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Biomedical subjects

G Miller

Publications and source records attributed to G Miller.

At least 415 records · Page 23Linked to original sources
PubMed1982

Neutralization of lymphocyte immortalization by different strains of Epstein-Barr virus with a murine monoclonal antibody.

A murine monoclonal antibody was raised against the B95-8 strain of Epstein-Barr virus (EBV), which was isolated from a case of mononucleosis after blood transfusion (Hoffman et al. Proc. Natl. Acad. Sci. U.S.A. 77:2979, 1980). We provide evidence that neutralization of immortalization by this monoclonal antibody is virus specific, since its potency was inversely related to the dose of challenge virus. Furthermore, the monoclonal antibody recognized antigens on viruses grown in human as well as in marmoset cells. We show that this monoclonal antibody neutralized three other transforming strains of EBV originating, respectively, from American patients with mononucleosis and fatal polyclonal lymphoma and from an African child with Burkitt lymphoma. However the antibody did not neutralize or detect antigens by immunofluorescence in the W91 strain of EBV. The hybridoma antibody did neutralize other EBV strains derived from the same Burkitt lymphoma cell line (Nyevu), as was the case with the W91 strain. This monoclonal antibody provides clear evidence of antigenic differences on the surface of EBVs and will ultimately prove useful in defining the antigenic site on EBV which elicits neutralizing antibody.

Animals↗
PubMed1982

Non-immortalizing P3J-HR-1 Epstein-Barr virus: a deletion mutant of its transforming parent, Jijoye.

The P3J-HR-1 strain of Epstein-Barr virus (EBV) fails to immortalize human lymphocytes. We wished to understand the nature of the genomic alterations which correlated with the loss of this ability. As a first step, the heterogeneity of DNA molecules in the P3J-HR-1 line was eliminated by cell cloning. Then a physical map was prepared of virion DNA from one cell clone, designated FF452-3. By comparison with the genomes of two EBVs, B95-8 and FF41, which are competent to immortalize lymphocytes, we identified a total of eight modifications of BamHI and EcoRI restriction endonuclease fragments of EBV (FF452-3) DNA consisting of insertions, deletions, or loss of a restriction endonuclease recognition site. To determine which of these alterations might be responsible for the loss of transforming phenotype, we examined homologous DNA fragments of the Jijoye strain of EBV, the progenitor of the HR-1 strain which still retains the ability to immortalize lymphocytes. We also studied viral DNA in lymphocytes transformed in vitro by Jijoye virus. Six of the eight alterations were found both in Jijoye and in clonal HR-1 DNA and were presumably genomic traits characteristic of this lineage of EBV. A small deletion in the BamHI-K fragment of HR-1 DNA was not found in Jijoye virion DNA, but this deletion was present in intracellular Jijoye DNA. Thus only one major genomic lesion in HR-1 DNA, a deletion of at least 2.4 x 10(6) molecular weight of DNA from a fused BamHI-H-Y fragment, consistently distinguished Jijoye DNA from its non-immortalizing P3J-HR-1 derivative. This deletion is likely to affect EBV genes which are directly or indirectly involved in immortalizing lymphocytes.

Base Sequence↗
PubMed1982

Immunoglobulin expression by human B lymphocytes clonally transformed by Epstein Barr virus.

Epstein Barr virus (EBV) immortalizes human B lymphocytes, resulting in long-term lymphoblastoid cell lines. To date, such cell lines have typically been derived by suspension culture of mixed mononuclear cells, infected in vivo or in vitro with EBV. We have derived 208 lymphoblastoid lines by clonal transformation of peripheral blood lymphocytes from 3 neonates, 3 normal adults, and 1 patient with systemic lupus erythematosus (SLE), utilizing colony formation in agarose and then separate propagation of the colonies in liquid medium. A total of 180 lines have been analyzed for expression of cytoplasmic and/or secreted immunoglobulin isotypes. Such analysis reveals that 167 (92%) of the lines are monoclonal with respect to isotype expression. All clonally derived lines from neonates produce only IgM. Most clonal transformants from normal adults also produce IgM (81%), but clones producing IgG (16%) or IgA (3%) were also found. Clonal transformation of lymphocytes from the SLE patient revealed a shift in isotype expression, with increased numbers of IgG-secreting clones (48%) and IgA-secreting clones (13%). The data support the use of EBV-mediated clonal transformation as a probe for the analysis of states of isotype differentiation among circulating B cells, and as a method for deriving immortalized, monoclonal human B cell lines.

Animals↗
PubMed1982

A monoclonal immunoblastic sarcoma in donor cells bearing Epstein-Barr virus genomes following allogeneic marrow grafting for acute lymphoblastic leukemia.

A patient undergoing marrow grafting for acute lymphoblastic leukemia from his partially HLA-mismatched sister displayed a widely disseminated immunoblastic sarcoma at autopsy. The tumor was monoclonal by immunoglobulin light-chain staining. Blot hybridization analysis, using a cloned highly polymorphic locus in human DNA as a probe, showed the tumor to be of donor-cell origin. Cytogenetic analysis also demonstrated donor-cell origin. Blot hybridization analysis demonstrated Epstein-Barr virus (EBV) genomes in the tumor. By contrast, reexamination of material from a previously reported case of a donor-type relapse showed no evidence of EBV DNA. In neither case was there evidence of cytomegalovirus DNA. This study documents the association of EBV with a malignant, monoclonal B-cell lymphoma arising in a marrow graft recipient. We conclude that DNA restriction fragment length polymorphisms can be used to prove the origin (donor or host) of neoplastic relapse following allogeneic marrow grafting. Further, cell types different from those of the original leukemia may be involved.

B-Lymphocytes↗
PubMed1982

Prune-belly syndrome.

A developmental defect of the abdominal wall musculature is commonly associated with urinary tract abnormalities and cryptorchidism. Rarely, a form of prune-belly syndrome occurs in females. A characteristic finding is the presence of dimpling on the outer aspect of the knees and elbows. In mild cases, no treatment is necessary; in the most severe cases, the infant is born without renal function.

Abdominal Muscles↗
PubMed1982

Gas chromatographic method for determining strychnine residues in alfalfa.

A gas-liquid chromatographic procedure is described for determining strychnine in alfalfa with a nitrogen-phosphorus detector. Alfalfa samples containing strychnine residues are chopped and extracted in a blender using ethyl acetate, sodium carbonate, and sodium sulfate, and the organic extract is extracted with 0.1N H2SO4 solution, made basic, and subsequently extracted with methylene chloride. Following concentration, the strychnine residues are quantitated by peak height, using an external standard. Recoveries for 0.05 ppm spikes were 88 +/- 11%.

Chromatography, Gas↗
PubMed1981

Retinoids and the prevention of cervical dysplasias.

Women with abnormal cytology were matched with normal control subjects for age, parity, ethnicity, and socioeconomic class and participated in a blind case-control study focused on the role of nutrition in cervical dysplasia. Sucrose gradient ultracentrifugation studies for determination of the presence and concentration of the binding proteins for retinol and retinoic acid were performed on colposcopic biopsy tissue specimens. The nutritional survey revealed statistically significant differences for vitamins A and C and beta carotene. Retinol binding protein was absent or minimally detectable and inversely related to the severity of the dysplasia. It is proposed that a double-blind clinical trial be conducted to evaluate whether retinoids may pharmacologically inhibit, arrest, or reverse cervical dysplasia.

Ascorbic Acid↗
PubMed1981

Epstein-Barr viral DNA: infectivity for human placental cells.

Purified DNA of Epstein-Barr virus (EBV) is regularly infectious by means of the "calcium" method of transfection. Cultured human placental cells exposed to EBV DNA of two transforming strains, FF41 and B95, produce virus that is capable of converting normal B lymphocytes into established cell lines. After treatment with EBV (FF41) DNA and EBV (HR-1) DNA the placental cells display antigens associated with the productive viral cycle. The placental cells have not developed foci or other signs of morphologic transformation.

Antigens, Viral↗
PubMed1981

Genetic control of the immune response to staphylococcal nuclease. XI. Effects of in vivo administration of anti-idiotypic antibodies.

The effects of prior treatment with heterologous anti-idiotypic antibodies on the response to staphylococcal nuclease (Nase) have been examined. Previous studies have shown that 100% of A/J mice treated with Nase in completes Freund's adjuvant produce anti-Nase antibodies possessing a characteristic idiotype (Id). Mice treated with anti-Id antibodies followed by Nase produced levels of Id equal to or greater than those of control animals treated with Nase alone. The appearance of Id in treated mice preceded the appearance of anti-Nase activity, and animals treated with anti-Id alone produced high levels of Id without detectable anti-Nase activity. Id expression in such animals could be detected using anti-Id reagents produced in several different species suggesting that it represented true idiotope expression rather than unrelated molecules reactive only with the anti-Id reagent used for initial treatment. Isolation of the nonantigen-binding Id-bearing molecules (Id') showed them to be immunoglobulins bearing the same idiotopes as do anti-Nase antibodies. However, quantitative comparisons of Id levels vs. amount of Id or Id'-bearing immunoglobulin suggested that the nonantigen-binding immunoglobulins bore fewer idiotopes per molecule than did anti-Nase antibodies. Evidence was also obtained for the production of some nonantigen-binding Id-bearing molecules during the normal immune response Nase. These findings are therefore consistent with the existence of a network of Id-anti-Id interactions in the immune response to Nase.

Animals↗
PubMed1981

Genome of a mononucleosis Epstein-Barr virus contains DNA fragments previously regarded to be unique to Burkitt's lymphoma isolates.

We wished to learn whether the genomes of strains of EMB isolated from patients with infectious mononucleosis are consistently distinguishable from those of strains from Burkitt's lymphoma. The genome of a new transforming strains (FF41) of EBV isolated from saliva of a patient with uncomplicated infectious mononucleosis was compared with the DNA of B95-8, the only other available virus from mononucleosis. It had been found previously that B95-8 has a deletion of about 8 Md in the region of the physical map represented by the Eco RI C, Hind III D, and Bam HI I fragments. The W91 and HR-1 isolates for Burkitt's lymphoma are not deleted in this region and it had been proposed that additional information was characteristic of EBV isolates from Burkitt's lymphoma. By means of restriction enzyme analysis, blot hybridization experiments and molecular cloning of FF41 DNA we demonstrate that the deletion found in B95-8 is not present in the new mononucleosis isolate. The FF41 genome contains an extra 8 Md of DNA, represented by Bam HI fragments B', W' and I', which are located in a larger Eco RI C fragment. Thus the genome of this salivary isolate contains DNA that had previously been regarded to be unique to strains from Burkitt's lymphoma. It is therefore unlikely that major insertions or deletions in the EBV genome account for differences in disease manifestation following EBV infection.

Burkitt Lymphoma↗
PubMed1981

Two small RNAs encoded by Epstein-Barr virus and complexed with protein are precipitated by antibodies from patients with systemic lupus erythematosus.

Primate cells harboring the Epstein-Barr virus (EBV) genome synthesize large amounts of two small RNAs:EBER 1 and EBER 2 (EBV-encoded RNA). These RNAs are approximately 180 nucleotides long, possess 5' pppA termini, and lack poly(A). They have different T1 and pancreatic RNase digestion fingerprints. They are not found in normal B lymphocytes, in transformed B lymphocytes that lack EBV DNA, in T lymphocytes transformed by Herpesvirus ateles, or in a variety of other nonlymphoid mammalian cells. Hybridization analyses indicate that EBER 1 and EBER 2 are encoded by the EcoRI-J fragment of EBV (B95-8) DNA. In vivo both RNAs are associated with protein(s), allowing their specific precipitation by the systemic lupus erythematosus-associated antibody anti-La. The La antigen in uninfected mammalian cells consists of a heterogeneous class of small ribonucleoprotein particles, some of whose RNA components exhibit sequence homology with a highly repetitive, interspersed class of human DNA designated the Alu family. Possible functions for EBER 1 and EBER 2 in infection and cell transformation by EBV and their potential relationship to the pathogenesis of systemic lupus erythematosus are discussed.

Antibodies↗
PubMed1981

Resistance of neonatal human lymphoid cells to infection by herpes simplex virus overcome by aging cells in culture.

Mononuclear leukocytes freshly harvested from human umbilical cord blood were refractory to infection by herpes simplex virus (HSV) type 1, whereas cells aged in vitro for a week produced 10- to 100-fold or greater more virus. Increased sensitivity of aged cells to infection by HSV was not dependent on active DNA synthesis by the cells at the time of exposure to HSV. The aging phenomenon was not due to the outgrowth of a specific cell subpopulation that is uniquely susceptible to HSV because increased sensitivity to HSV type 1 occurred in a variety of cell subpopulations enriched or depleted of T lymphocytes, phagocytic cells, or adherent cells. About 25% of aged cells contained virions detectable by electron microscopy, considerably more cells than formed infectious centers. These virions were seen in large activated lymphocytes which are the principal cell subpopulation present in the aged cultures, but virions were also seen in small lymphocytes.

Animals↗
PubMed1981

Dietary vitamin C and uterine cervical dysplasia.

A case-control study of women with cervical abnormalities identified through Pap smears, was conducted in the Bronx, New York, to explore the relationship between nutritional intake and cervical dysplasia. Nutrient intake was estimated from computer analysis of three-day food records and 24-hour recall for 169 study participants (87 cases, 82 controls), including a subset of 49 pairs matched for age, race and parity. Mean vitamin C intake per day from three-day food record for controls was 107 mg, compared to 80 mg for cases (p less than 0.01). Analysis of matched pairs showed similar results; 29% of cases compared to 3% of controls in matched subset had vitamin C intake less than 50% of the recommended daily allowance, yielding a ten-fold increase in risk of cervical dysplasia as estimated by odds ratio (p less than 0.05). Younger age, greater frequency of sexual intercourse and younger age at first intercourse were associated with higher risk of cervical dysplasia. Multiple logistic analyses indicated that low vitamin C intake is an independent contributor to risk of severe cervical dysplasia when age and sexual activity variables are controlled. Approximately 35% of US women in their reproductive years have daily vitamin C intake below 30 mg, and 68% have vitamin C intake below 88 mg. If other studies confirm these findings, it may be important to explore a possible protective role of supplementary vitamin C for women at high risk of cervical cancer.

Adolescent↗
PubMed1981

Mechanisms for Hageman factor activation and role of HMW kininogen as a coagulation cofactor.

Our present concept of the initiating reactions of the intrinsic coagulation pathway is outlined in Figure 5. Although we remain unsure of the etiology shown in Figures 3 and 4, the major function of HMW kininogen is to bind prekallikrein and factor XI in plasma and attach them to surfaces in a conformation that allows activation by HFa. The HMW kininogen--dependent augmentation of the binding of prekallikrein and factor XI to the surface that is seen in plasma (but not buffer systems) would appear to be of lesser importance. Once activated, however, dissociation of kallikrein from the surface allows it to attack adjacent Hageman factor molecules on the same or other particles; this reaction appears to be more rapid than the rate of Hageman factor autoactivation. Thus, the rapid burst of HFa formation seen in normal plasma is kallikrein dependent. It is also dependent upon HMW kininogen, but this appears to be an indirect relationship. The HMW kininogen augments the amount of prekallikrein bound, allows activation to kallikrein, and is needed for kallikrein dissociation from the surface. These three effects all yield a marked increase in the effective ratio of kallikrein/Hageman factor at the surface-fluid interface, and this may be the condition required for rapid HFa formation.

Binding Sites↗