Biomedical subjects
Publications and source records attributed to D D Porter.
A second nonstructural protein of the Aleutian disease parvovirus was predicted from nucleotide sequence analysis and a detailed transcription map. Western immunoblotting analysis showed that infected mink and ferrets show an antibody response to this predicted protein.
Adenoviruses can produce severe disease, especially in patients who are immunosuppressed. We present a unique case in which adenovirus type 5 was demonstrated retrospectively, by using immunohistochemical methods, in the appendix of a liver transplant donor. The donor had intussusception, which has been associated with adenovirus infection. Both the initial and a second liver transplant in the recipient were severely damaged by adenovirus type 5. These findings were demonstrated immunohistochemically, as well as on electron microscopy. The recipient died due to the infection and ensuing complications.
Although herpes simplex virus type 1 (HSV-1) is known to reside latently in trigeminal ganglia between episodes of reactivation, the mechanisms involved in restricting the virus to this state are not understood. Using in situ nucleic acid hybridization methods, we show that there is HSV-encoded RNA in ganglion cells from 10 of 12 seropositive and zero of three seronegative patients studied at autopsy. Transcripts mapping to the region encoding the immediate-early polypeptide ICPO and the latency-associated transcript (LAT) were detected in the nuclei of these neurons. No other region of the HSV-1 genome was found to be expressed. When carefully defined probes were used to identify the transcripts, RNA corresponding to the LAT and, rarely, to ICPO was found. These results suggest that HSV-1 latency is an active process and that the LAT may be involved in regulating viral genetic expression.
Respiratory syncytial virus (RSV) is a cause of significant morbidity and mortality in infants and children with an immunocompromised status or a congenital heart disease. The following case describes a 6 8/12-year-old, previously normal child who had a fatal interstitial pneumonitis caused by RSV. Documentation of RSV as the etiologic agent and documentation of her immune status are presented. In light of recent advances in the rapid diagnosis and treatment of RSV, this virus should be considered in children with an unusual interstitial pneumonitis regardless of their known immunologic status.
The majority of ferrets infected with a ferret strain of Aleutian disease virus (ADV) produce antibody only to a detergent-sensitive common determinant on the two closely related virion proteins. Ferrets with high antibody titers and mink infected with this virus also produce antibody to one or more virion immunogenic determinants unaffected by detergent.
The mechanism of viral clearance was studied by using the mouse model of chronic infection with lymphocytic choriomeningitis virus. Distinct patterns of viral clearance and histopathology were observed in different organs after adoptive immune therapy of persistently infected (carrier) mice. Clearance from the liver occurred within 30 days and was accompanied by extensive mononuclear cell infiltrates and necrosis of hepatocytes. Infectious virus and viral antigen were eliminated concurrently. This pattern of viral clearance was also seen in most other tissues (i.e., lung, spleen, lymph nodes, pancreas, etc.). In contrast, a different pattern of clearance was observed in the brain. Infectious virus was eliminated within 30 days, but viral antigen persisted in the central nervous systems of treated carrier mice for up to 90 days. The urinary system was the most resistant to immune therapy. Elimination of infectious virus and viral antigen from the kidney took greater than 200 days and even then was not complete; trace levels of infectious virus were still present in the kidneys of some treated carrier mice. After immune therapy, viral antigen in the kidney was located within renal tubules that costained for intracellular mouse immunoglobulin G. This unusual staining pattern, coupled with the observation of large numbers of plasma cells within the kidney, suggests that virus-immunoglobulin G complexes found in the tubules may represent in situ immune complex formation as opposed to deposition of circulating immune complexes. In conclusion, these results suggest that the site (organ) of viral persistence is an important consideration in developing treatment strategies for controlling chronic viral infections.
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Separation of mixtures of proteins by polyacrylamide gel electrophoresis followed by transfer of the proteins to support media such as nitrocellulose and detection by immunologic procedures provides a powerful analytic tool for assaying the antibody specificity of antisera or for following the purification of antigens. This technique requires fewer assumptions about antigenic solubility or antibody reactivity than immunoprecipitation methods. We present a glucose oxidase immunoenzyme staining procedure for protein blots and illustrate its use for detecting antibody to several viruses. The glucose oxidase immunoenzyme stain has a lower background than some peroxidase stains. We have detected as little as 1 microgram/ml of antiviral antibody using this stain.
Aleutian disease virus (ADV) persistently infects mink and causes marked hypergammaglobulinemia. Immunoglobulin class-specific antisera were used to define the total immunoglobulin of each class by radial immunodiffusion and the immunoglobulin class of ADV-specific antibody by immunofluorescence in experimentally and naturally infected mink. Electrophoretic gamma globulin closely reflects the immunoglobulin G (IgG) level in mink, and the majority of the increased immunoglobulin and ADV antibody in infected mink is IgG. IgM becomes elevated within 6 days after infection, reaches peak levels by 15 to 18 days, and returns to normal by 60 days after infection. The first ADV antibody demonstrable is IgM, and most mink have virus-specific IgM antibody for at least 85 days postinfection. Serum IgA levels in normal mink are not normally distributed, and ADV infection causes a marked elevation of IgA. Low levels of ADV-specific IgA antibody can be shown throughout the course of infection. Failure of large amounts of virus-specific IgG antibody to inhibit the reaction of virus-specific IgM and IgA antibodies suggests that the various classes of antibodies are directed against spatially different antigenic determinants. The IgM and IgA were shown not to be rheumatoid factors.
Aleutian disease virus (ADV), an autonomous parvovirus, persistently infects mink and induces very high levels of virus-specific antibody. All strains of ADV infect all mink, but only highly virulent strains cause progressive disease in non-Aleutian mink. The development of antibody to individual ADV proteins was evaluated by Western blotting by using the sera of 22 uninfected mink and 163 naturally or experimentally infected mink. ADV has virion proteins of 86,000 and 78,000 daltons that are closely related. A new, possibly nonvirion protein of 143,000 daltons was observed, as well as a known nonvirion protein of 71,000 daltons. Sera from mink experimentally or naturally infected with ADV of high or low virulence generally reacted about equally with all four proteins. The only exceptions noted were that 8 of 15 sera of mink infected transplacentally preferentially reacted with the two virion proteins and sera from mink with the monoclonal gammopathy of Aleutian disease reacted preferentially with either virion (10 of 12) or nonvirion (2 of 12) proteins.
Aleutian disease (AD) is caused by a persistent infection of mink with an autonomous parvovirus. Chronically infected mink develop widespread plasmacytosis, a marked elevation of their serum IgG, and immune complex disease. A substantial fraction of the IgG in the serum of mink with Aleutian disease may be specifically absorbed by monolayer cell cultures infected with Aleutian disease virus. The maximum percentage of absorption of IgG found was 81% in a mink with 5.4 g/dl of IgG. Mink with the monoclonal gammopathy of Aleutian disease had a particularly large percentage of the IgG absorbed. The percentage of IgG absorbed from serums of mink with Aleutian disease is directly proportional to the serum IgG level and to the Aleutian disease viral antibody titer. The amount of IgG which can be absorbed by infected cell monolayers increases during the course of experimental infection, and the absorption is immunologically specific. Thus, it appears that much of the hypergammaglobulinemia in mink with Aleutian disease represents virus-specific antibody.
Autopsy slides from 22 melanoma patients who received bacille Calmette-Guérin BCG immunotherapy and who had a postmortem examination at the UCLA Center for the Health Sciences were examined for the presence of granulomas. Granulomas were found in six patients (29%) who received BCG but not in a group of 25 melanoma patients who did not have BCG immunotherapy. A number of factors were tested for correlation with the presence of granulomas in the autopsy material. Fifty-five percent of the patients who received BCG by both intralesional and time technique developed granulomas. No patients given BCG by the tine technique alone developed granulomas. No correlation was found between granulomas and the presence of symptoms after the administration of BCG, the duration of BCG immunotherapy, the patient's age, the number of BCG administrations, treatment with immunosuppressive agents, the length of survival after the last BCG administration, the presence of a positive PPD reaction, a positive history for granuloma-forming diseases and tests for immunocompetence. However, these results provide evidence that the route of BCG administration strongly influences the frequency of granulomas in melanoma patients who received BCG immunotherapy.
When 32 antibody-free ferrets were inoculated with the highly mink-virulent Utah-1 strain of Aleutian disease virus (ADV), most developed ADV antibody starting 15 days after infection, but the antibody titers were much lower than those seen in mink. Relatively small amounts of ADV were demonstrated in CRFK cell culture, using ferret spleen and lymph node homogenates only 4 to 10 days after experimental infection, but low-level viral persistence for 180 days was shown by mink inoculation. The ferrets inoculated with the Utah-1 strain of ADV did not develop elevated gamma globulin levels, but did have mild tissue lesions. Forty-two percent of a group of 214, approximately 1-year-old, recently pregnant, female ferrets were found to have antibody to ADV. An analysis of the serum proteins of the ferrets with ADV antibody showed that they had a significant, but mild, elevation of their serum gamma globulin. Serial ferret-to-ferret transmission of a ferret strain of ADV by inoculation of spleen homogenates was demonstrated, and some of these ferrets developed liver lesions. Mink inoculated with ferret ADV made antibody, but did not develop hypergammaglobulinemia or tissue lesions. Although both ferret and mink strains of ADV replicate and persist in the ferret, they fail to cause severe disease of the type usually seen in the closely related mink. Mink and ferret ADV strains appear to be biologically distinct.
A term newborn suffered disseminated herpes simplex virus (HSV) type II infection five days after cesarean section delivery for fetal distress. The mother had no history or evidence of herpetic lesions; the father had a history of genital herpetic lesions. The infant's terminal course was dominated by disseminated intravascular coagulation (DIC) with hepatic and renal failure. Microscopic examination revealed a necrotizing vasculitis of small and medium-sized lung and peripancreatic arteries. Nuclear inclusions characteristic of HSV were found in these arteries, as well as in the adrenal parenchyma, spleen, and lymph node; electron microscopy confirmed replication of virus within the arterial endothelial cells. The mechanism of arterial damage in severe herpetic infection contrasts with the immune-complex mechanism postulated for other viral vasculitides. Direct, virally induced arterial damage resulting in exposure of collagen may set the stage of DiC, a commonly fatal complication of this disease.
Enzymes as markers for antigens or antibodies in immunohistochemical procedures have several advantages over commonly used fluorochrome labels. These include use of a regular light microscope and the ability to get permanently stable slide preparations. Glucose oxidase (EC 1.1.2.3.4), being absent in mammalian tissue, provides no background staining, such as that seen with the commonly used horseradish peroxidase (EC 1.11.1.7) owing to peroxidase-like activity in tissues. A glucose oxidase histochemical method is detailed that is useful for detection of human antibodies; it can be easily used in clinical laboratories as a substitute for fluorescent techniques.
Illness associated adenovirus infection is described in 15 immunocompromised patients. Patients were immunocompromised by severe underlying disease, immunosuppressive or corticosteroid therapy or by age (prematurity). Evidence of adenovirus infection was obtained by either viral isolation or, in two cases, characteristic adenovirus inclusion bodies at postmortem study. All clinical illness was associated with high fever (temperature greater than 39 degrees C). Eighty per cent of the patients had severe systemic complaints including malaise, lethargy, fatigue and night sweats; a similar number of gastrointestinal symptoms. Pulmonary complaints were described in 11 of 15 cases and included cough (67 per cent) and tachypnea (53 per cent). Roentgenologic evidence of pneumonia was demonstrated in 12 of 15 patients (80 per cent). Elevation of serum hepatic enzyme levels (serum glutamic pyruvic transaminase (SGPT)) occurred in eight of 11 patients (73 per cent) and was moderate to severe (serum glutamic pyruvic transaminase greater than 450 IU/liter) in five of 11 (45 per cent). Nine patients died; seven after a rapid downhill course and two after a prolonged illness. Evidence of adenovirus infection microscopically by autopsy in the lung, liver or both is demonstrated in four patients with fulminant systemic illness. Adenovirus infection should be considered in the etiology of severe overwhelming illness in the immunocompromised host.
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