David L. Madden

Evaluation of enzyme-linked immunosorbent assay (ELISA) for mumps virus antibodies.

Summary Pre- and postvaccination sera from individuals who had been vaccinated with live Jeryl-Lynn vaccine were assayed for mumps antibodies by using serum neutralization (NT), enzymed-linked immunosorbent assay (ELISA), hemolysis-in gel (HIG), hemagglutination inhibition (HI) and complement fixation (CF) techniques. ELISA was found to be equally specific and somewhat more sensitive than NT. HIG was less sensitive than either of these two and, with some specimens, less specific. However, it was more sensitive than HI. CF gave several low positive readings which were interpreted as false positive since they were found in prevaccination sera and other tests failed to detect antibodies in these samples. The ELISA mumps test can be used in place of the more expensive and time consuming neutralization test for screening epidemiology, documenting immunity and establishing the serological diagnosis.

Humoral and cell-mediated immune responses to herpesvirus antigens during pregnancy—A longitudinal study

Humoral and cellular immune responses were studied during the second trimester, third trimester, and postpartum periods in 11 pregnant women and in nonpregnant control women. Complement fixing (CF) and indirect hemagglutinating antibody (IHA) titers for herpes simplex type 1 (HSV-1), herpes simplex type 2 (HSV-2), and cytomegalovirus (CMV) were determined. Cellular response was measured by [3H]thymidine uptake by stimulated lymphocytes. Phytohemagglutinin (PHA), HSV-1, HSV-2, and CMV antigens were used as stimulants. No differences in the mean titers of CF and IHA antibodies were found. The cellular response to PHA had a transient decrease (P<0.02) during the third trimester. The cellular response to CMV was significantly lower during the second and third trimesters. A diminished response to HSV-1 antigen was observed during the second and third trimesters; the cellular response to HSV-2, though reduced, was not significantly altered during pregnancy. These data indicate a suppression of cellular responses to various herpesviruses and PHA during pregnancy.

Immune defects in simian acquired immunodeficiency syndrome

We recently reported a Simian Acquired Immunodeficiency Syndrome (SAIDS) in rhesus macaques at the California Primate Research Center. Here, we studied in vitro lymphocyte response to the mitogens Concanavalin A (Con A), phytohemagglutinin (PHA) and pokeweed mitogen (PWM) with and without interleukin 2 (IL-2). Immunoglobulin (IgG and IgM) and complement (C3 and C4) concentrations were determined by radial immunodiffusion. T helper and T suppressor lymphocytes were identified with the monoclonal antibodies OKT4 and OKT8. Concentrations of IgG and IgM were significantly (p less than .05) decreased. Complement component C3 did not change but C4 was increased. The absolute lymphocyte count decreased but the OKT4:OKT8 ratio was unchanged from controls. A decreased lymphocyte response to all mitogens occurred early and became more severely depressed near death. IL-2 caused a complete or partial restoration of the response to the mitogens CON A and PHA. Both the humoral and cell mediated immune responses are affected in SAIDS. The role of IL-2 in this immune defect must be studied further.

The occurrence of epidemic infectious hepatitis in chronic carriers of Australia antigen.

During a major institutional epidemic of infectious hepatitis, chronic carriers of Australia antigen were identified and found to be susceptible to infectious hepatitis. This observation provides additional evidence that serum hepatitis, which has been associated with Australia antigen, and infectious hepatitis, which is not associated with the antigen, are immunologically dissimilar and caused by different etiologic agents. The institutional frequency and pattern of Australia antigenemia is reported. The antigen was present in 23 per cent of patients with Downs syndrome and in 6 per cent of other mentally retarded patients. The antigen was more prevalent in younger patients than in older patients and twice as prevalent in males as in females.

Simian varicella virus (delta herpesvirus) infection of patas monkeys leading to pneumonia and encephalitis.

Abstract Patas monkeys were infected with Delta herpesvirus (DHV) to determine the pathogenicity of this virus in the lung and brain in order to assess the suitability of this model for severe complications in human VZV infection. Clinically, pneumonia and encephalitis occurred following intracerebral inoculation. Intratracheal or intravascular inoculation of DHV produced pneumonia but not encephalitis. Prominent microscopic lesions included diffuse interstitial pneumonia and focal hemorrhagic encephalitis. Typical herpesvirus intranuclear inclusion bodies were noted in lung parenchymal cells and in glial cells of the brain. DHV was isolated from peripheral blood leukocytes, spleen, lymph nodes, lung, and from various sites in the peripheral and central nervous system. Fluorescent antibody staining confirmed the identity of DHV recovered from monkey tissue. This study provides further evidence that the patas-DHV model has potential for predicting drug and/or immunotherapy regimens which may be useful in treating pulmonary and neurological complications of VZV infection.

Antibody to human and simian retrovirus, HTLV-I, HTLV-II, HIV, STLV-III, and SRV-I not increased in patients with multiple sclerosis.

We have tested sera from patients with multiple sclerosis, matched controls, and those with other neurological diseases, as well as sera from patients with the acquired immunodeficiency syndrome and controls and patients with tropical spastic paraparesis (TSP) and controls for antibody to human T‐lymphotropic virus type I (HTLV‐I), HTLV‐II, human immunodeficiency virus (HIV), simian T‐lymphotropic virus type III, or simian retrovirus type I by immunofluorescent activity test, and for HTLV‐I and HIV by the ELISA method. Sera from patients with multiple sclerosis and matched controls, and from patients with optic neuritis and Parkinsons or other neuromuscular diseases did not have antibody to any of the retroviruses tested. Specimens from TSP patients and some controls contained HTLV‐I antibody. We conclude from our study that only TSP patients had serological evidence of infection with one of the retroviruses studied.

Measles antibody titers in multiple sclerosis patients and HLA‐matched and unmatched siblings

Antibody titers to measles, cytomegalovirus, and herpesviruses 1 and 2 were compared for siblings of multiple sclerosis (MS) patients sharing two, one, and no histocompatibility antigen haplotypes with the case. Significant differences were observed only for measles. Titers were significantly lower in siblings sharing no haplotypes with the case. Within case-sibling pairs, the presence of HLA-A3 and/or B7 affected measles antibody titers more than the presence of MS. These findings suggest that the immune response to measles in these sibships is influenced by the presence of HLA-A3 and/or B7 as well as another familial factor.

Comparison of sensitivity of radioimmunoassay and immune electron microscopy for detecting hepatitis A antigen in fecal extracts.

Summary Solid-phase, microtiter ra-dioimmunoassay (RIA) has been compared with immune electron microscopy (IEM) for its sensitivity and usefulness in detecting hepatitis A antigen in clinical fecal specimens. Using relatively crude 5% extracts from stools collected at the Lynchburg Training School and Hospital in 1970 during an epidemic of infectious hepatitis, we have demonstrated the feasibility of RIA as a rapid and reliable screening test. All samples positive for HA Ag by IEM were also positive by RIA. Dilution experiments with HA Ag-positive fecal preparations indicated that RIA is at least as sensitive as IEM. RIA is an appropriate method for reliable, large-scale screening of fecal samples for HA Ag. RIA would appear to be potentially useful in the diagnosis of hepatitis A infection.

Virus antibodies in the cerebrospinal fluid of multiple sclerosis patients detected with ELISA tests

Abstract The enzyme-linked immunosorbent assay (ELISA) was used to determine levels of specific IgG antibodies against measles, rubella, vaccinia, corona (OC43) and mumps viruses in cerebrospinal fluid (CSF) and serum of 18 patients with clinically definite multiple sclerosis (MS), 8 patients with optic neuritis (ON), 27 patients with other neurological disease (OND), and 88 control subjects without central nervous system disease. Serum antibody levels were not significantly different between the four groups. Differences in the frequency and levels of CSF antibodies between the four groups were observed. Control patients had serum CSF antibody ratios from 2.0 to 3.0 (log) with an average of 2.5 corresponding to a 320-fold difference between serum and CSF antibody levels. MS patients had ratios from 1.1 to 2.1 with an average of 1.6. The average was 2.0 for the ON patients. The average for the OND patients was similar to the controls. The altered serum CSF ratios for several viruses within an individual patient was similar. These results suggest that nonspecific immunostimulation is responsible for the increased levels of CSF virus antibodies.

The Etiology of Non-Specific Urethritis in Active Duty Marines

We studied patients with non-specific urethritis and control subjects at our dispensary. These patients and controls were matched for age, rank and sexual activity, and studied for the presence of bacteria, virus, Trichomonas and Mycoplasma. No significant bacteria were found in either group. No Trichomonas was identified, only 1 herpes II was recovered and no cytomegalovirus was found. Mycoplasma were cultured from patients and controls. The rate of colonization varied, depending upon several factors, but the significant factors seemed to be associated with the number of sexual partners. Those men with more than 1 sexual partner had a significantly increased colonization with Mycoplasma.