Feline leukaemia virus status of Australian cats with lymphosarcoma

OBJECTIVE: To determine the FeLV status of sera and tumours from Australian cats with lymphosarcoma in relation to patient characteristics, tumour characteristics (tissue involvement, histological grade and immunophenotype), haematological and biochemical values. DESIGN: Prospective study of 107 client-owned cats with naturally-occurring lymphosarcoma. PROCEDURE: An ELISA was used to detect FeLV p27 antigen in serum specimens collected from cats with lymphosarcoma. A PCR was used to detect FeLV DNA in formalin-fixed, paraffin-embedded tissue sections containing neoplastic lymphoid cells. The PCR was designed to amplify a highly conserved region of the untranslated long terminal repeat of FeLV provirus. RESULTS: Only 2 of 107 cats (2%), for which serum samples were available, were FeLV-positive on the basis of detectable p27 antigen in serum. In contrast, 25 of 97 tumours (26%) contained FeLV DNA. Of the 86 cats for which both PCR and ELISA data were available, 19(22%) had FeLV provirus in their tumours but no detectable circulating FeLV antigen in serum, while 2 (2%) had FeLV provirus and circulating FeLV antigen. FeLV PCR-positive/ELISA-negative cats (19) differed from PCR-negative/ELISA-negative cats (65) in having fewer B-cell tumours (P = 0.06), more non B-/non T-cell tumours (P = 0.02) and comprising fewer non-Siamese/Oriental pure-bred cats (P = 0.03). CONCLUSIONS: The prevalence of FeLV antigen or provirus was considerably lower in our cohort of cats compared with studies of lymphosarcoma conducted in the Northern hemisphere. This suggests that factors other than FeLV are important in the development of lymphosarcoma in many Australian cats. No firm conclusions could be drawn concerning whether FeLV provirus contributed to the development of lymphosarcoma in PCR-positive/ELISA-negative cats.     

Therapy for Australian cats with lymphosarcoma

OBJECTIVE: To determine the response of Australian cats with lymphosarcoma to chemotherapy and/or surgery in relation to patient and tumour characteristics, haematological and serum biochemical values and retroviral status. DESIGN: Prospective study of 61 client-owned cats with naturally-occurring lymphosarcoma subjected to multi-agent chemotherapy and/or surgery. PROCEDURE: An accepted chemotherapy protocol utilising l-asparaginase, vincristine, cyclophosphamide, doxorubicin, methotrexate and prednisolone was modified and used to treat 60 cats with lymphosarcoma. Clinical findings were recorded before and during therapy. As far as practical, cases were followed to death, euthanasia or apparent cure. Owner satisfaction with the results of chemotherapy was determined using a questionnaire sent after the completion of chemotherapy. RESULTS: One cat, with lymphosarcoma limited to a single mandibular lymph node, was treated using surgery alone and was cured. The other 60 cats were treated using multi-agent chemotherapy, although seven cats with localised intestinal, ocular and subcutaneous lesions had these lesions partially (2 intestinal lesions) or completely (2 eyes, 2 intestinal lesions and a cluster of regional lymph nodes) resected prior to starting chemotherapy. The median survival time for these 60 cats was 116 days. Of the 60 cats, 48 rapidly went into complete remission following the administration of 1-asparaginase, vincristine and prednisolone (complete remission rate 80%) and these cats had a median survival of 187 days. Three cats were censored from further analysis as their long-term survival data were uninterpretable because they died of causes unrelated to lymphosarcoma or were prematurely lost to follow-up. Twenty cats were classed as 'long-term survivors' based on survival time in excess of one year and at least 14 were 'cured' based on the absence of physical evidence of lymphosarcoma 2-years after initiating treatment. In other words, of the 48 cats that reached complete remission, in excess of 29% were 'cured'. Despite detailed analysis, few meaningful prognostic indicators based on patient or tumour characteristics were identified, although long-term survivors were more likely to be less than 4-years (P= 0.04) and to have tumours of the T-cell phenotype (P= 0.06). Excluding the one FeLV ELISA-positive cat with mediastinal LSA, 7 of 9 cats less than 4 years-of-age were long-term survivors (median survival time >1271 days). There was a strong association between achieving complete remission and long-term survival (P = 0.003). On the basis of 27 replies to a questionnaire, owners were generally very satisfied with the response to chemotherapy, irrespective of the survival time of the individual patient. Eighty five percent of owners expressed complete satisfaction with their decision to pursue chemotherapy and 70% believed their cat's health status improved during the first 2-weeks of treatment. Importantly, 78% of owners considered that chemotherapy required a very substantial time commitment on their part. CONCLUSIONS: It was possible to cure approximately one quarter of cats with lymphosarcoma using sequential multi-agent chemotherapy and/or surgery. FeLV-negative cats younger than 4 years (typically with mediastinal lymphosarcoma) had a particularly favourable prognosis. The decision to embark on chemotherapy should be based on the results of induction chemotherapy with l-asparaginase, vincristine and prednisolone, as the response to this was a good predictor of long-term survival. Cats surviving the first 16 weeks of chemotherapy generally enjoyed robust remissions (in excess of 1 year) or were cured of their malignancy.     

Prevalences of feline leukaemia virus and feline immunodeficiency virus infections in cats in Sydney

Objective To determine prevalences of feline leukaemia virus (FeLV) and feline immunodeficiency virus (FIV) infections in ‘healthy’ cats that, through acute misadventure or other circumstance, were presented to veterinary practitioners. Prevalences of FeLV and FIV in this population were compared to those in a population of predominantly sick cats. Design and procedures Serum specimens were obtained over a 2-year period from 200 cats oldeer than 1 year of age presented to veterinary clinics for routine procedures, including cat fight injuries or abscesses, vehicular trauma, neutering, dental scaling, vaccination, grooming or boarding. An additional 894 sera were obtained over approximately the same period from specimens submitted by veterinarians to a private clinical pathology laboratory, mainly from sick cats suspected of having immune dysfunction, but including some sera from healthy cats being screened prior to FeLV vaccination. FIV antibody and FeLV antigen were detected in samples using commercial enzyme immunoassays. Results Amongst 200 ‘healthy’ cats, the prevalence of FeLV infection was 0 to 2%, and the prevalence of FIV was 6.5 to 7.5%, depending on the stringency of the criteria used to define positivity. FIV infection was significantly more prevalent in cats which resided in an inner city environment (P = 0.013). Of the 894 serum specimens submitted to the laboratory by practitioners, 11/761 (1.4%) were FeLV positive, while 148/711 (20.8%) were FIV positive. The prevalence of FIV was significantly higher in these predominantly ‘sick’ cats than in cats seen for routine veterinary procedures (P < 0.00001), while there was no difference in the prevalence of FeLV (P = 0.75) Conclusions The prevalence of FeLV and FIV in healthy cats may have been substantially overestimated in some previous Australian surveys. FeLV infection would appear to be a rare cause of disease in Australian cats. The higher prevalence of FIV positivity in sick as opposed to healthy cats infers that FIV infection contributes to the development of disease.     

Clinical and anatomical features of lymphosarcoma in 118 cats

Objective To determine patients' characteristics and anatomical distribution of lesions in cats with lymphosarcoma. Design Prospective multi-institutional study of naturally occurring feline lymphosarcoma. Methods Veterinarians in Sydney were provided with free diagnostic laboratory services for suspect cases of feline lym-phosarcoma. Lymphosarcoma was diagnosed based on physical findings, radiographic and/or ultrasonographic images and results of cytological or histopathological examination. When owners were not interested in pursuing an antemortem diagnosis, suspect cases were collected for necropsy. Patients' characteristics and physical findings were recorded. A modified scheme for anatomical classification of lesions was devised including a ‘mixed’ category for cases which involved two or more anatomical forms. Results One hundred and eighteen cases were accrued over an 18 month period. The median age was 120 months and range 5 to 212 months. Age distribution was bimodal, with a small peak for cats less than 24 months, and a normal distribution centred on 97 to 120 months. Eighty cats were domestic crossbreds, 22 were Siamese or Oriental cats (including crosses), 6 were Burmese, 5 were purebred longhairs and the remaining 5 were one of a number of purebred shorthaired breeds. In comparison to 1017 consecutive cases admitted to our hospital for conditions other than lymphosarcoma, Siamese/Oriental cats were over-represented amongst lymphosarcoma cases (P = 0.0006). Male cats were also over-represented, accounting for 72 of 118 cases (P = 0.05). Abdominal lymphosarcoma was the most common anatomical form (43 cats), followed by mixed (39), nodal (20), mediastinal (9) and atypical (involving non-lymphoid organs, 7) forms. When analysed for specific organ involvement, 29 (25%) had mediastinal involvement, 71 (60%) had abdominal involvement including 60 (51%) with involvement of the intestinal tract and/or mesenteric lymph nodes and 36 (31%) with bilateral renal involvement, and 47 (40%) had peripheral lymph node involvement. No case of primary lymphoid leukaemia was identified. A noticeable subgroup of cats younger than 24 months had involvement of the anterior mediastinum with or without concurrent enlargement of cervical or axillary lymph nodes; Siamese/Oriental cats were over-represented in this subgroup. Among cases with nodal involvement, lymph nodes of the head and neck were frequently involved, mandibular nodes most commonly, followed by superficial cervical nodes. In seven cases a solitary node was affected. Conclusions Compared with similar surveys overseas, our cats were older and male cats were over-represented. There was a notable subgroup of young cats with mediastinal involvement. Siamese/Oriental cats were over-represented in this subgroup as well as in the larger population of cats with lymphosarcoma. Compared with overseas surveys, renal involvement, mixed cases and atypical cases (including nasal lymphosarcoma) were more common. A new subcategory of nodal lymphosarcoma, with involvement restricted to node(s) of head and neck, was identified.     

Characterisation of lymphosarcomas in Australian cats using polymerase chain reaction and immunohistochemical examination

Objective To examine tumour tissue of cats with lymphosarcoma for the presence of feline leukaemia virus and feline immunodeficiency virus and analyse the immunophenotype of the tumours.
Design A retrospective study of feline lymphosarcoma cases.
Methods Formalin-fixed, paraffin-embedded tumour tissue of 14 feline lymphosarcomas was examined for the presence of feline leukaemia virus and feline immunodeficiency virus by polymerase chain reaction and immunohistochemistry. Using polyclonal and monoclonal antibodies against T and B lymphocytes, the phenotypic expression of the tumours was characterised.
Results No feline leukaemia virus antigen or proviral sequences were detected. Feline immunodeficiency virus proviral sequences were detected in two cases by polymerase chain reaction. Immunophenotyping of all 14 cases resulted in seven cases being classified as B-cell phenotype, four as T-cell phenotype, and the remaining three undetermined.
Conclusions In contrast to previous reports overseas, our results suggest that feline leukaemia virus infection appears to be an infrequent cause of lymphosarcoma in the cats that were necropsied. Feline immunodeficiency virus may have a role in lymphomagenesis. The potential role of feline immunodeficiency virus needs to be explored in more depth. Compared with most previous reports, B-cell tumours were more common than T-cell tumours in this series of cats.     

Prevalence and risk factors for cats testing positive for feline immunodeficiency virus and feline leukaemia virus infection in cats entering an animal shelter in New Zealand

Abstract

AIMS

To estimate the prevalence of cats testing positive for antibodies to feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV) antigens in domestic cats entering a New Zealand animal shelter, based on a commercial point-of-care ELISA, to identify risk factors associated with cats testing positive, and to compare the results obtained from the ELISA with those obtained using PCR-based testing.     

Diagnosing feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV) infection: an update for clinicians

With the commercial release in Australia in 2004 of a vaccine against feline immunodeficiency virus (FIV; Fel‐O‐Vax FIV®), the landscape for FIV diagnostics shifted substantially. Point‐of‐care (PoC) antibody detection kits, which had been the mainstay for diagnosing FIV infection since the early 1990s, were no longer considered accurate to use in FIV‐vaccinated cats, because of the production of vaccine‐induced antibodies that were considered indistinguishable from those produced in natural FIV infections. Consequently, attention shifted to alternative diagnostic methods such as nucleic acid detection. However, over the past 5 years we have published a series of studies emphasising that FIV PoC test kits vary in their methodology, resulting in differing accuracy in FIV‐vaccinated cats. Importantly, we demonstrated that two commercially available FIV antibody test kits (Witness? and Anigen Rapid?) were able to accurately distinguish between FIV‐vaccinated and FIV‐infected cats, concluding that testing with either kit offers an alternative to PCR testing. This review summarises pertinent findings from our work published in a variety of peer‐reviewed research journals to inform veterinarians (particularly veterinarians in Australia, New Zealand and Japan, where the FIV vaccine is currently commercially available) about how the approach to the diagnosis of FIV infection has shifted. Included in this review is our work investigating the performance of three commercially available FIV PoC test kits in FIV‐vaccinated cats and our recommendations for the diagnosis of FIV infection; the effect of primary FIV vaccination (three FIV vaccines, 4 weeks apart) on PoC test kit performance; our recommendations regarding annual testing of FIV‐vaccinated cats to detect ‘vaccine breakthroughs’; and the potential off‐label use of saliva for the diagnosis of FIV infection using some FIV PoC test kits. We also investigated the accuracy of the same three brands of test kits for feline leukaemia virus (FeLV) diagnosis, using both blood and saliva as diagnostic specimens. Based on these results, we discuss our recommendations for confirmatory testing when veterinarians are presented with a positive FeLV PoC test kit result. Finally, we conclude with our results from the largest and most recent FIV and FeLV seroprevalence study conducted in Australia to date.     

Combination chemotherapy in feline lymphoma: treatment outcome, tolerability, and duration in 23 cats

BACKGROUND: Different chemotherapy regimes have been described for feline lymphoma with varying outcomes. HYPOTHESIS: In cats with lymphoma, a long-term, multiagent chemotherapy protocol will be effective and carry acceptable toxicity. ANIMALS: Twenty-three cats with histologically or cytologically confirmed diagnosis of lymphoma. METHODS: Prospective, single-arm clinical trial in which cats were treated with a chemotherapy protocol consisting of a cyclic combination of l-asparaginase, vincristine, cyclophosphamide, doxorubicin, methotrexate, and prednisolone with a planned total treatment time of 122 weeks. RESULTS: Complete remission (CR) rate was 74% (n = 17). Fourteen percent of cats attained partial remission (PR). Median duration of first CR was 264 days (range, 45-2,485 days). Six-month, 1-, and 2-5-year remission rates were 75, 50, and 34%, respectively. Duration of PR ranged between 23 and 63 days. Median survival in cats with CR was 296 days (range, 50-2,520 days). Six-month, 1-, 2-, and 3-5-year survival rates in cats with CR were 82, 47, 34, and 27%, respectively. Survival of cats achieving PR ranged between 38 and 120 days. Of the analyzed variables, only anatomical location had a significant influence on remission duration (P=.022). Actual median treatment time in cats with CR was 128 days (18 weeks). Hematologic and gastrointestinal toxicosis was infrequent and mostly low grade. CONCLUSIONS AND CLINICAL IMPORTANCE: In this population of cats with lymphoma, chemotherapy was effective. With infrequent and mostly low-grade toxicosis, tolerability of the protocol may be considered good.     

Serological differentiation of FIV-infected cats from dual-subtype feline immunodeficiency virus vaccine (Fel-O-Vax FIV) inoculated cats

Feline immunodeficiency virus (FIV) vaccine, Fel-O-Vax FIV, was released for sale in the US in 2002. The antibodies of vaccinated cats interfere with serological assays by currently available FIV diagnostic kits. In this study, we investigated whether it is possible to distinguish serologically cats vaccinated with Fel-O-Vax FIV from cats experimentally or naturally infected with FIV. A total of 153 sera taken from 97 cats were used as serum samples. Enzyme linked immunosorbent assay (ELISA) was performed using whole FIV antigen and formalin treated whole FIV antigen, recombinant-gag (r-gag) antigen, and transmembrane (TM) peptide. Statistical analysis was performed using ELISA optical density (O.D.) values obtained with each antigen as variables. Except for the ELISA O.D. values obtained with r-gag antigen, a significant difference in ELISA O.D. values was observed between the vaccinated and the infected groups. However, it was not possible to distinguish both groups unequivocally. Using discriminant analysis, it was possible to distinguish the two groups with an accuracy of 97.1% with two discriminating variables (ELISA O.D. values obtained with formalin treated whole FIV antigen, and TM peptide), 97.8% with three discriminating variables (ELISA O.D. values obtained with whole FIV antigen, formalin treated whole FIV antigen, and TM peptide). Therefore, it was considered possible to distinguish cats vaccinated with Fel-O-Vax FIV from FIV-infected cats by ELISA using two types of antigens including formalin treated whole FIV antigen and TM peptide, or three types of antigens including formalin treated whole FIV antigen, TM peptide and whole FIV antigen.     

猫免疫缺陷病毒在医学中的应用

猫免疫缺陷病毒（Feline immunodeficiency virus，FIV）是一种主要感染猫的反转录病毒。FIV与人的免疫缺陷病毒（Human immunodeficiency virus，HIV）有许多相似性，可作为HIV的研究模型，如测量FIV病毒在动物体内的感染曲线，可为了解HIV在人体内的传播细节提供蓝图；利用FIV偏好感染发展中的神经系，可以帮助人们了解艾滋病（Acquired immure deficiency syndrome，AIDS）对神经系统的致病机理。FIV疫苗的研究获得成功，可为HIV疫苗的研制提供提示。FIV在基因治疗方面有重要的应用价值，目前应用FIV传递CFTR（cystic fibrosis transmembrane conductance regulator）的cDNA到呼吸系统表皮细胞治疗囊肿性纤维化（cysticfibrosis，CF）的研究已经取得了很大进展。     

猫免疫缺陷病毒在医学中的应用

猫免疫缺陷病毒(Feline immunodeficiency virus,FIV)是一种主要感染猫的反转录病毒.FIV与人的免疫缺陷病毒(Human immunodeficiency virus,HIV)有许多相似性,可作为HIV的研究模型,如测量FIV病毒在动物体内的感染曲线,可为了解HIV在人体内的传播细节提供蓝图;利用FIV偏好感染发展中的神经系,可以帮助人们了解艾滋病(Acquired immure deficiency syndrome,AIDS)对神经系统的致病机理.FIV疫苗的研究获得成功,可为HIV疫苗的研制提供提示.FIV在基因治疗方面有重要的应用价值,目前应用FIV传递CFTR(cystic fibrosis transmembrane conductance regulator)的cDNA到呼吸系统表皮细胞治疗囊肿性纤维化(cystic fibrosis,CF)的研究已经取得了很大进展.     

Molecular subtyping of feline immunodeficiency virus from cats in Melbourne

OBJECTIVE: To determine the subtypes of feline immunodeficiency virus (FIV) present in the domestic cat population in Melbourne. METHODS: Blood samples were collected from 42 cats that had serum antibodies against FIV. DNA was extracted and subjected to polymerase chain reaction (PCR) to amplify variable regions of the envelope (env) and group specific antigen (gag) genes of FIV. PCR products were directly sequenced or sequenced after cloning when direct sequencing yielded ambiguous results. Phylogenetic analysis was performed and comparisons made with representative sequences of different subtypes. RESULTS: The variable region of the env gene was successfully amplified by PCR from 41 of the 42 cats. All 41 were found to cluster with subtype A env sequences. The variable region of the gag gene was successfully amplified by PCR from all 42 cats. Forty-one were found to cluster with subtype A gag genes and one was found to cluster with subtype B sequences, suggesting that it may be derived from a recombinant env A/gag B virus. CONCLUSIONS: Subtype A is the predominant FIV type in Melbourne, although a subtype A/B recombinant was identified in the population of FIV positive cats. These results of env gene analysis were similar to those in a previous Australian study, suggesting that subtype A predominates in Australia. The results of the gag gene analysis show the importance of analysing multiple areas of the FIV genome when assigning FIV subtypes. Comparison with other major urban centres may provide useful information about the phylogenic diversity of FIV in Australia.     

Evaluation of barcoded magnetic bead–based immunoassays for the simultaneous detection of feline leukemia virus antigen and antibody against feline immunodeficiency virus

Testing platforms that leverage automation, require minimal sample volume, and enable various tests to be performed simultaneously on a single sample have the potential to improve workflow and efficiency in veterinary diagnostic laboratories. We evaluated a barcoded magnetic bead (BMB) technology using established immunoassays for detection of feline leukemia virus (FeLV) p27 antigen and antibody against feline immunodeficiency virus (FIV). Analytical sensitivity, limit of blank, and limit of detection were used to establish a functional sensitivity of 1.00 ng/mL of inactivated FeLV antigen and 35.7 ng/mL of anti-FIV monoclonal antibody. Common interferents, such as hemoglobin, lipid, and bilirubin, were not found to interfere with the performance of the assay. Intra- and inter-assay CVs were <13% for both assays using manufactured samples. Using a set of 116 feline samples, the diagnostic accuracy of our multiplex assay was 100% compared to reference assays. Performance in a convenience set of 1,000 feline samples submitted to a commercial diagnostic laboratory revealed a proportion of positive results of 1.3% for FeLV and 3.7% for FIV. BMB technology should enable rapid screening of samples for various markers in a single immunoassay well.     

Association between feline immunodeficiency virus antibodies and host characteristics in Finnish cats.

Toward the end of 1989 the largest private veterinary laboratory in Finland (Vet/lab) began using a commercial combined ELISA test for Feline Immunodeficiency Virus (FIV) antibodies and Feline Leukemia Virus (FeLV) antigens (Cite Combo). The overall proportion of FIV seropositive feline samples was 5% during the 22 month study period. The number of tests performed increased slowly while the positive test results decreased with time (7% in 1990 and 4% in 1991). The decrease in prevalence was assumed to reflect a change in the sample population rather than an actual change in the general cat population. There were more symptomatic and domestic cats tested in 1990 than 1991. The lower-risk groups in the second year of the study may simply be an indication that the cat owners became more aware of FIV and the motivation to send samples switched from the veterinarian's interest to diagnose the disease in a symptomatic cat to the owner's interest to survey their cats for possible FIV infection. In a multivariable analysis, breed, symptoms, age and sex were associated with the risk of FIV seropositivity. The risk increased faster with age in males than in females (i.e., the age effect was not constant between sexes). The cats with symptoms had a higher risk than those without symptoms and non-purebred cats were at a higher risk than purebred cats. FeLV infection was not associated with FIV.     

Feline immunodeficiency virus infection: a valuable model to study HIV-1 associated encephalitis

Feline immunodeficiency virus (FIV), like human immunodeficiency virus (HIV)-1, is a neurotropic lentivirus and is associated with neuropathology in natural and experimental infections. FIV enters the brain early following experimental infection, and virus has been proposed to enter the brain via the blood–brain barrier and blood–CSF barrier, within infected lymphocytes and monocytes/macrophages. However the entry of cell-free virus or the direct infection of brain endothelial cells and astrocytes of the blood–brain barrier may also contribute to CNS infection. This review explores the role played by the FIV model in the elucidation of mechanism of lentiviral entry to the brain and viral interactions with the CNS, particularly in relation to lymphotropic lentiviruses.     

A micro-enzyme-linked immunosorbent assay (ELISA) test for detection of feline leukemia virus in cats

The performance of a micro ELISA test for detection of feline leukemia virus (FeLV) infection was evaluated. The test was found specific for FeLV and feline sarcoma virus (FeSV) group-specific antigens in blood, plasma or serum of infected cats. Other common feline pathogens were negative to the test.Quantities as little as 7.8 ng of p-27 (the major group specific antigen of FeLV) per ml of sample gave positive results. The correlation between the micro ELISA test and the indirect immunofluorescent test commonly used for diagnosis of FeLV infection was 98% in 116 clinical cases and 184 samples from cats inoculated with FeLV and 100% in 100 specific pathogen-free cats.     

重组N蛋白抗原检测PRRSV抗体ELISA的研究Ⅳ.与IDEXX ELISA试剂盒及Western blot的比较

利用重组N蛋白抗原建立了检测猪繁殖与呼吸综合征病毒（PRRSV）抗体的ELISA方法并组装成试剂盒，将试制的3批试剂盒分别与IDEXX生产的试剂盒及Western—blot进行了符合率试验。试验结果表明，所研制的试剂盒与Western blot的符合率达97．67％以上。对460份临床血清分别用自制的试剂盒和IDEXX公司生产的试剂盒进行检测，其中有37份不相符合，用Western blot对这37份血清进行验证，有35份血清检测结果与自制的试刺盒检测结果一致。由此表明，自行研制的试剂盒，其特异性和敏感性均能满足目前临床上该疫病的流行病学分析或免疫抗体检测。