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Biomedical subjects

J R Pattison

Publications and source records attributed to J R Pattison.

At least 37 records · Page 2Linked to original sources

The development and use of an antibody capture radioimmunoassay for specific IgM to a human parvovirus-like agent.

An IgM-antibody capture radioimmunoassay (MACRIA) was developed for the detection of IgM antibody specific for the human parvovirus-like agent B19. Diagnosis of infection with this agent by either antigen detection or antibody seroconversion had been made by counter-current immunoelectrophoresis (CIE) in 18 cases of aplastic crisis occurring in children with homozygous sickle-cell disease. The MACRIA described here gave positive results in 17 of these 18 cases; in the remaining case only an acute specimen taken from the patient during viraemia and late convalescent specimens taken 184 and 247 days after onset of illness were available. The test was used to investigate 20 further cases of aplastic crisis in which neither viral antigen nor antibody seroconversion could be detected by CIE. Detection of virus-specific IgM permitted diagnosis of infection with this parvovirus-like agent in 17 of these cases. In the remaining three cases only single serum specimens taken late in convalescence, 82 days or more after the onset of symptoms, were available. In addition to these 34 cases of aplastic crisis in which primary infection with this agent was diagnosed by MACRIA, seven cases of apparent 'silent' infection detected by CIE were investigated. The test permitted the discrimination between primary infection and re-exposure to the virus in six of these patients. The use of this assay has added a considerable weight of evidence implicating primary infection with this parvovirus-like agent as an important cause of aplastic crisis in children with sickle-cell disease. Furthermore, MACRIA permits diagnosis of infection when only single serum specimens taken up to ten weeks after infection are available. Thus the use of this test will significantly facilitate the investigation of other clinical syndromes of presumptive infectious aetiology.

Adult↗

Occurrence of infection with a parvovirus-like agent in children with sickle cell anaemia during a two-year period.

The occurrence of infection with a parvovirus-like agent during the period April 1979-May 1981 in children attending a single sickle cell clinic in London was investigated. Virus was detected in serum by counter-current immunoelectrophoresis (CIE) and immunoelectron microscopy (IEM). Viral antibody was detected by CIE and specific IgM antibody by an IgM-antibody capture assay. Of the 68 children studied nine presented in aplastic crisis and evidence of infection with the parvovirus-like agent at the time of the crisis was found in all nine. Eighteen of the other children were antibody-positive at some time during the study. In 11 children there was no evidence of recent infection; however, two of these had a history of aplastic crisis in previous years. The other seven seroconverted during the course of the study but did not show any haematological effects. Five of these had a primary infection, one appeared to have reinfection and in the seventh there were insufficient data to distinguish between the two. Possible explanations for the difference between those presenting with aplastic crisis and those with asymptomatic seroconversion are discussed.

Anemia, Sickle Cell↗

Outbreak of aplastic crises in sickle cell anaemia associated with parvovirus-like agent.

Since 1952, 112 children with sickle cell anaemia (SCA) in Jamaica have had an aplastic crisis. Outbreaks occurred in 1956, 1960, 1065-67, 1971-73, and 1979-80. Most cases occurred in children under 10 years of age, and an aplastic crisis in a patient over the age of 15 years is rare. There were 38 cases in 1979-80 and stored serum specimens from 28 of these were available for virus studies. Evidence for infection with a parvovirus-like agent was found in 24 of these 28 cases. Viral antigen was detected in 2 patients, both of whom demonstrated seroconversion. Seroconversion during 1980 was detected in a further 7, increasing amounts of antibody during the convalescent period were found in 5, antibody was found in 2 of 4 patients from whom only an acute phase specimen was available and the remaining 10 were antibody positive in the only convalescent phase sample available for testing. Antibody was found in 4 of 94 controls with the SS genotype (in retrospect 2 of these may have had an aplastic crisis) and in 17% of 48 controls with a normal haemoglobin (AA) genotype. The results accord with the possibility that the parvovirus-like agent is the principal cause of aplastic crisis in SCA.

Adolescent↗

Outcome of asymptomatic infection with rubella virus during pregnancy.

We have tried to detect prenatal infection in 34 infants whose mothers were re-infected with rubella virus during pregnancy and in six infants whose mothers had primary subclinical rubella during pregnancy. Two methods of assessment were used: first, serum obtained soon after birth was tested for IgM antibody; secondly, serum obtained after the age of 8 months was tested for specific IgG. The 34 women with re-infections had increases in IgG antibody titre but no IgM response. No evidence of prenatal infection was found in 33 of their 34 infants. One infant was found to have IgG antibody at the age of 11 months. This infant was IgM-negative at birth and had a rubelliform rash at the age of 5 1/2 months; it therefore probably contracted post- rather than pre-natal infection. Fetal infection from maternal re-infection during pregnancy is probably rare. The six women with primary subclinical rubella produced both IgG and IgM classes of antibody. Three of their six infants showed serological evidence of intrauterine infection. One, infected when its mother was 8 weeks pregnant, had clinical evidence of congenital rubella. Primary subclinical rubella during pregnancy therefore carries a significant risk of fetal infection. Because of the difference in outcome, great care should be taken to distinguish between primary infection and re-infection when investigating symptomless increases in antibody titre after contact with rubella during pregnancy.

Antibodies, Viral↗

Fetal infection resulting from maternal rubella after the first trimester of pregnancy.

We have tried to measure the incidence of prenatal infection in 304 infants whose mothers had had rubella at various times after the first 12 weeks of pregnancy. Two methods of assessment were used: first, serum obtained soon after birth was tested for specific IgM antibody; secondly, serum obtained after the age of eight months was tested for specific IgG. When maternal rubella occurred 12-16 weeks after the last menstrual period specific IgM antibody was detected in 28 out of 50 infants (56%). The proportion fell progressively to 12% after maternal rubella at 24-28 weeks, rose to 19% after rubella at 28-36 weeks and then to 58% when the illness occurred during the last month of pregnancy. In all, IgM antibody was detected in 77 out of 260 infants (29%). The fetus can thus be infected at any time during the second and third trimesters of pregnancy, but the risk varies at different stages.The figures for the prevalence of IgG antibody were greater throughout, because some infants had IgG who had previously lacked specific IgM. After maternal rubella at 12-16 weeks IgG antibody persisted in 22 out of 31 infants (71%). The proportion fell to 28% after rubella at 24-28 weeks and then increased progressively to 94% after rubella during the last month. In all, IgG antibody persisted in 94 out of 190 infants (49%). The true rate of fetal infection probably lies between the rates estimated from the presence of IgM antibody and the subsequent prevalence of IgG.Infants whose mothers had rubella at any time during pregnancy should be examined regularly for possible evidence of damage.

Antibodies, Viral↗

The role of host responses in the recovery of mice from Sendai virus infection.

The antiviral responses in mice to intranasal inoculation with Sendai virus are described. To investigate the relative importance of the humoral, cell-mediated and interferon responses, the pathogenesis of this infection was studied in animals which were immunocompetent, T cell-deprived or immunosuppressed with cyclophosphamide. Treatment with cyclophosphamide converted the mild, self-limiting infection observed in immunocompetent mice into a severe and frequently lethal pneumonic disease. This was associated with an enhanced interferon response but no detectable antibody or cell-mediated immune response. T cell-deprived mice suffer an infection of intermediate severity associated with an increased interferon response, a normal humoral immune response and no cell-mediated immune response. The implications of these results in relation to the role of the antiviral responses in recovery from Sendai virus infection are discussed.

Animals↗

Rubella-specific IgM detection using Sephacryl S-300 gel filtration.

Sephacryl S-300 has been compared with Sephadex G-200 in the fractionation of sera for the detection of rubella-specific IgM. No difference in sensitivity was found with sera that had low titres of rubella-specific IgM. Sephadex G-200 was apparently more sensitive with some sera which had high titres of rubella-specific IgM due to the contribution of rubella-specific IgA. Sephacryl S-300 offers considerable advantages in ease and speed of preparation, and as it is a more rigid gel higher flow rates can be obtained. Sera may be fractionated in 2.5 hours on Sephacryl S-300 compared with 7-10 hours on Sephadex G-200.

Antibody Specificity↗

Comparison of immunofluorescence and radioimmunoassay for detecting IgM antibody in infants with the congenital rubella syndrome.

Immunofluorescence (IF) and radioimmunoassay (RIA) have been compared as methods for detecting IgM antibody in 124 infants with confirmed or suspected congenital rubella. IF was used to test sucrose density gradient fractions and RIA to test fractions and whole serum. When fractions were tested IF and RIA were equally specific and distinguished clearly between IgM and IgG, but RIA was the more sensitive method. The RIA titre in whole serum was always greater than in the peak IgM fraction and there was no evidence that testing the serum, rather than the fraction, could result in failure to detect IgM. With some sera RIA gave low titres which became negative after absorption with IgG-coated latex beads. The mechanism of this 'false positive' effect, which may have been due to IgM with anti-IgG activity, was not investigated, but if it can be removed by absorption it need not reduce the specificity of the test. During the first 6 months of life IgM antibody was detected by RIA in 30 out of 32 unfractionated sera and by IF in fractions from 28 of these. After the age of 6 months IgM was found progressively less frequently and the greater sensitivity of RIA became a more obvious advantage: 17 out of 60 specimens were positive by RIA and 11 of these were negative by IF. RIA testing of whole serum appears to be an economical, specific and sensitive method for detecting IgM antibody in congenital rubella, of particular value when the titre of antibody is low.

Age Factors↗

A comparison of three tests for rubella antibody screening.

Sera from 1000 women attending antenatal clinics were screened for the presence of rubella antibody by haemagglutination inhibition (HI), single radial haemolysis (SRH), and enzyme-linked immunosorbent assay (ELISA). With 925 sera concordant results were obtained with all three tests. There were 46 false-negative HI tests due to the necessity of allowing for residual non-specific inhibitors. With SRH there was one false positive and three that were negative by SRH but HI- and ELISA-positive. By ELISA 14 sera gave a false-positive result (OD 405 nm greater than or equal to 0.2). Ten of these could be eliminated by taking an OD 405 nm of 0.5 as the threshold but then another 10 sera became false negatives.

Antibodies, Viral↗

The nature of the effector cells of cell-mediated immune responses to Sendai and Kunz virus infections in mice.

The development of virus-specific cell-mediated immune (CMI) responses to Sendai and Kunz virus infections of the respiratory tract of mice is described. In addition a heterotypic cell-mediated immune response develops during each of the virus infections. Cell depletion studies indicate that the effector cells of the virus-specific CMI responses are theta antigen bearing T lymphocytes. Similar studies of the heterotypic responses suggest that 2 cell types are involved, T lymphocytes and NK cells.

Animals↗

The use of high speed gel filtration for the detection of rubella-specific IgM.

Three new gels, Ultrogels AcA 22, AcA 34 and AcA 44 have been compared with Sephadex G-200 for the detection of rubella-specific IgM. Ultrogel AcA 44 did not achieve an adequate separation of IgM and IgG. Ultrogels AcA 22 and AcA 34 gave clear separation of IgM and IgG but only the latter could be operated at flow rates significantly higher than Sephadex G-200. With rubella convalescent sera fractionation with Ultrogel AcA 34 leads to a 2-4-fold loss of sensitivity for the detection of the first peak of rubella HI activity which is associated with recent infection. This is largely due to the occurrence of specific IgA antibody in the first peak of rubella HI activity after Sephadex G-200 fractionation but not after Ultrogel AcA 34 fractionation.

Antibody Specificity↗

The derivation of a minimum immune titre of rubella haemagglutination-inhibition (HI) antibody. A Public Health Laboratory Service collaborative survey.

Ten laboratories collaborated in a study of minimum immune titre (MIT) of rubella haemagglutination-inhibiting (HI) antibody with one laboratory acting as a reference laboratory to provide a uniform basis for comparison of the HI results. The international unitage equivalent to the MIT used by the ten laboratories was found to vary from 24 to 98 units. Testing of the sera by immunofluorescence and by HI after flotation centrifugation indicated that residual non-specific inhibitors may interfere with HI antibody testing to an extent equivalent to 12-15 units. An acceptable MIT would therefore be equivalent to 24-48 units of rubella HI antibody. The single radial haemolysis (SRH) results on the sera indicate that this is a sensitive and specific test for rubella antibody.

Antibodies, Viral↗