Biomedical subjects

R E Randall

Publications and source records attributed to R E Randall.

At least 73 records · Page 4Linked to original sources

PubMed1984

A single major immediate-early virus gene product is synthesized in cells productively infected with herpesvirus saimiri.

A major virus-specific immediate-early (IE) polypeptide with an apparent mol. wt. of 52 000 (52K) was synthesized immediately after the removal of a cycloheximide block applied to owl monkey kidney cell cultures at the time of infection with high multiplicities of herpesvirus saimiri (HVS). The IE 52K polypeptide was phosphorylated and accumulated rapidly and efficiently in the nucleus of infected cells. Monoclonal antibodies to IE 52K and to a major non-structural delayed-early (DE) DNA-binding protein with similar electrophoretic mobility (DE 51K) were used to show that the IE 52K protein and DE 51K proteins are antigenically distinct. The monoclonal antibody to the IE 52K protein of HVS strain 11 reacted in immune precipitation and immunofluorescence tests with polypeptides of similar mol. wt. present as nuclear antigens in cells infected with heterologous strains of HVS and herpesvirus ateles.

Animals↗

PubMed1984

Isolation and characterization of monoclonal antibodies to structural and nonstructural herpesvirus saimiri proteins.

An improved screening procedure was applied to identify hybridomas secreting antibodies to herpesvirus saimiri-specified polypeptides among the products of fusions between SP2/0 myeloma cells and spleen cells from mice immunized with purified virus particles or virus-specific DNA-binding proteins. Twenty-four monoclonal antibodies were isolated with specificities for 13 different virus-specified polypeptides (or complexes of polypeptides), including the major capsid protein of the virus (150K), the 160K and 130K structural proteins, a 108K structural phosphoprotein, structural glycoproteins, the nonstructural early 76K protein, early nonstructural DNA-binding proteins of 48 to 51K and 110K and the major immediate-early protein of 52K. Antibody to the virus 76K protein precipitated a host protein of 62K, and a number of antibodies specific for host proteins were also isolated. Antibody to the 52K immediate-early polypeptide precipitated the delayed-early 76K protein, whereas the antibody to the 76K protein did not precipitate the 52K polypeptide. These observations suggest the presence of epitopes common to virus and host proteins and an antigenic site common to an immediate-early and a delayed-early virus protein. The antibodies were used to examine the sites of intracellular accumulation of virus polypeptides, the formation of complexes of structural proteins, and the postsynthetic processing of virus proteins. The present collection of monoclonal antibodies provides a set of reagents with specificities for members of each of the major kinetically or functionally distinct classes of virus gene products.

Animals↗

PubMed1983

Preparation and uses of immunoabsorbent monolayers in the purification of virus proteins and separation of cells on the basis of their cell surface antigens.

A method is described for forming monolayers of Staphylococcus aureus Cowan strain A on plastic tissue culture plates (A plates). Bacteria remain bound to such plates even under strong protein denaturing conditions. The specific binding of antibody to A plates provides a rapid method for immune precipitation on a solid matrix. Antibody can be covalently crosslinked to staphylococcus monolayers by fixation with paraformaldehyde without significant loss of specific antigen binding capacity. Furthermore antigen-antibody complexes may be efficiently disrupted with 9 M urea and non-ionic detergent and both antibody and antigen renatured by removal of urea, antibody regaining the ability to bind fresh antigen. In the presence of 9 M urea and 1% Nonidet P-40 fixed antibody remains bound to plates while antigen is released into the urea solution, providing a method for the immunological purification of proteins. Plates with such fixed antibody may be used multiple times to bind antigen. The use of this method is illustrated by the purification of 2 adenovirus structural proteins and a herpes simplex virus glycoprotein, by means of specific monoclonal antibodies to these proteins crosslinked to A plates. A method is also described to enrich for functional antibody from immune precipitates by dissociating antibody-antigen complexes with 9 M urea and 1% Nonidet P-40 and isolating the antigen free antibody on Staphylococcus aureus Cowan strain A. A plates have also been used to separate suspension cells on the basis of their cell surface antigens (e.g., lymphocyte subpopulations or cells expressing virus antigens on their surface). Cells were either (1) reacted with specific antisera to cell surface determinants and panned over A plates, cells with antibody on their surface binding to the monolayer, or (2) panned over A plates to which specific antibody had been coupled. Separated cells may be probed directly for certain properties, such as their ability to support virus replication. The staphylococcus monolayer does not prevent the growth of tissue culture cells on the plastic surface and the selected cell population can be examined for cells giving rise to infectious centres by the subsequent addition of permissive cells.

Animals↗

PubMed1983

Proteins specified by herpesvirus saimiri: identification and properties of virus-specific polypeptides in productively infected cells.

The synthesis and accumulation of more than 30 virus-induced polypeptides was detected after infection of permissive primate cells with high multiplicities of herpesvirus saimiri. These virus-induced polypeptides had apparent mol. wt. of from 12,000 (12K) to 250K and differed in molar abundance by up to two orders of magnitude. The majority of virus-induced polypeptides satisfied multiple criteria for their virus specificity. Polypeptides of 110K, 76K, 51K and 31K to 29K were synthesized at maximum rates early in infection, but the majority of proteins were made at high rates late in infection. Virus-specific polypeptides were substrates for a number of post-synthetic modifications. The 117K, 85K, 76K, 31K and 30K polypeptides were each processed to forms with altered electrophoretic mobility. The 117K and 85K polypeptides were among the virus-specified substrates for glycosylation and virus-induced polypeptides of 59K, 51K, 30K and 26K were phosphorylated. The early 51K phosphoprotein and the 30K to 31K polypeptides were rapidly translocated to the nucleus of infected cells. The 31K polypeptide was processed to a 29K product which remained stably associated with the nuclear fraction. The 30K nuclear protein was shown to be the precursor of a 28K polypeptide which was released in a soluble form into the cytoplasmic fraction and the culture medium of cells at late times in the virus growth cycle. Many other polypeptides accumulated slowly in nuclear (e.g. 250K, 150K, 130K, 110K and 38K) or in cytoplasmic (e.g. 117K, 85K and 28K) fractions of infected cells in forms which could be differentiated by the use of detergents or differences in stability to salt extraction. The mol. wt., relative molarities and some features of the post-synthetic processing of herpesvirus saimiri polypeptides more closely resembled the properties of gene products of Epstein-Barr virus than those of herpes simplex virus.

Animals↗

PubMed1982

Proteins specified by Herpesvirus saimiri: purification and properties of a single polypeptide which elicits virus-neutralizing antibody.

A virus-specified polypeptide of 160,000 mol. wt. (160K) was purified more than 1000-fold from the soluble proteins present in the culture medium of cells productively infected with herpesvirus saimiri (HVS). Purified preparations of the 160K polypeptide gave rise to high titre precipitating and neutralizing antibodies in immunized rabbits, which cross-neutralized independent isolates of HVS (neutralization rate constants, k, of between 3.8 and 4.8) and specifically precipitated the 160K polypeptide from extracts of cells infected with the homologous or heterologous strains of this virus. Antiserum to the 160K polypeptide of HVS also gave low (k = 0.02) levels of neutralization of the related herpesvirus ateles. Purified preparations of the 160K polypeptide were capable of removing most or all of the neutralizing antibody sera of squirrel monkeys with naturally acquired antibodies to HVS. The 160K polypeptide was previously shown to form part of the surface of enveloped virus particles. However, we show here that the 160K polypeptide is not extensively glycosylated in infected cells. The majority of glucosamine incorporation specific to HVS-infected cells was into eight regions with apparent mol. wt. of 170K to 220K, 125K to 145K, 115K to 120K, 83K to 88K, 65K to 75K, 52K to 58K, 25K to 27K and 12.5K to 13K. It remains possible that alternative cleaved or glycosylated forms of the 160K polypeptide are also present on the virus particle or that precipitating antibody to 160K and virus-neutralizing antibodies are not identical. However, our inability to detect precipitating antibody to virus-induced proteins or glycoproteins other than the 160K polypeptide and the high titre of neutralizing antibody present in this serum, provides reasonable evidence that it is the antibodies which react with the 160K polypeptide that are responsible for virus neutralization.

Animals↗

PubMed1980

Glycoproteins with type common and type specific antigenic sites excreted from cells infected with herpes simplex virus types 1 and 2.

Agar immunodiffusion tests demonstrated that BHK 21 cells infected with either HSV I or HSV 2 release only a few HSV-specified antigens into the extracellular fluid (infected cell released polypeptides-ICRP). Neutralization blocking experiments showed that the majority of antigens/(including the Band II common antigen) involved as target sites in antibody-mediated virus neutralization are present in the ICRP of both HSV I and HSV 2.SDS-PAGE identified six regions of virus-specified proteins in the ICRP from both HSV I- and HSV 2-infected BHK cells. All these specifically released proteins are glycosylated, although to varying degrees. The SDS-PAGE profiles of HSV I and HSV 2 ICRP are different but do show some similarities, the most notable being a highly glycosylated protein with an estimated mol. wt. of 50,000 to 54,000 in HSV I ICRP and 52,000 to 56,000 in HSV 2 ICRP. Immune precipitation demonstrated that these two proteins contain the Band II antigenic site. Similar studies showed that the major type I specific antigenic site, which is involved as a target site in the neutralization of virus infectivity, is located in the highest mol. wt. glycoprotein region of HSV I ICRP and has a similar mobility to the VP7/8 region of purified enveloped virus.

Animals↗

PubMed1980

Proteins of herpesvirus saimiri: identification of two virus polypeptides released into the culture medium of productively infected cells.

Two virus-induced polypeptides are preferentially released into the culture medium of owl monkey kidney cells productively infected with herpesvirus saimiri (HVS). These polypeptides have apparent mol. wt. of 160 X 10(3) and 28 X 10(3) and are the major virus-induced polypeptides in a membrane fraction from infected cells. Both these excreted polypeptides co-migrate with virus structural proteins and are selectively released from enveloped virus particles by treatment with Nonidet-P40.

Animals↗

PubMed1978

Observations of antigenic relatedness between viruses of the herpes simplex "neutroseron".

The antigenic relatedness of three viruses of the herpes simplex type 1 neutroseron - herpes simplex virus types 1 (HSV-1) and 2 (HSV-2) and bovine mammillitis virus (BMV) - has been examined by immune precipitation and virus neutralization tests. Many virus-specific infected-cell polypeptides were shown to possess antigenic sites shared by both HSV-1 and HSV-2. Cross-neutralization between the viruses is mediated through antibodies to at least two antigenic sites, one shared by HSV-1, HSV-2 and BMV and one shared by HSV-1 and HSV-2 but not BMV.

Antigens, Viral↗

PubMed1977

Some properties of recombinants between type 1 and type 2 herpes simplex viruses.

Four intertypic recombinants of herpes simplex virus have been shown to possess genetic information for functions characteristic of each of the two parental types. The functions were identified by (a) polyacrylamide gel electrophoresis of purified virus particles and of polypeptides synthesized in cells infected with the recombinants and (b) analysis of antigenic sites interacting with type specific neutralizing antibody. The analysis shows that each recombinant possess a different combination of these type specific markers. Finally we have been unable to detect recombination between herpes simplex type 1 and pseudorabies viruses.

Animals↗

PubMed1976

Manifestations of systemic light chain deposition.

In two patients with terminal renal failure, manifestations of disease developed in multiple organ systems. One had a previous diagnosis of multiple myeloma with kappa light chain proteinemia and proteinuria. The other had idiopathic lobular glomerulonephritis. Hepatic and neurologic abnormalities developed in both; in the latter gastrointestinal, cardiac and endocrine disease developed as well. Clinical and pathologic correlations suggest that the retention and tissue deposition of light chains produced the organ dysfunction, inasmuch as free kappa light chain determinants were demonstrated histologically in the clinically affected organs. The deposition in these patients may be an extreme example of a common but previously unrecognized form of plasma cell dyscrasia.

Adult↗

PubMed1975