Feline immunodeficiency virus: a neurotropic lentivirus.

We have investigated the neurotropism of feline immunodeficiency virus (FIV) in naturally and experimentally infected cats. Antibodies to FIV were detected in the cerebrospinal fluid (CSF) of 9 of 10 naturally infected cats, and the virus was cultured from the CSF of 5 of 9 of these cats. After experimental intrathecal or intra-bone-marrow inoculation, FIV antibodies were detected in CSF, as were CSF pleocytosis, increased IgG concentration, and elevated CSF IgG index. Brain lesions consisting of perivascular mononuclear cell infiltrates and both glial nodules and diffuse gliosis were observed in the midbrain and thalamus 7 months after inoculation. Virus was recovered by primary culture of brain tissue from several brain regions (cerebral cortex, caudate nucleus, midbrain, cerebellum, rostral and caudal brainstem) but was not recovered from choroid plexus. In vitro, FIV infected primary cultures of feline astrocytes and brain macrophages. Infection of astrocytes resulted in early syncytium formation, production of infectious virions, and eventual cell death. In brain macrophages, FIV infection was noncytopathic. Productive infection of feline neurons or oligodendrocytes was not observed. We conclude that FIV is a neurotropic lentivirus and that FIV infection of feline CNS may serve as a useful model for study of human immune deficiency virus infection of the human CNS.

FeLV-FAIDS-induced immunodeficiency syndrome in cats.

Findings are reviewed, relevant to elucidation of the pathogenic, genetic and biochemical properties of a single, genetically heterogeneous isolate of feline leukemia virus (FeLV-FAIDS) shown to induce fatal immunodeficiency disease in nearly 100% of inoculated cats. Hypotheses are suggested which pertain to the mechanism of T-cell killing by this virus, and which extrapolate findings in the FeLV-FAIDS animal model to AIDS induced in humans by human immunodeficiency virus (HIV).

Central nervous system infection associated with anaerobic bacteria in two dogs and two cats.

Central nervous system (CNS) infection caused by anaerobic bacteria (including Bacteroides, Fusobacterium, Peptostreptococcus, and Eubacterium) was diagnosed in two dogs and two cats. In one dog there was extensive meningomyeloencephalitis, presumably the result of hematogenous spread of bacteria from lung abscesses and bacterial endocarditis. Subdural empyema of unknown origin was found in a second dog and two cats. Clinical signs in all four animals included mental depression and focal neurologic deficits, without fever.

Central nervous system toxicosis associated with metronidazole treatment of dogs: five cases (1984-1987).

Metronidazole, administered to 5 dogs for periods ranging from 3 to 14 days, was associated with acute development of CNS dysfunction. Metronidazole dosage ranged from 67.3 to 129.0 mg/kg of body weight/d. Clinical signs of toxicosis began with anorexia and intermittent vomiting and progressed rapidly to include pronounced, generalized ataxia and vertical, positional nystagmus. These signs were consistent with lesions of the vestibular nuclei and/or cerebellum. High CSF protein content was detected in 2 of 3 dogs from which CSF was collected. Two dogs were euthanatized because of severe neurologic dysfunction. Three dogs improved slowly and recovered completely over several months. These findings suggest that currently recommended dosages of metronidazole may be too high for some dogs.

Posttranslational modifications distinguish the envelope glycoprotein of the immunodeficiency disease-inducing feline leukemia virus retrovirus.

The envelope glycoprotein (gp70) of a molecularly cloned, replication-defective feline leukemia virus (FeLV-FAIDS clone 61C) carries determinants for induction of fatal immunodeficiency disease, whereas the gp70 of its companion replication-competent, probably parent virus (clone 61E) does not. Immunoprecipitation analysis of the extracellular glycoproteins of 61E and EECC, a replication-competent viral construct composed of the 61C env and 3' long terminal repeat fused to the 61E gag-pol genes, demonstrated that the gp70 of EECC could be distinguished from that of 61E by both feline immune serum and a murine monoclonal antibody. Molecular weights of both the envelope precursor polyprotein (gp80) and the mature extracellular glycoprotein (gp70) of 61E were smaller than the corresponding proteins from the pathogenic EECC. Both the molecular weight disparity and monoclonal antibody discrimination of the two gp80s were abolished by inhibition of envelope protein glycosylation with tunicamycin, whereas the apparent gp70 size differences were resolved by enzymatic removal of N-linked oligosaccharides. Pulse-chase studies in EECC-infected cells demonstrated that processing of gp80 to gp70 was delayed and that this retardation of envelope glycoprotein processing could be simulated in 61E-infected cells by treatment with the glucosidase inhibitor N-methyldeoxynojirimycin, a compound that causes retention of oligosaccharides in the high-mannose form. The resultant 61E gp70 then could be recognized by sera from EECC-immunized cats. The presence of a higher content of sialic acid on the apathogenic 61E gp70 indicated that oligosaccharides of 61E and EECC gp70 were processed differently. These data suggested that the unique biochemical properties which distinguish the envelope glycoproteins of the FeLV-FAIDS variant from its companion apathogenic parent virus were responsible for T-cell cytopathicity and induction of immunodeficiency disease. Further biochemical characterization of these glycoproteins should be useful in understanding the pathogenic mechanisms of immunodeficiency disease induced by retroviruses.

Cell-specific envelope glycosylation distinguishes FIV glycoproteins produced in cytopathically and noncytopathically infected cells.

Feline immunodeficiency virus (FIV) infection induces syncytium formation and cell death in primary feline astrocyte cultures but persistently and noncytopathically infects Crandell feline kidney cells (CrFK). Because viral envelope glycoproteins are implicated in cell fusion events we evaluated the astrocyte-produced FIV surface glycoprotein for properties that might distinguish it from that produced in CrFK cells. The surface glycoprotein from astrocytes migrated faster on SDS-PAGE and contained more Endo H-sensitive oligosaccharides than that from CrFK, although the precursor and deglycosylated envelope glycoproteins from both cells were the same size. Castanospermine treatment of infected astrocytes, which blocks glucose trimming from oligosaccharide side chains of glycoproteins, both obliterated the mobility difference between astrocyte- and CrFK-produced FIV surface glycoproteins and prevented syncytium in infected astrocyte cultures. These results demonstrate the importance of the infected cell type in viral envelope protein glycosylation and implicate cell type-specific carbohydrate structures on retroviral glycoproteins as mediators of cell fusion.

Characterization and significance of delayed processing of the feline leukemia virus FeLV-FAIDS envelope glycoprotein.

FeLV-FAIDS, an immunodeficiency-inducing isolate of feline leukemia virus, is composed of a pathogenic but replication-defective genome (molecular clone 61C) and a replication-competent but non-immunodeficiency-inducing variant genome (molecular clone 61E). The chimeric virus EECC, composed of the 5' gag-pol of 61E fused to the env-3' LTR of 61C, also induces immunodeficiency. The 61C (or EECC) gp80 can be distinguished from that of 61E on the basis of antigenic recognition, size, and rate of posttranslational processing. We found that the nascent precursor polypeptides of the two viruses were the same size; however, the 61E gp80 rapidly shifted to a smaller size and was subsequently cleaved to gp70, whereas EECC gp80 maintained its nascent size and was cleaved to gp70 only after a prolonged time. Endo-beta-N-acetyl glucosaminidase H and N-glycanase digestions of newly formed glycoproteins resulted in a similar banding pattern for both viruses, indicating that both contained the same number of oligosaccharide side chains and that all of these were high mannose sugars. The metabolic inhibitors of glycosylation, castanospermine or N-methyldeoxynojirimycin, prevented both the rapid trimming of 61E gp80 and its cleavage to gp70. Treatment with mannosidase inhibitors, however, did not affect 61E gp80 processing or size, suggesting that retention of glucose residues on EECC was responsible for these distinguishing properties of the glycoprotein. The pathological consequence of aberrant viral glycoprotein processing was evaluated in feline 3201 T lymphocytes, which are infectable by both 61E and EECC but are killed only by EECC. As in fibroblasts, the EECC glycoprotein produced in lymphocytes was larger, antigenically distinct, and processed more slowly than was the glycoprotein of 61E. Castanospermine treatment of 61E-infected 3201 T cells, however, not only abrogated the antigenic differences between the 61E and EECC glycoproteins but also resulted in a cytopathic effect. Our results suggest that (i) intracellular accumulation of EECC envelope glycoprotein may occur consequent to retention of glucose residues on carbohydrate side chains and (ii) a strong correlation exists between delayed glycoprotein processing and cytopathicity in FeLV-FAIDS-infected T lymphocytes.

Mutations within a putative cysteine loop of the transmembrane protein of an attenuated immunodeficiency-inducing feline leukemia virus variant inhibit envelope protein processing.

A replication-defective feline leukemia virus molecular clone, 61B, has been shown to cause immunodeficiency in cats and cytopathicity in T cells after a long latency period when coinfected with a minimally pathogenic helper virus (J. Overbaugh, E. A. Hoover, J. I. Mullins, D. P. W. Burns, L. Rudensey, S. L. Quackenbush, V. Stallard, and P. R. Donahue, Virology 188:558-569, 1992). The long-latency phenotype of 61B has been mapped to four mutations in the extracellular domain of the envelope transmembrane protein, and we report here that these mutations cause a defect in envelope protein processing. Immunoprecipitation analyses demonstrated that the 61B gp85 envelope precursor was produced but that further processing to generate the surface protein (SU/gp70) and the transmembrane protein (TM/p15E) did not occur. The 61B precursor was not expressed on the cell surface and appeared to be retained in the endoplasmic reticulum or Golgi apparatus. Two of the four 61B-specific amino acid changes are located within a putative cysteine loop in a region of TM that is conserved among retroviruses. Introduction of these two amino acid changes into a replication-competent highly cytopathic virus resulted in the production of noninfectious virus that exhibited an envelope-protein-processing defect. This analysis suggests that mutations in a conserved region within a putative cysteine loop affect retroviral envelope protein maturation and viral infectivity.