Ruiyu Pu

FIV vaccine development and its importance to veterinary and human medicine: a review FIV vaccine 2002 update and review.

Abstract Feline immunodeficiency virus (FIV) is a natural infection of domestic cats that results in acquired immunodeficiency syndrome resembling human immunodeficiency virus (HIV) infection in humans. The worldwide prevalence of FIV infection in domestic cats has been reported to range from 1 to 28%. Hence, an effective FIV vaccine will have an important impact on veterinary medicine in addition to being used as a small animal AIDS model for humans. Since the discovery of FIV reported in 1987, FIV vaccine research has pursued both molecular and conventional vaccine approaches toward the development of a commercial product. Published FIV vaccine trial results from 1998 to the present have been compiled to update the veterinary clinical and research communities on the immunologic and experimental efficacy status of these vaccines. A brief report is included on the outcome of the 10 years of collaborative work between industry and academia which led to recent USDA approval of the first animal lentivirus vaccine, the dual-subtype FIV vaccine. The immunogenicity and efficacy of the experimental prototype, dual-subtype FIV vaccine and the efficacy of the currently approved commercial, dual-subtype FIV vaccine (Fel-O-Vax FIV) are discussed. Potential cross-reactivity complications between commercial FIV diagnostic tests, Idexx Snap Combo Test® and Western blot assays, and sera from previously vaccinated cats are also discussed. Finally, recommendations are made for unbiased critical testing of new FIV vaccines, the currently USDA approved vaccine, and future vaccines in development.

Dual-subtype FIV vaccine protects cats against in vivo swarms of both homologous and heterologous subtype FIV isolates

ObjectiveTo evaluate the immunogenicity and efficacy of an inactivated dual-subtype feline immunodeficiency virus (FIV) vaccine. DesignSpecific-pathogen-free cats were immunized with dual-subtype (subtype A FIVPet and subtype D FIVShi) vaccine and challenged with either in vivo- or in vitro-derived FIV inocula. MethodsDual-subtype vaccinated, single-subtype vaccinated, and placebo-immunized cats were challenged with in vivo-derived heterologous subtype B FIVBang [10–100 50% cat infectious doses (CID50)], in vivo-derived homologous FIVShi(50 CID50), and in vitro- and in vivo-derived homologous FIVPet(20–50 CID50). Dual-subtype vaccine immunogenicity and efficacy were evaluated and compared to single-subtype strain vaccines. FIV infection was determined using virus isolation and proviral PCR of peripheral blood mononuclear cells and lymphoid tissues. ResultsFour out of five dual-subtype vaccinated cats were protected against low-dose FIVBang (10 CID50) and subsequently against in vivo-derived FIVPet (50 CID50) challenge, whereas all placebo-immunized cats became infected. Furthermore, dual-subtype vaccine protected two out of five cats against high-dose FIVBang challenge (100 CID50) which infected seven out of eight single-subtype vaccinated cats. All dual-subtype vaccinated cats were protected against in vivo-derived FIVPet, but only one out of five single-subtype vaccinated cats were protected against in vivo-derived FIVPet. Dual-subtype vaccination induced broad-spectrum virus-neutralizing antibodies and FIV-specific interferon-γ responses along with elevated FIV-specific perforin mRNA levels, suggesting an increase in cytotoxic cell activities. ConclusionDual-subtype vaccinated cats developed broad-spectrum humoral and cellular immunity which protected cats against in vivo-derived inocula of homologous and heterologous FIV subtypes. Thus, multi-subtype antigen vaccines may be an effective strategy against AIDS viruses.

Dual-subtype FIV vaccine (Fel-O-Vax® FIV) protection against a heterologous subtype B FIV isolate

Vaccine trials were undertaken to determine whether the Fel-O-Vax® FIV, a commercial dual-subtype (subtypes A and D) feline immunodeficiency virus (FIV) vaccine, is effective against a subtype B FIV isolate. Current results demonstrate the Fel-O-Vax FIV to be effective against a subtype B virus, a subtype reported to be the most common in the USA.

Efficacy evaluation of prime-boost protocol : canarypoxvirus-based feline immunodeficiency virus (FIV) vaccine and inactivated FIV-infected cell vaccine against heterologous FIV challenge in cats

Objective:To evaluate the immunogenicity and prophylactic efficacy of immunization schemes employing a recombinant canarypoxvirus (‘ALVAC’)-based feline immunodeficiency virus (FIV) vaccine alone or in combination with an inactivated FIV-infected cell vaccine against homologous and heterologous FIV challenges in cats. Methods:Specific pathogen-free cats were given a total of three immunizations with subtype A vaccines and challenged 4 weeks after the final immunization with 50 median animal infectious doses (ID50) of FIV-Petaluma, a subtype A isolate. Following the initial challenge, protected cats received a second challenge with 75 ID50 of FIV-Bangston, a subtype B isolate. FIV-specific humoral and cell-mediated responses were measured to determine the immune correlates of protection. Results:Two of three cats immunized with the ALVAC-FIV recombinants alone were protected from homologous FIV challenge in the presence of FIV-specific cytotoxic T-lymphocyte (CTL) responses but in the absence of FIV-specific humoral responses. All three cats immunized with the ALVAC-FIV recombinant and boosted with FIV-infected cell vaccine were also protected from homologous FIV challenge in the presence of both FIV-specific CTL and humoral responses. Partial to full protection was observed in ALVAC-FIV/FIV-infected cell vaccine-immunized cats against a heterologous FIV challenge given 8 months after the initial challenge. Two out of three cats had transient infection and the remaining cat had no sign of FIV infection at a dose at which all three control cats were readily infected. Conclusions:Immunization schemes employing ALVAC-based FIV vaccines in combination with inactivated FIV-infected cell vaccine generate protective immune responses that can cross-react with FIV isolates that are genetically distinct from the vaccine strains.

Protection of neonatal kittens against feline immunodeficiency virus infection with passive maternal antiviral antibodies

Objective: Maternal antibodies from either vaccinated or feline immunodeficiency virus (FIV)‐infected female cats (queens) were evaluated for their ability to protect kittens against homologous FIV infection. Design: Kittens that received different levels of maternal antiviral antibodies from either vaccinated or infected queens were inoculated with homologous FIV at 1 week post‐parturition and monitored for FIV infection. Maternal antiviral antibodies in the kittens were also measured and compared to the level of FIV infection. Methods: Kittens at 1 week post‐parturition were inoculated intraperitoneally with five median cat infectious doses of FIVPet. FIV infection was monitored by virus isolation for infectious FIV and by nested polymerase chain reaction for proviral DNA. Virus‐neutralizing (VN) antibodies and antibodies to FIV transmembrane peptide and core protein were also monitored throughout the 25 weeks. Results: Neonatal kittens that received high levels of antiviral antibodies from either vaccinated or infected queens were protected from FIV inoculation. Kittens that received low levels of maternal antiviral antibodies were not completely protected from similar FIV inoculation. Protection correlated more closely with the level of maternal VN antibodies than the anti‐p25 antibodies transferred to the kittens. The unprotected kittens born to infected queens were not infected from vertical transmission because all littermates that were not FIV‐inoculated remained free of FIV infection. Conclusions: Maternal antiviral antibodies, including VN antibodies, from either vaccinated or infected queens protected neonatal kittens from FIV inoculation. Thus, maternal antiviral antibodies play a key role in preventing or limiting infection in neonates and such antiviral immunity can be provided by vaccinated queens.

HIV-1 p24 vaccine protects cats against feline immunodeficiency virus infection.

Background:Based on previous analysis of feline immunodeficiency virus (FIV)-specific cross-reactive antibodies to HIV-1 p24, cats vaccinated with HIV-1 p24 were evaluated for cross-reactive immunity to FIV. Objective:To determine the level of cross-reactivity that exists between HIV-1 and FIV p24 and its implications for vaccine prophylaxis. Methods:Specific-pathogen-free cats were immunized three times with HIV-1 p24 in Ribi adjuvant, with (n = 18) or without cytokine (n = 6). Control cats were immunized three times with adjuvant (n = 10) or phosphate-buffered saline (PBS; n = 5). All immunized cats were challenged with either subtypes B or A/B FIV, and monitored by virus isolation, proviral PCR, FIV-specific antibodies, and feline interferon-γ ELISpot for T-cell activities. Results:Of 18 cats vaccinated with subtype B HIV-1 (HIV-1LAI/LAV, HIV-1UCD1) p24 in Ribi/cytokine adjuvant 14 (78%) were protected against FIV challenges (subtype Agag and Bgag) that infected all 15 adjuvant- or PBS-immunized cats. Furthermore, only three of six (50%) cats vaccinated with FIV p24 in Ribi/cytokine adjuvant were protected against similar FIV challenge. HIV-1 p24 vaccination induced weak cross-reactive antibodies to FIV p24, which did not correlate with vaccine efficacy. However, the peripheral blood mononuclear cells from HIV-1 p24-vaccinated/protected cats at 33–34 weeks post-FIV challenge responded to three T-cell responsive peptides at the carboxyl-terminus of the FIV p24, whereas those cells from the infected control cats had minimal to no responses to the same peptides. Conclusions:These results suggest the importance of including lentiviral p24 as vaccine immunogen for human AIDS vaccine. Moreover, these results suggest the potential importance of evolutionarily conserved, cross-protective epitopes in vaccine protection.

Development of FIV-specific cytolytic T-lymphocyte responses in cats upon immunisation with FIV vaccines.

Vaccine protection has been achieved in cats against experimental infection with feline immunodeficiency virus (FIV). Such protection has been attributed to FIV-specific humoral immunity, as well as cellular immunity of unknown mechanism(s). Since cell-mediated immunity plays a crucial role in the clearance of viral infections, this study evaluated the role of FIV-specific CTL in vaccine prophylaxis. Cats were immunised with inactivated FIV vaccines, reported to have > 90% vaccine efficacy. Significant levels of specific CTL activity were detected following the third immunisation. CTL activity persisted for several months and could be enhanced through a booster immunisation. The levels of CTL activity were comparable to those induced by a recombinant canarypoxvirus based FIV vaccine. These results suggest a possible role for CTL-mediated immunity in vaccine protection against FIV infection in cats.

FELINE BONE MARROW TRANSPLANTATION : ITS USE IN FIV-INFECTED CATS

The use of autologous and allogenic bone marrow transplantations (BMT) in FIV-infected and uninfected cats is a novel therapy for feline hematopoietic diseases and retroviral infections. A total of 13 specific pathogen-free (SPF) cats received either autologous or allogenic BMT and seven of these cats were also infected with FIV before autologous or allogenic BMT. All BMT recipients received total body irradiation of 900 cGy just before BMT. Two FIV-infected and four uninfected cats received autologous uninfected BM cells cryopreserved before BMT. Five infected and two uninfected cats received BM cells from allogenic uninfected donors (RBC-, MHC-, and cross-matched). MHC-matching was based on mixed leucocyte reaction (MLR) and the donor-recipient combination which was compatible by MLR analysis, was used in this study. Recipients were monitored for hematology, immunology, virology, and clinical signs. All FIV-infected and uninfected recipients of autologous BMT had complete engraftment with minimal complications. Uninfected recipients of allogenic BMT had a more severe clinical episode with slower rate of engraftment. None of these BMT groups had mortality. In contrast, only two of the five infected recipients of allogenic BMT survived for a significant period of time (23 and 50 weeks) and rest of the cats succumbed to transfusion reactions. Both infected BMT groups had persistent CD4/CD8 inversion, low CD4+ cell counts, and FIV infection of engrafted peripheral blood mononuclear cells (PBMC). Overall, successful autologous and allogenic BMTs were performed in FIV-free cats. All infected recipients of autologous BMT had compete engraftment and are currently alive, with thelongest survival time being over 1 year. Thus, BMT in combination with antiviral drug therapies may be an alternative therapy against retroviral infection.

Utilization of Feline ELISPOT for Mapping Vaccine Epitopes

A commercial feline immunodeficiency virus (FIV) vaccine consisting of inactivated dual-subtype viruses was released in the USA in 2002 and released subsequently over the next 6 years in Canada, Australia, New Zealand, and Japan. Based on the genetic, morphologic, and biochemical similarities between FIV and human immunodeficiency virus-1 (HIV-1), FIV infection of domestic cats is being used as a small animal model of HIV/AIDS vaccine. Studies on prototype and commercial FIV vaccines provide new insights to the types of immunity and the vaccine epitopes required for an effective human HIV-1 vaccine. ELISPOT assays to detect cytokines, chemokines, and cytolytic mediators are widely used to measure the magnitude and the types of cellular immunity produced by vaccination. Moreover, such approach has identified regions on both HIV-1 and FIV proteins that induce robust antiviral cellular immunity in infected hosts. Using the same strategy, cats immunized with prototype and commercial FIV vaccines are being analyzed by feline interferon-γ and IL-2 ELISPOT systems to identify the vaccine epitope repertoire for prophylaxis.

An initial examination of the potential role of T-cell immunity in protection against feline immunodeficiency virus (FIV) infection

The importance of vaccine-induced T-cell immunity in conferring protection with prototype and commercial FIV vaccines is still unclear. Current studies performed adoptive transfer of T cells from prototype FIV-vaccinated cats to partial-to-complete feline leukocyte antigen (FLA)-matched cats a day before either homologous FIVPet or heterologous-subtype pathogenic FIVFC1 challenge. Adoptive-transfer (A-T) conferred a protection rate of 87% (13 of 15, p < 0.001) against FIVPet using the FLA-matched T cells, whereas all 12 control cats were unprotected. Furthermore, A-T conferred protection rate of 50% (6 of 12, p<0.023) against FIVFC1 using FLA-matched T cells, whereas all 8 control cats were unprotected. Transfer of FLA-matched T and B cells demonstrated that T cells are needed to confer A-T protection. In addition, complete FLA-matching and addition of T-cell numbers > 13 × 10(6) cells were required for A-T protection against FIVFC1 strain, reported to be a highly pathogenic virus resistant to vaccine-induced neutralizing-antibodies. The addition of FLA-matched B cells alone was not protective. The poor quality of the anti-FIV T-cell immunity induced by the vaccine likely contributed to the lack of protection in an FLA-matched recipient against FIVFC1. The quality of the immune response was determined by the presence of high mRNA levels of cytolysin (perforin) and cytotoxins (granzymes A, B, and H) and T helper-1 cytokines (interferon-γ [IFNγ] and IL2). Increased cytokine, cytolysin and cytotoxin production was detected in the donors which conferred protection in A-T studies. In addition, the CD4(+) and CD8(+) T-cell proliferation and/or IFNγ responses to FIV p24 and reverse transcriptase increased with each year in cats receiving 1X-3X vaccine boosts over 4 years. These studies demonstrate that anti-FIV T-cell immunity induced by vaccination with a dual-subtype FIV vaccine is essential for prophylactic protection against AIDS lentiviruses such as FIV and potentially HIV-1.