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Biomedical subjects

A L Cunningham

Publications and source records attributed to A L Cunningham.

At least 127 records · Page 7Linked to original sources

Roseola infantum and other syndromes associated with acute HHV6 infection.

Eight cases of acute human herpesvirus type 6 (HHV6) infection in infants were diagnosed serologically by the demonstration of IgM anti-HHV6 (8/8) and a significant change in total anti-HHV6 antibody titre (6/8). Four infants were sufficiently ill to require admission to hospital and further investigations: one with encephalitis and three with gross hepatosplenomegaly, two of whom had evidence of simultaneous infection with another herpes-virus. The remaining four infants had an illness compatible with roseola infantum, although this diagnosis had not been made clinically. Sera from two of those infants with rash had been sent for analysis to exclude rubella because the infants' mothers were pregnant. The other two had received antibiotics when febrile, and the subsequent appearance of the roseola rash had raised the possibility of antibiotic allergy. The data suggest that there are clinical syndromes in addition to roseola infantum associated with the presence of IgM anti-HHV6, in which serological screening for evidence of acute HHV6 infection may be useful.

Antibodies, Viral↗

Should all pregnant women be screened for hepatitis B surface antigen?

One thousand, one hundred and ninety-three pregnant women were screened for hepatitis B surface antigen (HBsAg) carriage after their histories had been reviewed to assess their risk status. In this prospective study, carrier rates were examined according to conventional risk criteria and also according to extended criteria that included the country of birth of the patient's parents. Twenty-six (2.2%) patients were found to be HBsAg seropositive, and one of these patients showed no identifiable risk factors. In four hepatitis B virus carriers, the only risk factor was that the patients' parents had been born in a country that was classified as high risk. Thus, five (19%) of 26 patients would not have been identified by means of previously-accepted screening procedures. Four hundred and forty-two (37%) patients showed at least one conventional risk factor and 558 (47%) patients showed at least one risk factor by our extended criteria. Given the high costs to the community of chronic hepatitis B virus carriage, it was concluded that the screening of all antenatal clinic patients for the presence of HBsAg is cost effective.

Australia↗

Rapid diagnosis of varicella-zoster virus infection with a monoclonal antibody based direct immunofluorescence technique.

Diagnosis of varicella-zoster virus (VZV) infection in immunocompromised patients is difficult because of the frequent atypical appearance. Accurate and early diagnosis is important to allow rapid commencement of antiviral chemotherapy, with consequent improvement in antiviral efficacy. A monoclonal based direct immunofluorescence antibody technique (VZV IFA) was assessed in parallel with viral culture in 56 patients with suspected VZV infection. A subgroup of 17 patients from this group with classical dermatomal herpes zoster all had positive VZV IFA tests. Only 6 patients (35%) were positive on viral culture. None of the 15 patients with proven herpes simplex virus infection had a positive VZV IFA, nor did any patient with positive VZV viral culture have a negative VZV IFA. The VZV IFA test is a rapid and sensitive technique for detecting infection with VZV.

Antibodies, Monoclonal↗

HIV-1 antibody testing strategy: evaluation of ELISA screening and western blot profiles in a mixed low risk/high risk patient population.

The Westmead HIV-1 antibody testing strategy showed that, regardless of ELISA screening kit manufacturer, sera which were repeatedly positive by two ELISA screening assays (one indirect and the other competitive format) had a 97-98% chance of being confirmed positive by Western blot for HIV-1 antibody or a less than 3% chance of either being identified as a seroconverter (1%) or a late stage AIDS patient (1.2%). Sera which were discordant by two ELISA screening assays had a less than 4% chance of either being confirmed positive by Western blot (2.5%) or identified as a seroconverter (1.3%). The incidence of non-specific indeterminate Western blot profiles were shown to be inversely proportional to the specificity of the ELISA screening kits used. The use of a recombinant envelope ELISA was able to confirm the viral specificity of HIV-1 envelope bands (gp160, 120 or 41) on Western blot. Guidelines suggested by the Australian National HIV Reference Laboratory, Fairfield Hospital, Melbourne, which categorized indeterminate or typical Western blot profiles into four reaction groups were found to be useful for the interpretation of Western blot patterns. A Western blot profile which is reactive for HIV-1 viral glycoproteins (gp160, 120 and 41) alone or in combination with not more than two other viral proteins (Indeterminate Group 4) and which is confirmed viral envelope specific by a recombinant envelope ELISA can be used as a predictor of seroconversion.

AIDS Serodiagnosis↗

Variations in CD4 expression by human monocytes and macrophages and their relationships to infection with the human immunodeficiency virus.

The expression of CD4 antigen on the surface of LeuM3-positive human blood monocytes was found to be variable with 65 to 90% of cells from 46 normal human volunteers being positive by dual staining flow cytometry. When monocytes adhered to plastic (but not when cultured on Teflon), a marked decrease in CD4 expression was observed between 1 and 24 h post-adherence. CD4 expression could not be detected in macrophages adhered to plastic for 5 days by using four anti-CD4 monoclonal antibodies in flow cytometry or direct immunofluorescence. Conversely an increasing proportion of adherent cells expressed LeuM3 and OKM5 surface antigens over the 5 days. CD4 mRNA levels were measured by slot-blot and Northern hybridization, and total cellular CD4 protein levels by immunoprecipitation. Both cellular mRNA and CD4 levels remained constant throughout the 5 day period but membrane CD4 protein levels were greatly reduced indicating that the down-regulation of CD4 was post-translational. Infection with two of six fresh human immunodeficiency virus (HIV) isolates showed different kinetic patterns when tested on purified monocytes recently adhered to plastic and macrophages adherent for 5 days. HIV antigen and reverse transcriptase levels in infected monocyte cultures remained high for 3 to 4 weeks before detachment and necrosis of the cells occurred. Infection of macrophages generated much lower levels of antigen and reverse transcriptase which declined to very low or undetectable levels over 2 weeks, leaving persisting viable macrophages. One week after infection HIV nucleic acid was detected in 69 +/- 7% of monocytes and 6 +/- 3% of macrophages by in situ hybridization. Blocking experiments with anti-Leu3a monoclonal antibody suggested that HIV infection of 5 day adherent macrophages occurred mainly by a mechanism other than binding to CD4.

CD4 Antigens↗

Rapid diagnosis of acute Epstein-Barr virus infection by an indirect enzyme-linked immunosorbent assay for specific immunoglobulin M (IgM) antibody without rheumatoid factor and specific IgG interference.

An indirect enzyme-linked immunosorbent assay (ELISA) for detection of Epstein-Barr virus-specific immunoglobulin M (IgM) antibody was developed with commercial reagents. Sera containing rheumatoid factor (RF) (as little as 0.5 IU/ml) coupled with specific IgG resulted in false-positives in the ELISA. This interference was eliminated by the use of anti-human IgG antibodies to remove RF and IgG. Thus, pathogen-specific IgG complexes to which IgM-RF could be bound during the subsequent test were inhibited, and competition between specific IgG and IgM was also prevented. Of the 1,672 serum specimens tested, 353 were found to be Epstein-Barr virus IgM antibody positive by indirect immunofluorescence (IF). Compared with the IF test, the ELISA showed 96.6% sensitivity, 99.7% specificity, and 99% accuracy. Further evidence indicated that most of the 12 ELISA false-negatives were IF false-positives. There was a linear correlation between mean ELISA values and increasing IF titers (r = 0.96). However, the IF test has the disadvantages that it lacks automated reading and requires considerable technical expertise, both of which restrict the range of laboratories performing the test. The indirect ELISA has the advantages that it is simple and rapid and can be automated. All the reagents used in this assay are commercially available, have been prestandardized, and are stable.

Adolescent↗

Role of keratinocytes in human recurrent herpetic lesions. Ability to present herpes simplex virus antigen and act as targets for T lymphocyte cytotoxicity in vitro.

In human recurrent herpetic lesions epidermal keratinocytes are induced to express HLA class II (DR) antigens. Keratinocytes derived from human split skin and cultured in vitro were induced to express HLA-DR but not -DQ antigens with IFN gamma preparations. These stimulated keratinocytes presented herpes simplex antigen directly to autologous blood-derived T lymphocytes in four of four subjects (stimulation indices: 1.5-2.7), suggesting that keratinocytes may have an accessory herpes simplex virus (HSV) antigen-presenting role in addition to the Langerhans cells and macrophages in herpetic skin lesions. Blood mononuclear cells from eight herpes simplex seropositive subjects which were activated in vitro by HSV antigen for 6 d showed cytotoxicity specific for HSV in infected autologous keratinocytes. This was significantly increased by prestimulation with IFN gamma (51-56% to 83-85%). In four of eight patients some cytotoxicity also occurred against uninfected, IFN gamma-stimulated keratinocytes. Lymphocyte subset analysis showed that cytotoxicity against HSV-infected, IFN gamma-stimulated keratinocyte targets was mediated by both CD3+ T lymphocytes and Leu 11b+ natural killer cells. T lymphocyte cytotoxicity was mediated by both CD4+ and CD8+ T lymphocytes, suggesting a cytotoxic role for the activated CD4+ lymphocytes that initially predominate in herpetic lesions.

Antigens, Differentiation, T-Lymphocyte↗

Immune complexes and Ross River virus disease (epidemic polyarthritis).

Immune complexes were sought in serum and synovial fluid in Ross River virus disease (epidemic polyarthritis). Multiple samples from 15 patients showing varied degrees of disease activity over a 3 month period were analysed for their content of complement components C3 and C4, and for C1q solid-phase and Raji cell binding activity. Levels of C3 and C1q binding activity were normal. C4 and Raji cell binding activity were normal except for three high levels of Raji cell binding, of which two were accompanied by low levels of C4, with normal C3 and C1q binding. Synovial fluid showed anomalous Raji cell reactivity of uncertain significance. Conglutinin solid-phase binding activity and IgG rheumatoid factor were compared in the serum of 20 patients during active disease and after recovery. The results were identical and within the normal range in both phases. One patient developed IgM rheumatoid factor in a low titre late in his illness. Although these findings do not entirely exclude a role for immune complexes formed at the onset in the circulation or tissues, it is concluded from this and other evidence that circulating complexes are not commonly responsible for the persistence of syndromes in this disease.

Antigen-Antibody Complex↗

Gastrointestinal viral infections in homosexual men who were symptomatic and seropositive for human immunodeficiency virus.

Gastrointestinal viruses, predominantly rotaviruses and adenoviruses, were detected by enzyme-linked immunosorbent assay, electron microscopy, or cell culture in greater than 50% of two groups of homosexual men with symptomatic human immunodeficiency virus (HIV) infection, who did (54%) or did not (50%) have diarrhea. Lower detection rates were observed in HIV-seronegative (15%) and asymptomatic HIV-seropositive (16%) men. In the patients with diarrhea, 95% of the isolates of virus were found in the most immunosuppressed patients, those patients with AIDS-related complex or opportunistic infections associated with AIDS. High excretion rates of these viruses are probably associated with both anal-oral transmission and immunosuppression. These viruses apparently cause acute episodes or relapses of diarrhea in some patients but may be co-pathogens or noncontributory to chronic diarrhea in others.

AIDS-Related Complex↗

Evaluation of rubella immune status by three commercial enzyme-linked immunosorbent assays.

Three commercial indirect enzyme-linked immunosorbent assays (ELISAs) (Enzygnost-Rubella, RUBELISA, and ORTHO Rubella) were evaluated for the determination of immune status by testing 1,090 serum specimens, 410 of which were from nonimmune patients. In comparison with the standard reference technique, the hemagglutination inhibition (HAI) test, the sensitivities of ORTHO Rubella (100%) and Enzygnost-Rubella (99.26%) were excellent, whereas the sensitivity of RUBELISA (95.60%) was marginally lower because of the inability of this assay to detect antibody in 22% of the serum specimens with HAI titers of 10 and 11% of sera with HAI titers of 20. The specificity of all three systems was greater than 97%. There was a linear correlation between mean ELISA values and increasing HAI titers (r greater than or equal to 0.94). Both ORTHO Rubella and Enzygnost-Rubella were shown to be suitable replacements for the HAI test, provided that an equivocal zone is incorporated in the ORTHO system and only unheated sera are used in the Enzygnost system.

Antibodies, Viral↗

Glomerulonephritis in the acute phase of Ross River virus disease (epidemic polyarthritis).

Hematuria and proteinuria were detected at the peak of symptoms in a case of Ross River virus (RRV) disease. No other infective cause was identified. A renal biopsy 28 days after the onset of nephritis showed mild mesangial proliferative changes and one segmental sclerotic lesion. Immunofluorescence showed widespread linear deposition of IgG in glomerular capillary walls with similar but weak staining for IgM, complement (C3) and fibrinogen; granular deposits of IgM and C3 in several arterioles; and IgM in a few mesangial cells. No electron-dense deposits were detected, nor was RRV antigen found in the renal tissue. Anti-glomerular basement membrane antibodies were not detected in the serum. Recovery from the renal disturbance was complete within three months although rheumatic symptoms persisted for 30 months.

Acute Disease↗