG3 was found in serum samples from patients with recent rubella infections but had disappeared by 6 months after the onset of symptoms. Rubella virus-specific IgG4 was found in low amounts in 7 of 35 samples from healthy immune donors. Of 20 serum samples that were negative by other serological techniques, 8 gave absorbances above cutoff levels in the assays for rubella virus-specific total IgG and IgG1. In 1 of 20 serum samples, the assays for total IgG and IgG2 were positive. High absorbance in the assay for rubella virus-specific IgG4 was found in one serum. This serum was negative in all other assays for rubella virus-specific antibodies.     

Recombinant Protein-44-Based Class-Specific Enzyme-Linked Immunosorbent Assays for Serologic Diagnosis of Human Granulocytic Ehrlichiosis

Recombinant protein 44, expressed and purified as a maltose-binding protein fusion peptide of the human granulocytic ehrlichiosis (HGE) agent (Ehrlichia phagocytophila genogroup), was used as antigen in enzyme-linked immunosorbent assays (ELISAs) to detect total antibodies, immunoglobulin (Ig) M antibodies, and IgG antibodies. Of the 67 human sera obtained from 64 HGE patients 3-5 weeks after the onset of illness and confirmed as having total immunoglobulins to whole-cell antigen by indirect fluorescent antibody analyses, 63 were positive in a polyvalent ELISA. Fifty-six and 61 sera had IgM or IgG antibodies, respectively. Fifty sera had both IgM and IgG antibodies. In specificity tests of 110 sera, one serum sample from a patient who had Lyme borreliosis reacted to the protein 44 antigen in the analysis for IgM antibody (specificity, 99%). There were no false-positive results in an ELISA for IgG antibodies. With their high sensitivity and specificity, class-specific ELISAs can be used in conjunction with indirect fluorescent antibody analyses or immunoblotting methods to help diagnose human granulocytic ehrlichiosis.     

Use of monoclonal antibodies to detect antigens of Toxoplasma gondii in serum and other body fluids.

Monoclonal antibodies to Toxoplasma gondii were used in an enzyme-linked immunosorbent assay to detect antigens of the parasite in toxoplasma lysate, in peritoneal fluid of mice, and in sera from humans acutely infected with T. gondii. Four of the six monoclonal antibodies were able to detect antigens of toxoplasma in these specimens. Control sera from individuals not infected with T. gondii and from individuals chronically infected with the parasite were negative in the enzyme-linked immunosorbent assay. Sera from individuals not infected with T. gondii but with positive titers from rheumatoid factor were also negative; 2 or 10 sera from individuals not infected with T. gondii but with positive titers for antinuclear antibodies reacted with the monoclonal antibodies. When the results of the enzyme-linked immunosorbent assay with monoclonal antibodies and with the F(ab)2 fraction of an immunoglobulin G from a rabbit infected with T. gondii were compared, it was noted that the F(ab)2 was more active in detecting parasite antigens than were the monoclonal antibodies. Thus, although monoclonal antibodies can be used to detect antigens of T. gondii in sera and other body fluids, polyvalent antibody (such as the F(ab)2 fraction) appears to be more satisfactory for this purpose.     

Serologic diagnosis of canine and equine borreliosis: use of recombinant antigens in enzyme-linked immunosorbent assays.

Serum samples from dogs and equids suspected of having canine or equine borreliosis, respectively, were analyzed in polyvalent enzyme-linked immunosorbent assays (ELISAs) with whole-cell or recombinant antigens of Borrelia burgdorferi sensu stricto. Purified preparations of recombinant antigens included outer surface protein A (OspA), OspB, OspC, OspE, OspF, and p41-G (a fragment of flagellin). Of the 36 dog sera that reacted positively to whole-cell antigen, 32 (88.9%) contained antibodies to one or more recombinant antigens. Reactivities to OspF (88.9% positive) and p41-G (75% positive) were most prevalent. In analyses of 30 equid sera positive in an ELISA with whole cells, 24 (80%) contained antibodies to one or more recombinant antigens. Seropositivities in ELISAs with p41-G (50% positive) and OspF (46.7% positive) were more than twofold greater than in ELISAs with OspA, OspB, or OspC (10 to 20% positive). In parallel tests of eight canine and three equine sera, there was good agreement in results of Western blot (immunoblot) analyses and ELISAs. Although dog and equid sera with antibodies to whole-cell B. burgdorferi frequently reacted positively to one or more recombinant antigens, the inclusion of OspF and p41-G antigens in ELISAs was most useful in the serologic diagnosis of canine and equine borreliosis.