Barry Ziola

Solid-phase radioimmunoassay of herpes simplex virus IgG and IgM antibodies.

A solid-phase radioimmunoassay for detection of herpes simplex virus-specific IgG and IgM antibodies in human serum specimens is presented. Virus antigen is adsorbed on polystyrene balls and antibodies which attach to the antigen are detected by 125I-labeled antihuman-gamma or antihuman-mu immunoglobulins. A total of 76 specimens have been tested. The appearance of virus-specific IgG and IgM antibodies in primary herpetic infections was readily demonstrated. When serum samples from patients with past exposure to herpes simplex virus were tested, endpoint titers of virus-specific IgG antibodies were found to be 8 to 2048 times higher than titers determined by a complement fixation test. Apparent cross reactivity with varicella-zoster virus was observed in the present radioimmunoassay.

Polystyrene balls as the solid-phase of a double-antibody radioimmunoassay for human serum albumin

Polystyrene balls have been incorporated as the solid-phase of a model double-antibody radioimmunoassay for human serum albumin. Purified IgG from the secondary antiserum is adsorbed on the 6.4 mm diameter balls. The solid-phase secondary antibody is then used to separate primary antibody bound iodinated antigen from unbound antigen. The secondary antibody coated polystyrene balls are easily prepared and manipulated; several hundred sample dilutions can readily be processed in a single assay. Assay background values of 1.5% or less are consistently obtained without extensive or special washing procedures.

Stimulation of lymphocytes from subacute sclerosing panencephalitis patients by defined measles virus antigens

Abstract The lymphoproliferative response of human peripheral blood mononuclear cells to different measles virus antigen preparations was studied with lymphocytes from 38 measlesseropositive healthy donors and 4 subacute sclerosing panencephalitis patients. The response was very weak or absent in all of the controls and in three of the subacute sclerosing panencephalitis patients. The fourth subacute sclerosing panencephalitis patient had fluctuating levels of lymphocyte stimulation by measles antigens. The response was very strong for several months and during this time the parameters of the test system were characterized. It was discovered that a membrane preparation of measles-infected cells caused stimulation equal to that of highly purified virions. Purified measles ribonucleoprotein also induced specific stimulation, although lower than that seen with other types of measles antigens. Results of experiments on stimulation kinetics and antigen dose responses were compatible with antigen-specific stimulation. Enriched T cells were more vigorously stimulated than unfractionated peripheral blood mononuclear cells suggesting that this transformation test is specific for T cells.

OC43 strain-related coronavirus antibodies in different age groups

Serum antibodies against human coronavirus OC43 in different age groups were measured by complement fixation (CF), haemagglutination inhibition (HI), radial diffusion haemolysis‐in‐gel (HIG), and solid‐phase radioimmunoassay (RIA) methods. Antigen grown in suckling mouse brain was used in all tests. Results obtained by the CF and HIG tests, and the RIA, were in good agreement with regard to the presence or absence of antibodies. Similar results were also obtained with the HI test if nonspecific haemagglutination inhibitors were first removed by treatment with phospholipase C and only titers of 1:20 or greater were considered positive.

Measles virus-specific IgM antibodies and IgM-class rheumatoid factor in serum and cerebrospinal fluid of subacute sclerosing panencephalitis patients

Abstract Serum and cerebrospinal fluid (CSF) from 20 subacute sclerosing panencephalitis (SSPE) patients were analyzed with solid-phase radioimmunoassays designed to detect measles virus-specific IgM antibodies and IgM-class rheumatoid factor (RF). Serum and CSF from patients 1 to 16 did not contain measles IgM antibodies or elevated levels of RF. Serum from all and CSF from two of the remaining four patients were apparently measles IgM positive. RF levels were elevated, however, in serum from two and in serum and CSF from another of these four patients. Testing after removal of IgG or RF revealed that patient 17 had both measles IgM and elevated levels of RF in serum, and measles IgM in CSF. Reactivation of measles IgM synthesis was detected in the blood of patient 18. The serum of patient 19 and the serum and CSF of patient 20 contained only elevated levels of RF. The implications of these results are twofold. First, although measles IgM is found in some SSPE patients, it is not found consistently enough to be a marker for the chronic measles virus infection in these patients. Second, the possible interfering presence of RF must be taken carefully into account in any study of IgM antibody persistence.

Radioimmunoassay of herpes-simplex and measles virus antibodies in serum and cerebrospinal fluid of patients without infectious or demyelinating diseases of the central nervous system.

A solid-phase radioimmunoassay was used to detect IgG antibodies against herpes-simplex virus antigens (capsid, envelope and excreted) and against measles virus antigen in serum and cerebrospinal fluid (CSF) specimens of 61 patients with no evidence of infectious or demyelinating disease of the central nervous system. Quantitative determinations of IgG and albumin in serum and CSF were also performed. Of the 61 serum and 61 CSF samples tested, 57 and 56 respectively contained antibodies against subunit antigens of herpes simplex virus. Antibody against measles virus was found in 59 serum and 47 CSF specimens. A positive correlation (P less than 0-001) was found between each of the four serum to CSF antibody ratios and the serum to CSF total IgG ratios. This indicated that the distribution of antiviral IgG antibodies in serum and CSF normally follows the distribution of total IgG. The ratios between viral antibody in serum and CSF were also correlated with albumin ratios (P less than 0-05). An inverse relation (P less than 0-001) was found between the age of the patients and their serum to CSF albumin ratios, but not their IgG ratios, suggesting that the albumin ratio is a useful indicator of a blood brain barrier lesion and that the IgG ratio should be used in evaluating disturbed antibody ratios.

Coincidental fluctuations of humoral immunity and clinical progression in a patient with subacute sclerosing panencephalitis

SummarySerum and cerebrospinal fluid were obtained from a 6-year-old male with subacute sclerosing panencephalitis (SSPE). Specimens were collected over a 9-month period beginning in the unusually acute phase and ending in a more quiescent phase of the disease. Immune complexes, auto-antibodies and viral antibodies were measured by radio-immunoassays. Fluctuations in these humoral immune parameters coincided with cessation of the acute phase of this disease. The results show that neurological changes in SSPE patients can be reflected in immune responses within both the peripheral circulation and the central nervous system.