Induction of feline immunodeficiency virus from a chronically infected feline T-lymphocyte cell line

The infection of the feline T-lymphocyte cell line FeT-J with the feline immunodeficiency virus (FIV) Petaluma strain led to the establishment of nonvirus-producing cells. One clone (C15) obtained by limiting dilution was found to express FIV in response to chemical inducers of retroviruses. The chemical treatment of C15 cells led to not only FIV protein synthesis but also an augmentation of viral production. Examination of the C15 cell derivatives obtained by recloning revealed that 10-40% of treated cells constitutively expressed FIV antigens, whereas 100% with expressed FIV antigen in response to the inducer. Chemical induction resulted in more than a 100-fold increase in infectious viral production. The results suggest that a majority of FeT-J cells that are infected with FIV exist in a non-productive state. Establishing a cell line that can be non-productively infected by FIV may help determine the mechanisms of FIV latency.     

Adaptation of feline immunodeficiency virus subtype B strain TM2 to a feline astrocyte cell line (G355-5 cells)

Based on receptor usage during infection, feline immunodeficiency virus (FIV) isolates can be divided into two groups; those that require feline CD134 (fCD134) as a primary receptor in addition to CXCR4 to enter the cells, and those that require CXCR4 only. Most primary isolates, including strain TM2, belong to the former group and cannot infect a feline astrocyte cell line (G355-5 cells) due to a lack of fCD134 expression. In a previous study, we found that G355-5 cells transduced with fCD134 (termed G355-5/fOX40 cells) were susceptible to strain TM2 and the inoculated cells became persistently infected. In this study, we examined the phenotype of the virus prepared from the persistently infected cells (termed strain TM2PI). Intriguingly, strain TM2PI replicated well in na?ve G355-5 cells and the inoculated G355-5 cells (termed G355-5/TM2PI cells) became persistently infected. The infection of TM2PI in G355-5 cells was inhibited by CXCR4 antagonist AMD3100 and TM2PI infected other fCD134-negative, CXCR4-positive cell lines, FeTJ and 3201 cells. Four amino acid substitutions were found in the Env protein of the strain TM2PI when compared with that of the parental strain TM2. Among the substitutions, the Env amino acid position at 407 of TM2PI was substituted to lysine which has been known to be responsible for the FIV tropism for Crandell feline kidney cells. The strain TM2PI will be useful for studying the receptor switching mechanism and FIV pathogenesis in cats.     

High genetic stability of TM1 and TM2 strains of subtype B feline immunodeficiency virus in long-term infection

To know the genetic changes of feline immunodeficiency virus (FIV) in long-term infection in cats, we inoculated three specific pathogen-free cats with FIV isolates and determined a partial env sequence covering the V3-V5 region. In 2 cats infected with subtype B strains TM1 and TM2, only one amino acid change in region V3 was observed at 9 years post infection (y.p.i.), and no nucleotide substitutions were observed between 9 and 10 y.p.i., indicating that these strains are genetically stable. On the other hand, in a cat infected with subtype A strain Petaluma at 8.7 y.p.i., 3 nucleotide insertions (one amino acid insertion) in region V5, and 1 synonymous nucleotide substitution and 2 non-synonymous nucleotide substitutions in region V5, were observed.     

Epidemiology of feline leukaemia and feline immunodeficiency virus infections in the Czech Republic

Commercial serological sets were used for the examination of 727 cats kept in larger towns of the Czech Republic. FeLV antigen and antibodies to FIV were demonstrated in 96 (13.2%) and 42 (5.8%) of the animals, respectively. Seven (0.96%) animals were positive for both FeLV and FIV. Most of the FeLV and/or FIV positive patients were intact rambling males aged 1-4 years. Chronic gastrointestinal and respiratory diseases were found in 54.2% and 43.8% of the FeLV-positive patients, respectively. Chronic urinary tract diseases and generalized lymphadenopathy were found in 47.6% and 45.2% of the FIV-positive patients, respectively. The results of this first survey in the Czech Republic have shown prevalence values and clinical patterns similar to those reported formerly from other European countries.     

Prevalence of feline leukaemia virus and antibodies to feline immunodeficiency virus in cats in the United Kingdom

A representative sample of the pet cat population of the United Kingdom was surveyed. Blood samples from 1204 sick and 1007 healthy cats of known breed, age and sex were tested for antibodies to feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV). The prevalence of FIV was 19 per cent in sick cats and 6 per cent in healthy cats, and the prevalence of FeLV was 18 per cent in sick cats and 5 per cent in healthy cats; both infections were more common in domestic cats than in pedigree cats. Feline immunodeficiency virus was more prevalent in older cats but FeLV was more prevalent in younger cats. There was no difference between the prevalence of FeLV in male and female cats but male cats were more likely to be infected with FIV than female cats. No interaction was demonstrated between FIV and FeLV infections. Of the cats which were in contact with FIV in households with more than one cat, 21 per cent had seroconverted. The prevalence of FeLV viraemia in cats in contact with FeLV was 14 per cent. The clinical signs associated with FIV were pyrexia, gingivitis/stomatitis and respiratory signs, and with FeLV, pyrexia and anaemia. It was concluded that both viruses were significant causes of disease, and that the cats most likely to be infected with FIV were older, free-roaming male cats and for FeLV, younger, free-roaming cats.     

Pilot study of the effect of acemannan in cats infected with feline immunodeficiency virus.

Acemannan, a complex carbohydrate shown to stimulate interleukin-1, tumor necrosis factor alpha and prostaglandin E2 production by macrophages, has also demonstrated antiviral activity in vitro against human immunodeficiency virus, Newcastle disease virus and influenza virus. A pilot study was undertaken to determine acemannan's effect in 49 feline immunodeficiency virus (FIV) infected cats with clinical signs of disease (Stage 3, 4 or 5), 23 of which had severe lymphopenia. Cats received acemannan either by intravenous (Group 1) or subcutaneous (Group 2) injection once weekly for 12 weeks, or by daily oral (Group 3) administration for 12 weeks. Upon entry into the study, cats were randomly assigned to one of the three groups. Laboratory analyses were performed at the beginning of the study and at Weeks 6 and 12. Cats were allowed to continue with a predetermined maintenance regimen of acemannan after completing the 12-week study. Thirteen cats died during the course of treatment. Upon necropsy, the most frequent histopathologic findings were neoplastic, kidney and pancreatic disease. Friedman's two-way ANOVA test showed no significant differences in efficacy among groups administered acemannan by the different routes. Therefore, groups were combined and a signed-ranks test was used to determine changes over time. A significant increase was seen in lymphocyte counts (P < 0.001). Neutrophil counts decreased significantly (P = 0.007), as did incidence of sepsis (P = 0.008). When cats entering with lymphopenia were analyzed separately, a much greater increase in lymphocyte counts was noted (235%) compared with non-lymphopenic cats (42%). A survival rate of 75% was found for all three groups. Thirty-six of 49 animals are alive 5-19 months post-entry. These results suggest that acemannan therapy may be of significant benefit in FIV-infected cats exhibiting clinical signs of disease.     

Effects of cidofovir on cell death and replication of feline herpesvirus-1 in cultured feline corneal epithelial cells

OBJECTIVE: To assess the effect of cidofovir on viability of feline corneal epithelial (FCE) cells, replication of feline herpesvirus (FHV)-1, and virus-induced cytopathic changes. SAMPLE POPULATION: Healthy eyes from 14 recently euthanatized cats. PROCEDURE: Cidofovir at concentrations ranging from 0.05 to 0.000005 mg/mL was added to primary cultures of FCE cells, and cytopathic changes and effects on cell proliferation and cell viability were determined during the subsequent 48 hours. Efficacy of cidofovir (0.02 and 0.05 mg/mL) to prevent in vitro infection of FCE cells with FHV-1 was determined during 72 hours of culture by assessing viral cytopathic effects and viral titers. RESULTS: Cidofovir at concentrations of 0.05, 0.005, and 0.0005 mg/mL significantly reduced mean viable cell counts, and cidofovir at a concentration of 0.05 mg/mL significantly reduced the percentage viability of cultured FCE cells. Minimal cytopathic changes were observed at concentrations of 0.02 and 0.05 mg of cidofovir/mL. Cidofovir at concentrations of 0.05 and 0.02 mg/mL abrogated the cytopathic effects attributable to FHV-1 infection and reduced viral titers from > or =10(14) TCID(50)/mL to < or =10(3.5) TCID50/mL. CONCLUSIONS AND CLINICAL RELEVANCE: Cidofovir in vitro was highly efficacious against FHV-1 infection of a primary culture of FCE cells but had cytostatic effects on cultured cells.     

Inhibitory effects of ribavirin alone or combined with human alpha interferon on feline infectious peritonitis virus replication in vitro

The antiviral activities of ribavirin (1-beta-D-ribofuranosyl-1,2,4-triazole-3-carboxamide; virazole), either alone or in combination with recombinant human leukocyte (alpha) interferon (rHuIFN-alpha), were evaluated against feline infectious peritonitis virus (FIPV) in feline kidney-cell cultures. The 50% inhibitory dose (ID50) of ribavirin for uninfected, rapidly dividing cells was approximately 17 micrograms ml-1 whereas the ID50 for FIPV was 2.5 micrograms ml-1. The therapeutic index (TI) of ribavirin (i.e. the ratio of the minimum cell-toxic dose to minimum virus-inhibitory dose) was 6.8. Although a dose-dependent inhibition of viral infectivity occurred at non-toxic doses, maximum antiviral effects (greater than or equal to 4 log10 reduction in FIPV) occurred at cytotoxic doses. When low or moderate doses of ribavirin were combined with either 10 or 100 U of rHuIFN-alpha ml-1, the resulting antiviral effects were significantly greater than the sum of the observed effects from either ribavirin or rHuIFN-alpha alone. Significant synergistic interactions with rHuIFN-alpha occurred at ribavirin doses of 1, 5, 12.5 and 25 micrograms ml-1. Synergistic combinations of rHuIFN-alpha and ribavirin produced up to an 80-fold or a 200-fold relative increase in FIPV antiviral activities compared with that produced by equivalent doses, respectively, of ribavirin or rHuIFN-alpha alone. In cell growth studies, the addition of either 10 or 100 U of rHuIFN-alpha ml-1 to test doses of ribavirin did not increase the anticellular effect observed with ribavirin alone; seemingly, the potentiation of ribavirin antiviral activity by rHuIFN-alpha was independent of any additive cytotoxic effects. Potentially, synergistic combinations of the two antiviral agents in vivo may decrease the therapeutic dose of ribavirin required for inhibition of FIPV and thus reduce drug toxicity.     

Long-term impact on a closed household of pet cats of natural infection with feline coronavirus, feline leukaemia virus and feline immunodeficiency virus

A closed household of 26 cats in which feline coronavirus (FCoV), feline leukaemia virus (FeLV) and feline immunodeficiency virus (FIV) were endemic was observed for 10 years. Each cat was seropositive for FCoV on at least one occasion and the infection was maintained by reinfection. After 10 years, three of six surviving cats were still seropositive. Only one cat, which was also infected with FIV, developed feline infectious peritonitis (FIP). Rising anti-FCoV antibody titres did not indicate that the cat would develop FIP. The FeLV infection was self-limiting because all seven of the initially viraemic cats died within five years and the remainder were immune. However, FeLV had the greatest impact on mortality. Nine cats were initially FIV-positive and six more cats became infected during the course of the study, without evidence of having been bitten. The FIV infection did not adversely affect the cats' life expectancy.     

Studies of enteric coronaviruses in a feline cell line

Development is reported of a feline cell line which can support the growth of coronaviruses from canine (CCV), feline (FIPV) and porcine (TGEV) species. The cell culture has been serially transferred over 100 times and has retained its initial growth requirements, proliferative capacity and morphologic features. Each virus had specific growth characteristics in this cell culture although all produced a similar CPE and plaques under agar. Cross neutralization studies demonstrated a two-way relationship between TGEV and CCV and between TGEV and FIPV, whereas a one-way relationship was demonstrated between CCV and FIPV.     

Prevalence of feline immunodeficiency virus in submissions of feline serum to a diagnostic laboratory in Atlantic Canada

The purpose of this project was to identify the prevalence of feline immunodeficiency virus (FIV) in the Atlantic region of Canada, and to determine possible associations between FIV serological status and breed, sex, and age. Feline serum samples (671) submitted to the Prince Edward Island Diagnostic Services — Atlantic Veterinary College laboratory between January 1, 1988 and July 30, 1989 were considered eligible for this study. The majority of samples originated from Prince Edward Island (607). Testing was performed in duplicate using commercial 96-well enzyme-linked immunosorbent assay test kits for FIV antibody. Results included a seropositive rate of 7.6% for all submissions. Mean age of FIV-seropositive cats was eight years. There was an increasing risk of FIV-seropositive status associated with age. Prevalence of FIV among intact males was significantly higher (odds ratio = 2.59) than other gender categories. The principal conclusion of this study was that FIV is present in cats of the Atlantic provinces, and that its associations and prevalence are consistent with those found in other North American epidemiological studies.     

Phylogenetic characterisation of naturally occurring feline immunodeficiency virus in the United Kingdom

Feline immunodeficiency virus (FIV) is a significant pathogen of domestic and non-domestic felids worldwide. In domestic cats, FIV is classified into five distinct subtypes (A-E) with subtypes A and B distributed most widely. However, little is known about the degree of intrasubtype viral diversity and this may prove critical in determining whether monovalent vaccines are likely to protect against FIV strains within a single subtype. Here, we characterise novel env sequences from 47 FIV strains recovered from infected cats in the United Kingdom and its environs. Phylogenetic analyses revealed that all bar one sequence belonged to subtype A, the predominant subtype in Western Europe. A single sequence was identified as a likely subtype A/C recombinant, intriguing given that subtype C does not appear to exist in either the UK or North Western Europe and suggestive of a recombination event predating its introduction into the UK. Subtype A strains from the UK were not significantly differentiated from representative subtype A isolates found elsewhere suggesting multiple introductions of FIV into the country. Divergence among isolates was comparable to that observed for subtype A isolates worldwide, indicating that FIV in the UK covers the full spectrum of subtype A diversity seen globally. This study demonstrates that while subtype A is predominant in the UK, novel introductions may result in the emergence of novel subtypes or intersubtype recombinants, potentially circumventing vaccine strategies. However, the dominance of subtype A suggests that the development of a regional or subtype-specific protective vaccine for the UK could be achievable.