AIDS-related vacuolar myelopathy is not associated with coinfection by human T-lymphotropic virus type I.

Assessment of antibodies against human T-lymphotropic virus type I (HTLV-I) by enzyme-linked immunoassay, immunofluorescence, and Western blot was undertaken in patients with pathologically or clinically diagnosed acquired immunodeficiency syndrome-related vacuolar myelopathy to determine whether this retrovirus could be etiologically implicated in this disorder. No serological evidence for HTLV-I was found in the patients with vacuolar myelopathy, though 1 patient with an atypical myelopathy did have antibodies against HTLV-I.

Nucleosides. 150. Synthesis and some biological properties of 5-monofluoromethyl, 5-difluoromethyl, and 5-trifluoromethyl derivatives of 2'-deoxyuridine and 2'-deoxy-2'-fluoro-beta-D-arabinofuranosyluracil.

A new synthesis of 5-(monofluoromethyl)- and 5-(difluoromethyl)-2'-deoxy-2'-fluoro-beta-D-arabinofuranosyluracil (F-FMAU and F2-FMAU) is reported. 3',5'-Di-O-(tert-butyldiphenyl)silylated thymidine or FMAU was photochemically brominated with NBS to the corresponding alpha-monobromide, which was hydrolyzed to the 5-hydroxymethyl derivative. Further oxidation of the latter with MnO2 afforded the 5-formyluracil nucleoside.

Nucleosides. 146. 1-Methyl-5-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)uracil, the C-nucleoside isostere of the potent antiviral agent 1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)thymine (FMAU). Studies directed toward the synthesis of 2'-deoxy-2'-substituted arabino nucleosides. 6.

The synthesis of 5-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)-1-methyluracil (1, C-FMAU), an isostere of the potent antiviral and antitumor nucleoside 1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)thymine (2'-fluoro-5-methyl-ara-U or FMAU), was achieved. Pseudouridine (2) was converted into 4,5'-anhydro-3'-O-acetyl-2'-O-triflylpseudouridine (4), which was treated with tris(dimethylamino)sulfur (1+) difluorotrimethylsilicate (TASF) to give 4,5'-anhydro-5-(3-O-acetyl-2-deoxy-2-fluoro-beta-D-arabinofuranosyl)-1- methyluracil (5b) in 40% yield. Acid hydrolysis of the 4,5'-anhydro linkage of 5b with Dowex 50 (H+) afforded C-FMAU.

5-substituted 2'-fluoro-arabinonucleosides as potential antiviral agents.

In order to study the electronic effects of 5-substituents of 2'-fluoro-ara-U on antiviral activity, eight nucleosides were synthesized and screened for their activity. Preliminary in vitro studies revealed that 5-thiocyano-, 5-hydroxy- and 5-formyl-ara-U are moderately active against HSV-1, HSV-2 and VZV, but they are also cytotoxic. None of these showed significant activity against CMV.

Comparative methods for detection of thymidine kinase-deficient herpes simplex virus type 1 strains.

Four methods for analyzing viral susceptibility to antiviral substances were compared. In two methods viral products were measured: late viral proteins were measured by an enzyme-linked immunosorbent assay and viral DNA was measured by DNA hybridization. Infectious virus was quantified in the other two assays as the number of plaques and the yield of virus.

Activity of the cytomegalovirus genome in the presence of PPi analogs.

PPi analogs and esters of these were studied for their effect on cytomegalovirus (CMV) multiplication. Five aromatic monoesters of phosphonoformate esterified either in the phosphono or the carboxylic group and two diesters were demonstrated to inhibit CMV DNA synthesis and late viral protein synthesis. In a direct assay, the monoesters but not the diesters inhibited CMV DNA polymerase activity.

Secretion of gamma-interferon at the cellular level. Induction by Epstein-Barr virus.

Using a haemolytic plaque assay for gamma-interferon (IFN-gamma) secretion we found that in vitro Epstein-Barr virus (EBV) exposure of peripheral blood mononuclear cells from EBV immune individuals led to IFN-gamma secretion, which was apparent within 6 h after virus contact and peaked 12-24 h after induction. Live, ultraviolet-light-irradiated and heat-inactivated virions all caused IFN-gamma secretion. In contrast, blood mononuclear cells from EBV non-immune adults or neonates could not be activated to IFN-gamma production by EBV.

Nucleic acid hybridization, a method to determine effects of antiviral compounds on herpes simplex virus type 1 DNA synthesis.

An application of the nucleic acid hybridization technique to screen effects of antiherpes compounds on herpes simplex virus type 1 (HSV-1) DNA synthesis is described. Whole cells are applied to nitrocellulose filters, their DNA is denatured and fixed to the filter. The resulting DNA spots are hybridized to cloned nick-translated HSV-1 DNA and the amount of hybridization is monitored by autoradiography or scintillation counting.

Early interactions between human cytomegalovirus and cells.

Adsorption of cytomegalovirus (CMV) to human fibroblasts was not inhibited by preincubation with other herpes viruses (HSV-1, HSV-2, VZV). Transport of virus to the nucleus was studied using virus labelled with 3H-thymidine. Radioactivity was found in the nucleus 20 minutes after virus had been added to the cells.

Nucleic acid hybridization for measurement of effects of antiviral compounds on human cytomegalovirus DNA replication.

A nucleic acid hybridization technique has been developed to study the effect of different antiviral compounds on the replication of human cytomegalovirus in vitro. One laboratory strain of human cytomegalovirus, Ad. 169, and six clinical isolates were studied.

Adsorption of purified human cytomegalovirus and induction of early antigens in different cells.

Different human and nonhuman cells were assayed for their capacity to absorb human cytomegalovirus (CMV Ad.169) and to support CMV infection in vitro. The CMV adsorptive capacity was assayed by measuring cell-bound radioactivity after addition of purified 3H- or 125I-labeled CMV or by a bioassay for residual infectious virus in the supernatant fluid.

Studies of cytomegalovirus infection in renal allograft recipients. II. Serological response to various viral antigens.

In a prospective study the antibody response to various cytomegalovirus (CMV) antigens was examined in 28 renal allograft recipients. Both primary and secondary infections were investigated. Antibodies against immediate early (IEA) and early antigens (EA) were studied by anti-complement immunofluorescence; IgM and IgG antibodies to nuclear late antigens were differentiated by enzyme-linked immunosorbent assay (ELISA).

Studies of cytomegalovirus infection in renal allograft recipients.

A prospective study of cytomegalovirus (CMV) infections has been carried out in 28 renal graft recipients. The protocol called for frequent blood and urine sampling during the first year after transplantation, but death or graft loss caused earlier termination in nearly half the patients. In this material 5/7 (71%) susceptible patients developed primary infections and 20/21 experienced a secondary infection (95%).

Distribution of tumor-associated antigen CEA and cross-reacting NCA in fetal organs.

The organ distribution of the tumor-associated carcinoembryonic antigen (CEA), and that of the normal tissue component NCA (non-specific cross-reacting antigen) have been investigated in the fetus. Organ extracts from five fetuses between 14 and 21 weeks of age were analysed by radioimmunoassay using specific antisera. CEA was detected in large amounts (800--1,650 ng/g) in fetal colon and in barely detectable amounts in lung and placental tissue.

Increased serum alpha-fetoprotein levels in cytomegalovirus infections.

Among 54 patients with a clinically and serologically verified cytomegalovirus infection, 8 (15%) had raised alpha-fetoprotein (AFP) levels in sera taken after the onset of infection. No raised serum AFP was seen in acute or convalescent sera of 45 patients with other viral diseases, including infectious mononucleosis and acute herpes simplex virus infection. No definite correlation was found between raised AFP and raised S-ALAT values, indicating hepatic damage.

Carcinoembryonic antigen in amniotic fluid.

Carcinoembryonic antigen (CEA), a substance which is known to occur in high amounts in the fetal gut and also in certain tumors of the gastrointestinal tract, has been demonstrated in amniotic fluids from different stages of pregnancy. Radioimmunoassays of CEA in amniotic fluids of 91 normal pregnancies showed a decrease from a mean of 53 ng/ml at 19 weeks to 25 ng/ml at the end of gestation. The CEA activity in amniotic fluid was eluted in the same volume as a standard 125I-CEA on a Sephadex G200 column.