Human immunodeficiency virus (HIV) is the retrovirus that causes acquired immunodeficiency syndrome (AIDS). Within bodily fluids, HIV is present as both free virus particles and as virus within infected immune cells. So far, AIDS has killed more than 25 million people since it was first recognized in 1981, making it one of the most destructive pandemics in recorded history. Antiretroviral treatment reduces both the mortality and the morbidity of HIV infection, but routine access to antiretroviral medication is not available in all countries. Most HIV-infected individuals develop AIDS and die; however, about 10% remains healthy for many years, with no noticeable symptoms. Treatment with anti-retrovirals increases the life expectancy of people infected with HIV (Palella et al., 1998; Buchbinder et al., 1994).
4. Microbial Pathogenesis
The 3 major routes of HIV transmission are sexual intercourse, contaminated needles, and transmission from an infected mother to her baby at birth, or through breast milk. HIV primarily infects vital cells in the human immune system such as helper T cells (specifically CD4+ T cells), macrophages, and dendritic cells. This decreases CD4+ T cells through direct viral killing of infected cells, increased rates of apoptosis in infected cells, and killing of infected CD4+ T cells by CD8 cytotoxic lymphocytes that recognize infected cells. When CD4+ T cell numbers decline below a critical level, cell-mediated immunity (CMI) is lost, and the body becomes progressively more susceptible to opportunistic infections (Chan et al., 1997).
5. Host Ranges and Animal Models
AIDS has killed more than 25 million people since it was first recognized in 1981. Globally, ~40 million people currently live with HIV. Sub-Saharan Africa remains by far the worst-affected region. More than 64% of all people living with HIV are in sub-Saharan Africa, as are more than three quarters of all women living with HIV. Two-thirds of HIV/AIDS infections in Asia occur in India, with an estimated 6 million infections, surpassing South Africa's 5.5 million infections, making India the country with the highest number of HIV infections in the world (Palella et al., 1998).
6. Host Protective Immunity
HIV can infect a variety of immune cells such as CD4+ T cells, macrophages, and microglial cells. HIV-1 entry to macrophages and T cells is mediated through interaction of the virion envelope glycoproteins (gp120) with the CD4 molecule on target cells and also with chemokine coreceptors. Macrophages play a key role in several critical aspects of HIV infection. They appear to be the first cells infected by HIV and perhaps the source of HIV production when CD4+ cells become depleted in the patient. T-tropic isolates, or syncitia-inducing (SI) strains replicate in primary CD4+ T cells as well as in macrophages. Dual-tropic HIV-1 strains are thought to be transitional strains of the HIV-1 virus. HIV can also infect a subtype of myeloid dendritic cells, which probably constitute a reservoir that maintains infection when CD4+ T cell numbers have declined to extremely low levels (Chan et al., 1997; Coakley et al., 2005; Deng et al., 1996; Knight et al., 1990).
II. Vaccine Related Pathogen Genes
1. env
Gene Name :
env
Sequence Strain (Species/Organism) :
Human immunodeficiency virus 1
Protein Note :
isolated from patient SC49 PBMC; infection was aquired in Sydney, Australia 1998; this patient registered low levels of viral RNA in plasma by RT-PCR (Amplicor 1.0) and low levels of P24 (gag) antigen by monoclonal ELISA despite symptomatic seroconversion; HIV RNA quantitation by an alternative method (bDNA, Chiron 3.0) on the same sample yielded a level of >500,000 copies/ml; subtyping in envelope and P17 gag by sequencing and phylogenetic analysis revealed subtype B; the entire gag gene was then sequenced to search for mutations to account for the failure of detection methods directed to this genomic area; subtype: B
Molecule Role Annotation :
HIV-1 DNA plasmids expressing Env, Gag and Tat, either alone or combined with the corresponding protein immunogens formulated in the MF59 adjuvant, resulted in protection against an experimental HIV-1 infection in C57Bl/6 mice (Bråve et al., 2007).
Molecule Role Annotation :
A novel primate lentivirus was identified in Mandrillus sphinx. One of the newly isolated SIV strains was intravenously inoculated into two rhesus macaques and resulted in chronic infection. These SIVs do not cluster with SIVmndGB1, the former representative strain of SIVmnd, but form a new, independent lineage. These results indicate that these SIVmnds are a novel type of SIVmnd and have infected Cameroonian mandrills for a long time. To date, M. sphinx is the only primate species other than humans that is naturally infected with two different types of SIV (Takehisa et al., 2001).
Protein Note :
isolated from a 37 year old injecting drug user; HIV-1 RNA was extracted from plasma; RNA was reverse transcribed and the gp120 region was amplified using nested PCR; sequence was obtained bi-directionally from cloned DNA
Protein Note :
heptad repeat 1-heptad repeat 2 region (ectodomain) of the gp41 subunit of human immunodeficiency virus (HIV-1), and related domains; cd09909
Protein Note :
This particular nucleotide sequence has a premature stop codon in place of a well-conserved tryptophan codon at position 8712-8714 that truncates the HIV1 Nef protein sequence to a 123 amino acids-long N-terminal portion (not shown).
Molecule Role Annotation :
Study compared the handling of overlapping 24-mer peptides from HIV Nef either mixed or covalently linked in tandem in one protein. With adjuvant, overlapping peptides were capable of protecting Balb/c mice against lethal viral challenge with 3 × 108 plaque forming units of vv-nef, whereas the intact protein was less protective (Zhang et al., 2009).
>NP_057853.1 Tat [Human immunodeficiency virus 1]
MEPVDPRLEPWKHPGSQPKTACTNCYCKKCCFHCQVCFITKALGISYGRKKRRQRRRAHQNSQTHQASLS
KQPTSQPRGDPTGPKE
Molecule Role :
Protective antigen
Molecule Role Annotation :
HIV-1 DNA plasmids expressing Env, Gag and Tat, either alone or combined with the corresponding protein immunogens formulated in the MF59 adjuvant, resulted in protection against an experimental HIV-1 infection in C57Bl/6 mice (Bråve et al., 2007).
>AAB47786.1 tat protein [Human immunodeficiency virus 2]
METPLKAPESSLKSYNEPSPCTSEWEVAAQELAKQGEELLAQLHRPLEACTNPCYCKKCSFHCQLCFLKK
GLGIWYVRKGGRRRTPKRTKTHPPSTPDKSISIQTGDSRPTKKQKETPETPVATTSGLGR
Protein Note :
An artificial frameshift eliminating the orf-disrupting nucleotide at position 5320 is introduced to obtain the typical HIV-1 Vpr protein sequence. For this particular HIV-1 strain, HXB2, only a short (78 amino acid long) variant of the Vpr sequence can be obtained by translation of nucleotides 5105 through 5341 without the frameshift
Molecule Role Annotation :
IFN-gamma plays a critical role in Th1 type immune response. It is important for protection against infections by various viruses and intracellular bacteria.
Additional Molecule Role :
Vaximmutor
Additional Molecule Role Annotation :
The experimental data demonstrated that three time vaccinations with BCG in BALB/c mice induced strong TB Ag-specific IFN-gamma immune responses in splenocytes (Wang et al., 2009).
The adenoviral vector was based on a serotype 5 adenovirus that is incompetent to replicate with deletion of the E1 and E3 viral genes, and was propagated subsequently in E1-expressing 293 cells. Recombinant adenovirus expressing the codon-optimized SIV gag gene was then prepared. The recombinant adenovirus (Ad5-SIVgag) was grown in large quantities by multiple rounds of amplification in 293 cells. The virus was purified by caesium chloride gradient centrifugation (Shiver et al., 2002).
h. Immunization Route
Intramuscular injection (i.m.)
i.
Monkey Response
Host Strain:
rhesus monkey (Macaca mulatta)
Vaccination Protocol:
In this study, 21 rhesus monkeys were divided into 6 groups, including an unimmunized control cohort. Each of the test vectors expressed the identical SIVmac239 gag gene that had been codon optimized for expression in mammalian cells. In the first study, the two viral vector vaccines were administered, followed by a booster dose; preparations of DNA plasmid vector vaccine were delivered thrice, followed by a booster (Shiver et al., 2002).
Immune Response:
In general, all of the monkeys developed p11CM-specific cellular immune responses after the initial immunization series. The p11CM (residues 181–189) is an immunodominant SIV gag epitope that is presented by the Mamu-A*01 MHC protein and is capable of binding T-cell receptors in the model monkeys. Administration of the third dose of the Ad5 vector resulted in an additional increase of p11CM-specific CD8+ T cells at the time of virus challenge. After the booster inoculation, these animals exhibited peak levels of p11CM-specific CD8+ T cells (Shiver et al., 2002).
Challenge Protocol:
At 12 weeks after the final immunization, all monkeys were challenged i.v. with the pathogenic HIV–SIV hybrid virus (SHIV) 89.6P16. The challenge of the control and immunized animals within the context of each of the two independent studies occurred concurrently (Shiver et al., 2002).
Efficacy:
Each of the animals in both control groups exhibited acute CD4+ T-cell lymphopenia and peak viral loads of viral RNA copies at about 3 weeks after challenge. With the exception of one animal, all of the control monkeys experienced dramatic loss of CD4+ T cells. During the acute phase of the infection, most monkeys immunized with either the DNA or MVA vectors exhibit an acute CD4+ T-cell lymphopenia. By about 70 d after challenge, many of the immunized monkeys exhibit some evidence of a positive immunization benefit, as manifested by control of viremia and recovery of CD4+ T-cell counts. However, the animals immunized with Ad5 vector exhibited the most pronounced attenuation of the infection with a pathogenic HIV–SIV hybrid virus (SHIV) (Shiver et al., 2002).
Description:
Live non-pathogenic HIV-2 following priming with ALVAC HIV-2 (recombinant canarypox virus expressing HIV-2 env, gag and pol) (Walther-Jallow et al., 2001).
Description:
Live non-pathogenic HIV-2 following priming with ALVAC HIV-2 (recombinant canarypox virus expressing HIV-2 env, gag and pol) (Walther-Jallow et al., 2001).
A live non-pathogenic HIV-2 following priming with ALVAC HIV-2 (recombinant canarypox virus expressing HIV-2 env, gag and pol) (Walther-Jallow et al., 2001).
h. Immunization Route
Intramuscular injection (i.m.)
i.
Monkey Response
Vaccination Protocol:
Five monkeys were immunized either three times with ALVAC HIV-2 alone or twice with ALVAC HIV-2 and once with purified native HIV-2 gp125 (Walther-Jallow et al., 2001).
Vaccine Immune Response Type:
VO_0003057
Challenge Protocol:
The monkeys were then challenged with HIV-2 given intravenously and finally with pathogenic SIVsm given intrarectally (Walther-Jallow et al., 2001).
Efficacy:
After challenge with SIVsm, three of five monkeys were completely protected against SIVsm infection. Vaccination with an ALVAC HIV-2 vaccine followed by exposure to live HIV-2 could induce cross-protection against mucosal infection with SIVsm and seemed to be more efficient than immunization with a live HIV-2 vaccine only (Walther-Jallow et al., 2001).
Description:
Live attenuated human immunodeficiency virus type 2 (HIV-2) vaccine alone versus boosting with live non-pathogenic HIV-2 following priming with ALVAC HIV-2 (recombinant canarypox virus expressing HIV-2 env, gag and pol) (Walther-Jallow et al., 2001).
Description:
Live attenuated human immunodeficiency virus type 2 (HIV-2) vaccine alone versus boosting with live non-pathogenic HIV-2 following priming with ALVAC HIV-2 (recombinant canarypox virus expressing HIV-2 env, gag and pol) (Walther-Jallow et al., 2001).
A live non-pathogenic HIV-2 following priming with ALVAC HIV-2 (recombinant canarypox virus expressing HIV-2 env, gag and pol) (Walther-Jallow et al., 2001).
h. Immunization Route
Intramuscular injection (i.m.)
i.
Monkey Response
Vaccination Protocol:
Six monkeys were first inoculated intravenously with live HIV-2(SBL-6669) (Walther-Jallow et al., 2001).
Vaccine Immune Response Type:
VO_0003057
Challenge Protocol:
7 to 10 months after vaccination, the monkeys were challenged intrarectally with 10 MID(50) of cell-free simian immunodeficiency virus (SIV) strain SIVsm (Walther-Jallow et al., 2001).
Efficacy:
Vaccination with an ALVAC HIV-2 vaccine followed by exposure to live HIV-2 could induce cross-protection against mucosal infection with SIVsm and seemed to be more efficient than immunization with a live HIV-2 vaccine only (Walther-Jallow et al., 2001).
A live coxsackievirus B4 recombinant, CVB4/p24(73(3)), that expresses seventy-three amino acids of the gag p24 sequence (HXB2) (Gu et al., 2010).
f. Immunization Route
Intramuscular injection (i.m.)
g.
Mouse Response
Vaccination Protocol:
Immunization was by intraperitoneal (IP) injection or by oral gavage. Mice were immunized with either CVB4/p24(733) or the parental CVB4. PBS-treated mice served as controls (Gu et al., 2010).
Vaccine Immune Response Type:
VO_0003057
Challenge Protocol:
Mice immunized with the avirulent CVB4 variant are protected when subsequently challenged with a virulent variant (Gu et al., 2010).
Efficacy:
Results showed that oral immunization with CVB4/p24(73(3)) induced gag p24-specific immune responses in vector-immune mice (Gu et al., 2010).
The recombinant vaccinia viruses (rVac) expressing SHIV89.6P env and SIVmac239 gag were constructed by inserting these genes in the HindIII M region of TBC-Wy, Therion strain of vaccinia. rFPV viruses expressing these same genes were constructed by inserting the genes in the BamJHI region of POXVAC-TC (Schering-Plough) strain of FPV (Santra et al., 2004).
h. Description
An HIV vaccine should elicit a cytotoxic T lymphocyte (CTL) response, but the characteristics of effective vaccine-induced CTL response remain unclear. The SHIV/rhesus monkey model has been used to in the course of assessing the relative immunogenicity of vaccine regimens that include a cytokine-augmented plasmid DNA prime and a boost with DNA or recombinant pox vectors. This study indicates that the steady-state memory, rather than the peak effector vaccine-elicited CTL responses, may be the critical immune correlate of protection for a CTL-based HIV vaccine (Santra et al., 2004).
i.
Monkey Response
Host Strain:
rhesus monkey (Macaca mulatta)
Vaccination Protocol:
Nonrecombinant wildtype vaccinia virus was designated VV-WT, wild-type fowlpox virus was designated FPV-WT, and wild-type MVA was designated MVA-WT. These wild-type viruses were used as control vector immunogens (Santra et al., 2004). One group of monkeys were vaccinated by separate i.m. injections of HIV-1. Half the dose was delivered to each quadriceps muscle. Seven of the 28 monkeys were vaccinated by both i.d. and i.m. injections of rFPV expressing HIV-1 89.6P Env and the same virus expressing SIV mac239 Gag. Seven monkeys received rMVA-HIV-189.6P Env and rMVA-SIVmac239 Gag, and another seven monkeys received rVac-HIV-1 89.6P Env and rVac-SIVmac239 Gag administered both i.d. and i.m. Another 28 monkeys received sham plasmid DNA and empty pox vectors (Santra et al., 2004).
Persistence:
Persistence levels were measured 2 weeks after challenge with results mentioned in Challenge Protocol section (Santra et al., 2004).
Immune Response:
Recombinant vaccinia virus, MVA, and fowlpox were comparable in their immunogenicity. Magnitude of peak vaccine-elicited CTL responses in pox virus-boosted monkeys is substantially greater than that seen in monkeys immunized with plasmid DNA alone, but magnitudes of recombinant pox boosted CTL responses decayed rapidly and were comparable to those of the DNA-alone vaccinated monkeys by the time of viral challenge. Clinical protection seen in all groups of experimentally vaccinated monkeys is similar, indicating that steady-state memory, rather than peak effector vaccine-elicited T lymphocyte responses, may be the critical immune correlate of protection for a CTL-based HIV vaccine (Santra et al., 2004).
Challenge Protocol:
Eighteen weeks after the final immunization, all animals were challenged with cell-free SHIV-89.6P i.v. (Santra et al., 2004).
Efficacy:
Control monkeys developed tetramer-binding CD8 T lymphocyte responses that were maximal 2 weeks after viral challenge and no detectable p41A-specific CD8 T cells. In contrast, all four groups of vaccinated monkeys developed robust secondary p11C-specific CTL responses that were comparable in magnitude. Animals boosted with plasmid DNA had Gag p11C-specific CD8 T cell responses similar in magnitude to that seen in the recombinant pox virus-boosted animals. Magnitudes of the postchallenge IFN ELISPOT responses to both vaccine antigens were comparable in all four experimentally vaccinated groups of monkeys. Therefore, although the pre-challenge peak vaccine-elicited immune responses were greater in the groups of monkeys boosted with recombinant pox vectors, the pre-challenge plateau and post-challenge peak secondary responses were equivalent in magnitude in all four experimental groups of animals (Santra et al., 2004).
Host Gene Response of
IFNG
Gene Response:
There was a greater contraction of the vaccine-elicited IFN-γ-secreting T cell responses in the groups of monkeys boosted with recombinant pox vectors than in the group of animals boosted with plasmid DNA. Although the pre-challenge peak vaccine-elicited immune responses were greater in the groups of monkeys boosted with recombinant pox vectors, the pre-challenge plateau and post-challenge peak secondary responses were equivalent in magnitude in all four experimental groups of animals (Santra et al., 2004).
The antigens are named MultiHIV-A (based on the subtype A consensus sequence), MultiHIV-B (subtype B consensus sequence), MultiHIV-C, and MultiHIV-FGH (based on ancestral sequence for subtypes F, G, and H). The MultiHIV DNAs encode polypeptides consisting of a fusion of the full-length regulatory proteins Rev, Nef and Tat as well structural proteins p17 and p24 (Malm et al., 2005).
A vaccine platform was constructed with an HIV-1 subtype B DNA immunogen expressing full-length consensus sequences from HIV-1 rev, nef, tat, and gag with additional cellular epitope clusters from the env and pol regions. Furthermore, this platform has been extended to three additional plasmids expressing the same immunogens but originating from subtypes A or C consensus or FGH ancestral sequences (Malm et al., 2005).
l. Description
A significant limitation for HIV vaccine development is that there are no small animals in which actual productive HIV-1 infection can be established. DNA immunization with candidate vaccines comprising multiple genes of clades A, B, C, F, G, and H create strong cellular responses in BALB/c mice, especially after gene gun immunization (Malm et al., 2005).
m.
Mouse Response
Host Strain:
C57BL/6, BALB/c
Vaccination Protocol:
C57BL/6 mice were divided into groups g.g. immunized or i.m. inoculated. Mice were bled 2 weeks after the last immunization, and individual blood and spleen samples were collected post-challenge and used fresh in ELISPOT assays (Malm et al., 2005).
BALB/c were immunized by MultiHIV/MultiClade DNA. Initially the immune response was evaluated after 3 immunizations by g.g. Equal amounts of clade A, B, C, and FGH MultiHIV plasmids were mixed together and coated onto the gold particles to construct the subtype cocktail MultiHIV DNA immunogen (MultiHIVmix). Mice were sacrificed 10 days after the last immunization and individual spleens were collected and the cells preserved at –70°C until used. In a second set of experiments a short-term immunization schedule was used to address three different administration routes; g.g., i.m., and i.d. Mice were immunized 3 times with MultiHIVmix DNA (Malm et al., 2005).
Immune Response:
Though the immune response detected was quite low, the majority of g.g. immunized animals responded with specific IFN production. Interestingly, the gag specific response in the same gene gun immunized animals was weaker than the envelope responses and also drastically weaker than the responses seen before the experimental challenge (Malm et al., 2005). The CTL response, following i.d. injection, was detected five weeks after the first immunization and persisted up to 12 weeks later. In contrast, i.m. immunization induced detectable IFNy only at week 17 (Malm et al., 2005).
Challenge Protocol:
C57BL/6 mice transgenic for HLA-A201 were given an experimental challenge after a short-term immunization schedule. Amphotropic murine leukemia virus was used to prepare pseudovirus with HIV-1 isolate. Challenged animals were given i.p. injections of sHIV-1/MuLV infected cells (Malm et al., 2005).
Efficacy:
The vaccine-induced immunity has in vivo efficacy. Here a mouse model showed that the HIV-1 GTU-MultiHIVmix vaccine induces virus-specific CD8 T cellular immunity and protects against experimental HIV-1 challenge (Malm et al., 2005).
Description:
The cocktail of MultiHIV protected 19/24 animals against an experimental HIV-1 challenge. This experiment also demonstrates cross-clade protection, as the subtypes A and B viruses used for challenge were derived from different subtypes than the sequences found in the clade A or B specific MultiHIV constructs. Furthermore, it demonstrates that consensus approach used in Multi-HIVmix vaccine is functional and causes protection against naturally occurring isolates. Gene gun immunization was superior to intramuscular immunization in terms of both the T cell immunity induced, as well as higher frequency of protection. Data generated in this work support the hypothesis that DNA representing several HIV-1 subtypes and several genes are immunogenic and protective (Malm et al., 2005).
Host Gene Response of
Ifng (Interferon gamma)
Gene Response:
IFN-gamma levels in mice immunized by gene gun or intramuscular delivery with HIV-1 subtype B DNA immunogen showed strong up regulation in response to an H-2d restricted gag peptide (AMQMLKETI) and even stronger up-regulation in response to an env epitope (RGPGRAFVTI) (Malm et al., 2005).
HIV matrix-capsid portion of Gag, envelope gp160, secreted gp140, cloned SIVsm H-4i, SIVsm E660 (Davis et al., 2002)
c. Vector:
VEE replicon particles (VRPs)
d. Preparation
Gag-VRPs is a cocktail vaccine of V3014-packaged VRPs expressing the SIVsm H-4i nonmyristylated matrix-capsid region, full-length gp160, and a secreted form of gp160 (gp140). Structural proteins for packaging of replicon RNA into VRPs are expressed from separate helper RNAs. VRPs expressing either the matrix-capsid portion of Gag, the full-length envelope gp160, or the secreted gp140 of cloned SIVsm H-4i were mixed in a cocktail and used to immunize macaques (Davis et al., 2002).
e.
Mouse Response
Host Strain:
BALB/c
Vaccination Protocol:
VRPs with wild-type glycoprotein spikes were inoculated into the footpads of mice (Davis et al., 2002).
Persistence:
At 11 months post-boosting with the downstream vector, serum antibody levels against HIV MA/CA were undiminished, and MA/CA specific CTLp were detectable in all mice tested. These findings suggest that VEE vectors can be optimized to elicit strong, balanced and long-lived immune responses to foreign viral proteins (Caley et al., 1999).
Immune Response:
In BALB/c mice, the two vectors elicited cellular immune responses to MA/CA as determined by bulk CTL assays and precursor frequency analysis, but the humoral response induced by the downstream vector was significantly stronger. These findings suggest that VEE vectors can be optimized to elicit strong, balanced and long-lived immune responses to foreign viral proteins (Caley et al., 1999).
f.
Monkey Response
Host Strain:
rhesus monkey (Macaca mulatta)
Vaccination Protocol:
A cocktail vaccine of V3014-packaged VRPs expressing the SIVsm H-4i nonmyristylated matrix-capsid region, full-length gp160, and a secreted form of gp160 (gp140) was used to immunize rhesus macaques. Animals were given each VRP subcutaneously in the arm and later were challenged by the intrarectal (IR) route. Control animals received an equivalent dose of HA-VRPs (Davis et al., 2002).
Immune Response:
Both humoral and cellular immune responses were induced. On the day of challenge, all vaccinated animals had neutralizing antibody to the homologous SIVsm H-4, most had CTL specific for Gag, Env, or both. The animals were followed for a period of 40 weeks postchallenge, and although vaccination did not prevent infection by the high dose IR challenge, several protective effects of vaccination were seen. Peak virus titers in plasma were reduced, and the range of peak titers was much smaller for the controls, suggesting that a clear protective effect against the acute phase of infection was induced in some of the vaccinated animals (Davis et al., 2002).
Challenge Protocol:
Animals were given a dose of VRP subcutaneously in the arm and 1 month later were challenged by the intrarectal (IR) route. Control animals received an equivalent dose of HA-VRPs. The challenge virus was the highly virulent swarm SIVsm E660 (Davis et al., 2002).
Efficacy:
Animals were followed for a period of 40 weeks postchallenge, and although vaccination did not prevent infection by the high dose IR challenge, several protective effects of vaccination were seen. Four of six vaccinated animals, as compared to one of six controls, showed virus loads below 1,700 copies per ml at the “set point” (23 weeks postchallenge). By 41 weeks postchallenge, when the experiment was terminated, there was a significant decrease in the mean plasma virus load in the vaccinated animals compared to that in the controls. Most vaccinated animals showed virus loads below the “set point” (23 weeks postchallenge). By 41 weeks postchallenge, when the experiment was terminated, there was a significant decrease in the mean plasma virus load in the vaccinated animals compared to that in the controls. Finally, the CD4C cells of the vaccinated animals were preserved and even increased postchallenge compared to those of the controls. In fact, in the vaccinated animals, there is a clear correlation between increased CD4C cells and lowered viral load (Davis et al., 2002).
Vaccination Protocol:
Mice were subcutaneously immunized with antigen and adjuvant (Bomford et al., 1992).
Immune Response:
Mice immunized with gp120 and GMDP in combination with pluronic emulsion resulted in a secondary antibody response (Bomford et al., 1992).
Vaccination Protocol:
Mice were subcutaneously immunized with antigen and adjuvant (Bomford et al., 1992).
Immune Response:
Mice immunized with gp120 adjuvanted with MPD plus pleuronic emulsion resulted in a secondary antibody response (Bomford et al., 1992).
Vaccination Protocol:
The protein vaccine component consisted of 20 μg ro-gp140 delivered in the right tibialis muscle and 20 μg rGag and 5 μg rTat delivered in the left tibialis muscle. The proteins were formulated in 50% MF59 solution. Mock treatment consisted of MF59 adjuvant alone or empty DNA plasmid alone (Bråve et al., 2007).
Challenge Protocol:
Briefly, 3 weeks after the final immunization (week 12), the animals were injected intraperitoneally with 1 million syngeneic splenocytes infected with HIV-1/MuLV of subtype B (LAI) (Bråve et al., 2007).
Efficacy:
83% of mice were tested negative for HIV-1 after challenge (Bråve et al., 2007).
12. HIV DNA and adenoviral vector Ad5 expressing SIV gag protein
The V1R DNA vector expressed the identical SIVmac239 gag gene that had been codon optimized for expression in mammalian cells (Shiver et al., 2002). The plasmid DNA vector was formulated in a solution containing a nonionic blocked copolymer adjuvant (CRL1005) (Shiver et al., 2002). The adenoviral vector was based on a serotype 5 adenovirus that is incompetent to replicate with deletion of the E1 and E3 viral genes, and was propagated subsequently in E1-expressing 293 cells. Recombinant adenovirus expressing the codon-optimized SIV gag gene was then prepared. The recombinant adenovirus (Ad5-SIVgag) was grown in large quantities by multiple rounds of amplification in 293 cells. The virus was purified by caesium chloride gradient centrifugation (Shiver et al., 2002).
j. Immunization Route
Intramuscular injection (i.m.)
k. Virulence
No virulence has been reported associated with this vaccine (Shiver et al., 2002).
l. Description
Immunization and viral challenge studies were conducted in rhesus macaques (Macaca mulatta). Each of the test vectors expressed the identical SIVmac239 gag gene that had been codon optimized for expression in mammalian cells. The plasmid DNA vector was formulated in either PBS solution, a solution containing adjuvant, or a solution containing MPL/alum. All immunized animals were genotyped for the MHC class I Mamu-A*01 allele, allowing analysis of the responses of T cells (CD8+) to the test vaccines (Shiver et al., 2002).
m.
Monkey Response
Host Strain:
rhesus monkey (Macaca mulatta)
Vaccination Protocol:
This study used 14 monkeys, 3 each in the immunized groups and 8 in the unimmunized control group. Each of the test vectors expressed the identical SIVmac239 gag gene that had been codon optimized for expression in mammalian cells. The DNA vector priming inoculations were administered, followed by viral vector boost inoculations (Shiver et al., 2002).
Immune Response:
In general, all of the monkeys developed p11CM-specific cellular immune responses after the initial immunization series. The p11CM (residues 181–189) is an immunodominant SIV gag epitope that is presented by the Mamu-A*01 MHC protein and is capable of binding T-cell receptors in the model monkeys. Administration of the third dose of the Ad5 vector resulted in an additional increase of p11CM-specific CD8+ T cells at the time of virus challenge. After the booster inoculation, these animals exhibited peak levels of p11CM-specific CD8+ T cells (Shiver et al., 2002).
Side Effects:
None of the vaccinated monkeys have yet exhibited any signs of immunodeficiency or suffered any consistent weight loss (Shiver et al., 2002).
Challenge Protocol:
At 6 weeks after the final immunization, all monkeys were challenged i.v. with the pathogenic HIV–SIV hybrid virus (SHIV) 89.6P16. The challenge of the control and immunized animals within the context of each of the two independent studies occurred concurrently (Shiver et al., 2002).
Efficacy:
Each of the animals in both control groups exhibited acute CD4+ T-cell lymphopenia and peak viral loads of viral RNA copies at about 3 weeks after challenge. All of the control monkeys experienced dramatic loss of CD4+ T cells. During the acute phase of the infection, most monkeys immunized with either the DNA or MVA vectors or with the DNA/CRL1005–MVA vector prime–boost combination exhibit an acute CD4+ T-cell lymphopenia. By about 70 d after challenge, many of the immunized monkeys exhibit some evidence of a positive immunization benefit, as manifested by control of viremia and recovery of CD4+ T-cell counts. However, the animals immunized with Ad5 vector exhibited the most pronounced attenuation of the infection with a pathogenic HIV–SIV hybrid virus (SHIV) (Shiver et al., 2002).
Host Gene Response of
IFNG
Gene Response:
Levels of interferon-gamma in monkeys stimulated intramuscularly with either the p11CM peptide or a pool of peptides derived from SIV gag were significantly upregulated and correlated well with the tetramer-staining results after the final boost. Intracellular IFN-[gamma]-staining assays confirmed that these responses were primarily mediated by CD8+ T cells (Shiver et al., 2002).
Efficacy:
After the high-dose vaginal SHIV-162P3 challenge, 7/8 control animals were infected. Similarly, 7/8 animals in the vaccine-only group (group V) were infected. In the microbicide-only group (group M), 4/8 animals were infected. In the combination group (group V+M), only 2/7 animals were infected. The intervention efficacy in group V+M was 67%. In summary, the data supports that vaccines and microbicides used in combination may confer protection against sexual HIV-1 transmission to women by the vaginal route (Barouch et al., 2012).
Efficacy:
Animals were challenged with a heterologous chimpanzee titered stock of HIV-1 SF2 virus. Polymerase chain reaction coupled with reverse transcription results indicated infection in the control animal, whereas those animals vaccinated with the DNA constructs were protected from the establishment of infection (Boyer et al., 1997).
full-length unmutated SIVmac239 Gag and Pol, HIV-1AD8, Tat, Rev, and Vpu, and the 5′ third of HIV-1AD8 Env (Dale et al., 2004)
f. Gene Engineering of
env
Type:
DNA vaccine construction
Description:
The DNA vaccine strain, pHIS-SHIV-B, encoded full-length unmutated SIVmac239 Gag and Pol, HIV-1AD8, Tat, Rev, and Vpu, and the 5′ third of HIV-1AD8 Env (Dale et al., 2004).
Efficacy:
DNA vaccination alone primed for protection almost as effectively as the DNA/fowlpox virus regimen (Dale et al., 2004).
Host Gene Response of
IFNG from Macaca nemestrina
Gene Response:
SIV Gag-specific CD8 T cells expressing IFN-γ were dramatically boosted after challenge. Mean Gag-specific CD8 T cells peaked 3 weeks following challenge at 11.7% of all CD8 T cells in these outbred pigtail macaques immunized with the 2DNA/FPV regimen. The other vaccine groups had lower mean postchallenge CD8 T-cell responses, with peak responses at week 2 following challenge numbering 3.8% in the 3DNA group, 1.1% responses in the 1DNA/FPV group, and 2.0% in the 2DNA/FPV-IFN-γ-immunized animals (Dale et al., 2004).
Immune Response:
Co-delivery of GM-CSF and vaccine DNAs enhanced the temporal appearance of neutralizing Ab and broadened the specificity of the neutralizing activity to include SHIV-89.6P (Robinson et al., 2006).
Efficacy:
The GM-CSF-adjuvanted group showed a trend towards better control of the challenge infection and had better control of re-emergent virus (P < 0.01) than the non-adjuvanted group. After 52 weeks, the non-GM-CSF group continued to show some peaks of re-emergent virus whereas the GM-CSF group continued to maintain control below our level of detection (Robinson et al., 2006).
17. HIV DNA vaccine SIVmac239 Gag-Pol-Nef and mock Env with rAd5 boost
Immune Response:
DNA priming-rAd5 boosting immunization platform can elicit potent anti-HIV-1 cellular immune responses and anti-HIV-1 neutralizing antibodies (Mascola et al., 2005).
Efficacy:
In a challenge study, all monkeys were immunized with DNA plasmids at 0, 4, and 8 weeks and boosted with rAd5 at week 26. Serial plasma dilutions were tested against four of the more neutralization-sensitive viruses, and the plasma dilution that produced 50% virus neutralization, which is required to protect 50% of the cells from virus-induced killing, was determined. The data clearly demonstrates that both the vaccine platform and these immunogens elicit IgG-mediated virus neutralization (Mascola et al., 2005).
Immune Response:
The secretion of IFN-y stimulated CD4+ Th cell lines and rgp120-stimulated PBL from the vaccinated monkeys suggest that an HIV-1 env plasmid DNA vaccine elicits a Thl-like immune response in primates as well as in rodents (Lekutis et al., 1997)
Efficacy:
The secondary immune response that arose after repeated plasmid DNA administration was Thl-like, suggesting that the nature of the DNA vaccine-elicited Th cell response was maturation dependent. Evidence from vaccinated nonhuman primates suggests that a vigorous Th cell response, including viral Ag-specific IFN-y production, may be a correlate of protection from HIV- 1 and Simian Human Immunodeficiency Virus infection (Lekutis et al., 1997)
19. HIV priming with DNA vaccine expressing HIV gp160 protein and boosted with Ad5/35 vector expressing the same protein
pCAGGS and a replication-defective chimeric Ad5 vector with the Ad35 fiber (Ad5/35)
h. Preparation
A replication-defective chimeric Ad5 vector with the Ad35 fiber (Ad5/35) was prepared and used to express HIV Env gp160 protein. The product is named Ad5/35-HIV (Xin et al., 2005).
i. Virulence
This novel Ad5/35 vector showed minimal hepatotoxicity after intramuscular administration with the novel Ad5/35 vector (Xin et al., 2005).
j. Description
Replication-defective Ad5 HIV recombinants and replication-defective MVA elicit potent CD8+ T-cell responses and provide a high degree of protection in NHPs. The Ad5 (subgroup C) has well-defined biological properties and has been widely used as a vector for gene therapy and vaccine. The replication-defective Ad5 vector can easily be produced in high titers and is highly effective in boosting HIV-specific immunity. However, this virus uses CAR as its primary attachment receptor, which confers tropism for liver parenchymal cells. This raises important safety concerns. Thus, a replication-defective chimeric Ad5 vector with Ad type 35 fiber (Ad5/35) has been developed, which not only induces strong antigen-specific humoral and cellular immune responses and exhibits minimal hepatotoxicity in both mice and NHPs, but is also significantly less susceptible to the pre-existing Ad5 immunity than a comparable Ad5 vector (Xin et al., 2005).
k.
Monkey Response
Host Strain:
rhesus monkey (Macaca mulatta)
Vaccination Protocol:
10^11 vp of Ad5/35-HIV vector was injected i.m. into two rhesus monkeys (2 years old, male) at weeks 0 and 8 (Xin et al., 2005).
Immune Response:
A detectable HIV-specific serum Ab response developed within 2 weeks of the first immunization. At 4 weeks post boosting, titers in excess of 1:50 000 were achieved. Similar results were observed in neutralizing Ab. A increase in the number of HIV-specific IFN-gamma-secreting T cells was also detected in the peripheral blood mononuclear cells (PBMCs). Boosting with Ad5/35-HIV vector further increased this T-cell response (Xin et al., 2005).
Side Effects:
Liver infection with Ad5 vector was 20- to 40-fold stronger than that with Ad5/35 vector. Ad5-Luc vector was two- and four-fold higher, respectively, than that of the monkeys that received the Ad5/35-Luc vector. The Ad5/35 recombinants exhibits minimal hepatotoxicity in non-human primates but is also significantly less susceptible to the pre-existing Ad5 immunity than a comparable Ad5 vector (Xin et al., 2005).
l.
Mouse Response
Vaccination Protocol:
Mice were injected i.m. with Ad5-Luc or Ad5/35-Luc. Luciferase expression was monitored using an in vivo imaging system (IVIS). The expression of HIV gp160 was confirmed by Western blotting. Mice were immunized with Ad5/35-HIV vector, and the HIV-specific CMI was periodically monitored by the intracellular cytokine staining (ICS) assay (Xin et al., 2005).
Immune Response:
The animals immunized with Ad5/35-HIV vector developed a high-titered anti-gp160 antibody (Ab) response. The magnitude of this response was not significantly altered by preimmunization with the DNA-HIV vaccine. DNA-HIV vaccination alone generated a low level of HIV-specific serum Ab. HIV-specific neutralizing Ab was only detectable in the Ad5/35-HIV vaccinated mice and DNA prime/Ad5/35-HIV boosted mice. HIV-specific cellular immune responses persisted through 7 months after final immunization (Xin et al., 2005).
Side Effects:
The hepatotoxicity caused by the Ad5 vector was circumvented by the use of an Ad5/35 vector (Xin et al., 2005).
Challenge Protocol:
Immunized mice were challenged with vPE16 2 weeks after final immunization. Vaccinated mice were challenged with vPE16 7 weeks after final immunization. The strain vPE16 is HIVBH8 gp160-expressing replication-competent vaccinia virus (WR strain, vPE16; HIVBH8 gp160 has 97.32% amino-acid homology with HIVIIIB gp160) (Xin et al., 2005).
Efficacy:
The animals that were vaccinated with the Ad5/35 vector alone or in combination with the DNA-HIV vaccine were completely protected from infection; however, the DNA-HIV vaccination alone had little impact on the susceptibility to infection by vPE16. DNA-HIV vaccination by itself was not protective, but the combination of DNA-HIV priming and Ad5/35-HIV boosting yielded a prolonged and complete protection (Xin et al., 2005).
Vaccination Protocol:
The potency of the vaccine was determined by injecting mice with serial dilutions of gp160 alone or adjuvanted with different formulations. Groups of 10 mice were injected subcutaneously with 1 ml of fourfold dilutions of the test substance. A total of 50 mice were injected in each test (Barrett et al., 1989).
Immune Response:
The highest potency in mice was obtained using a preparation with 0.2% Al(OH)3 and 0.25% deoxycholate (Barrett et al., 1989).
21. HIV recombinant vector vaccine MVA-gag encoding gag
Immune Response:
Immunization of macaques with MVA-gag resulted in relatively weak antigen-specific T-cell responses; the levels did not exceed 150 spot-forming cells (SFC)/106 PBMCs after three doses and were significantly less than those observed in the Ad5-gag vaccinees. Only one of six MVA-gag vaccinees elicited any detectable Gag-specific antibody response (140 mMU/ml at 4 weeks post-dose 3 for monkey V215) (Casimiro et al., 2003).
Efficacy:
The study reports that SIV Gag delivered by DNA, MVA, or adenovirus type 5 vectors is able to inhibit viral replication and disease progression in rhesus macaques following challenge with the SHIV89.6P virus. However, only 50% of the animals that received MVA alone or in combination with DNA were able to effectively control viremia (Casimiro et al., 2003).
22. HIV recombinant vector vaccine MVA.HIVA encoding env and rev
Vaccination Protocol:
Rhesus macaques were divided into four groups and nasally immunized with vaccine containing: 1) 100 µg of gp120 alone, 2) 100 µg of gp120 plus 10 µg of nCT, 3) 100 µg of gp120 plus 25 µg of mCT E112K, or 4) 100 µg of gp120 plus 100 µg of mCT E112K. Macaques were anesthetized with ketamine and placed in dorsal recumbancy with head tilted back so that the nares were pointed upward. Vaccine solution (0.5 ml) was instilled dropwise into each nostril without inserting the syringe into the nasal cavity. Macaques were kept in that position for 10 min and then placed in lateral recumbancy until they recovered from anesthesia, as described previously (16). Nasal immunization was conducted on days 0, 7, 14, 28, 42, and 56 (Yoshino et al., 2004).
Immune Response:
Macaques given nasal gp120 with either mCT E112K or nCT showed elevated gp120-specific IgG and IgA Ab responses with virus-neutralizing activity in both their plasma and mucosal external secretions, as well as higher numbers of gp120-specific IgA Ab-forming cells in their mucosal and peripheral lymphoid tissues and of IL-4-producing Th2-type CD4-positive (CD4(+)) T cells than did controls. Even though significant mucosal adjuvanticity was seen with both mCT E112K and nCT, neuronal damage was observed only in the nCT-treated, but not in the control or mCT E112K-treated groups (Yoshino et al., 2004).
Host Gene Response of
IgA Fc fragment
Gene Response:
Macaques given nasal gp120 with either mCT E112K or nCT showed significantly elevated IgA Ab responses with virus-neutralizing activity in both their plasma and mucosal external secretions, as well as higher numbers of gp120-specific IgA Ab-forming cells in their mucosal and peripheral lymphoid tissues compared to controls, who were vaccinated without the adjuvant (Yoshino et al., 2004).
Gene Response:
Macaques given nasal gp120 with either mCT E112K or nCT showed significantly elevated gp120-specific IgG responses with virus-neutralizing activity in both their plasma and mucosal external secretions compared to controls, who were vaccinated without the adjuvant (Yoshino et al., 2004).
Gene Response:
Vaccinated animals that received the adjuvant had a greater increase in IL-4-producing Th2-type CD4-positive (CD4(+)) T cells in mesenteric lymph nodes (MLNs) than did controls vaccinated without the adjuvant, which did not produce IL-4 (Yoshino et al., 2004),
The DNA vaccine constructs were developed based on the gene sequences of the gp140 envelope, p55 Gag, Nef, and Tat proteins from the HIV-2UC2 isolate (GenBank accession number U38293) (Locher et al., 2004).
Vaccination Protocol:
Each baboon was immunized four times with the experimental or control DNA constructs formulated with Vaxfectin. The time intervals were chosen to permit maturation of the immune response (Locher et al., 2004).
Challenge Protocol:
One month after the fourth immunization, the baboons were challenged twice (6 h apart) by the intravaginal route using 50 baboon infectious dosages (30,000 tissue culture infectious dosages) of HIV-2UC2/9429 for a total of 100 baboon infectious dosages. This virus isolate is a more virulent variant of the HIV-2UC2 isolate and was recovered from a baboon with an AIDS-like disease (Locher et al., 2004).
Efficacy:
At 2 weeks post-HIV-2 challenge, viral loads ranged from 100 (below the threshold of detection) to 81,000 viral RNA copies in the plasma, with the lowest average viral loads in the group of baboons that received the HIV-2 DNA without the genetic adjuvants (a mean of 852 copies/ml), followed by the group of baboons the received the HIV-2 DNA and the genetic adjuvants (a mean of 2499 copies/ml) (Locher et al., 2004).
Vaccination Protocol:
Mice received 0.5 ml of the specified vaccine preparations subcutaneously under the abdominal skin on day 0 (Stieneker et al., 1991).
Immune Response:
Mice immunized with PMMA adjuvant had higher antibody counts than mice immunized with an alum adjuvant (Stieneker et al., 1991).
A non-pathogenic protozoan parasitic vector, Leishmania tarentolae, which shares common target cells with HIV-1, was used to express full-length HIV-1 Gag protein (Breton et al., 2007).
h. Immunization Route
Intramuscular injection (i.m.)
i.
Mouse Response
Host Strain:
Balb/c
Vaccine Immune Response Type:
VO_0003057
Immune Response:
Immunization of BALB/c mice with recombinant L. tarentolae led to the expansion of HIV-1 Gag-specific T cells and stimulated CD8(+) T cells to produce gamma interferon in response to specific viral Gag epitopes. A booster immunization with recombinant L. tarentolae elicited effector memory HIV-1 Gag-specific CD4(+) T lymphocytes and increased antibody titres against HIV-1 Gag (Breton et al., 2007).
Efficacy:
Immunization of human tonsillar tissue cultured ex vivo with Gag-expressing L. tarentolae vaccine vector elicited a 75% decrease in virus replication following exposure of the immunized tonsils to HIV-1 infection (Breton et al., 2007).
An rMVA vaccine that expresses HIV Gag, Pol, and Env proteins was constructed (Liu et al., 2006).
f.
Mouse Response
Host Strain:
BALB/c
Vaccination Protocol:
Female BALB/c mice were used for immunizations. All were conducted on anesthetized mice. Immunizations with MVAs were accomplished by inoculating the desired amount of MVA into a single quadriceps muscle. Immunizations with DNA were accomplished by injecting the desired amount of DNA into the quadriceps, half in each leg (Liu et al., 2006).
Persistence:
Temporal CD8 responses were conducted in BALB/c mice using MVA. Responses were steady for >10 weeks (>6 weeks post-MVA) (Liu et al., 2006).
Immune Response:
CD8 T cells were boosted more effectively than CD4 T cells with the ratio of elicited CD8 to CD4 cells for the immunodominant CD8 epitope in Gag increasing with boosts. The most effective boost for CD8 T cells resulted in the greatest skewing of the CD8 to CD4 T cell ratio. This could represent better access of CD8 than CD4 T cells to APCs (Liu et al., 2006). The dose–response studies showed good increases for antigen expression with increasing MVA dose. A 1000-fold increase in the dose of MVA resulted in a 300-fold increase in the frequency of antigen-expressing cells. In contrast, dose–response studies for in vivo immunogenicity showed <10-fold increases in elicited T cells and Ab for 100–1000-fold increases in the dose of inoculated MVA. Shallow dose–response curves for immunogenicity were observed post priming as well as post boosting of an MVA or a DNA prime (Liu et al., 2006).
Side Effects:
No adverse effects were encountered (Liu et al., 2006).
Attenuated poxvirus-HIV-1 recombinants followed by protein boosting (Patterson et al., 2000).
f. Immunization Route
Intramuscular injection (i.m.)
g.
Macaque Response
Vaccination Protocol:
Twenty-four macaques were primed with NYVAC (a genetically attenuated Copenhagen vaccinia strain) recombinants with HIV-1 and HIV-2 env and gag-pol or NYVAC vector alone and boosted with homologous, oligomeric gp160 proteins or adjuvant only (Patterson et al., 2000).
Vaccine Immune Response Type:
VO_0000287
Challenge Protocol:
One half of each immunization and control group were intravenously challenged with SHIV(HXB2) the other half was challenged with HIV-2SBL6669 (Patterson et al., 2000).
Efficacy:
Virological assessments verified that both NYVAC-HIV-1 and NYVAC-HIV-2 immunization significantly reduced viral burdens and partially protected against HIV-2 challenge, although cross-protection was not at the level that had been previously reported. Humoral antibody and/or CTL and CD8AA were associated with protection against homologous HIV-2 challenge, while cellular immune responses seemed more important for cross-protection. No significant protection was observed in the SHIV-challenged macaques (Patterson et al., 2000).
Attenuated poxvirus-HIV-1 recombinants followed by protein boosting (Patterson et al., 2000).
f. Immunization Route
Intramuscular injection (i.m.)
g.
Macaque Response
Vaccination Protocol:
Twenty-four macaques were primed with NYVAC (a genetically attenuated Copenhagen vaccinia strain) recombinants with HIV-1 and HIV-2 env and gag-pol or NYVAC vector alone and boosted with homologous, oligomeric gp160 proteins or adjuvant only (Patterson et al., 2000).
Vaccine Immune Response Type:
VO_0003057
Challenge Protocol:
One half of each immunization and control group were intravenously challenged with SHIV(HXB2) the other half was challenged with HIV-2SBL6669 (Patterson et al., 2000).
Efficacy:
Virological assessments verified that both NYVAC-HIV-1 and NYVAC-HIV-2 immunization significantly reduced viral burdens and partially protected against HIV-2 challenge, although cross-protection was not at the level that had been previously reported. Humoral antibody and/or CTL and CD8AA were associated with protection against homologous HIV-2 challenge, while cellular immune responses seemed more important for cross-protection. No significant protection was observed in the SHIV-challenged macaques, although NYVAC-HIV-1 immunization resulted in significantly lower viral burdens compared with controls (Patterson et al., 2000).
The gag, pol, and env genes of SIV(K6W) were expressed in the NYVAC vector (Benson et al., 1998).
f. Immunization Route
Intramuscular injection (i.m.)
g.
Macaque Response
Vaccination Protocol:
The macaques were immunized with NYVAC-SIV and NYVAC-IL-12 or NYVAC-SIV alone (Benson et al., 1998).
Vaccine Immune Response Type:
VO_0003057
Challenge Protocol:
At the end of the immunization regimen, half of the animals were challenged with SIV(mac251) by the intravenous route and the other half were exposed to SIV(mac251) intrarectally (Benson et al., 1998).
Efficacy:
Significantly, five of the eleven vaccinees exposed mucosally to SIV(mac251) showed a transient peak of viremia 1 week after viral challenge and subsequently appeared to clear viral infection. In contrast, all 12 animals inoculated intravenously became infected, but 5 to 6 months after viral challenge, 4 animals were able to control viral expression and appeared to progress to disease more slowly than control animals. Protection did not appear to be associated with any of the measured immunological parameters. Further modulation of immune responses by coadministration of NYVAC-cytokine recombinants did not appear to influence the outcome of viral challenge (Benson et al., 1998).
31. rBCG-SIVgag and rDIsSIVgag Prime-boost SHIV vaccine
Description:
The SHIV gag gene was inserted to a recombinant BCG vector and a replication-deficient vaccinia virus strain (DIs) vaccine vector (Ami et al., 2005).
Mycobacterium bovis bacillus Calmette-Guérin (BCG)-Tokyo and a replication-deficient vaccinia virus strain (DIs) (Ami et al., 2005)
h. Immunization Route
Intramuscular injection (i.m.)
i.
Macaque Response
Host Strain:
cynomologus macaque
Vaccination Protocol:
cynomologus macaques were primed with rBCG-SIVgag, and then boosted with rDIsSIVgag (Ami et al., 2005).
Vaccine Immune Response Type:
VO_0003057
Challenge Protocol:
Ten weeks after the second booster immunization, the macaques were challenged by intrarectal inoculation with 2 × 10^3 TCID50 or 50 50% monkey infectious doses (MID50) of SHIV KS661c, an SHIV-89.6 variant clone (Ami et al., 2005).
Efficacy:
For the prime-boost vaccination group, plasma viremia levels remained undetectable and CD4+ T-cell counts stayed above 500 cells/μl for the entire year of testing (Ami et al., 2005).
32. Recombinant HIV gp120 with adjuvant NanoEmulsion
Description:
An oil-in-water nanoemulsion (NE) was used as the mucosal adjuvant for this vaccine (Bielinska et al., 2008).
e. Preparation
The gp120 and NE vaccines were prepared by mixing NE with gp120 protein solution, using pyrogen-free saline as a diluent (Bielinska et al., 2008).
f.
Mouse Response
Host Strain:
BALB/c
Vaccination Protocol:
BALB/c mice were immunized with two or three intranasal administrations of gp120/NE formulation at 3 weeks apart. The immunizations of 10 microliters were performed by slowly applying gp120/NE mixes to the nares.Control mice were immunized with gp120 in saline, with NE alone or saline. Intramuscular immunization was performed with two doses, 3 weeks apart, of 20 micrograms of gp120 injected in 50 microliters of either saline or 1% NE (Bielinska et al., 2008).
Immune Response:
Immunized mice demonstrated robust serum anti-gp120 IgG, as well as bronchial, vaginal, and serum anti-gp120 IgA . The analysis of gp120-specific CTL proliferation, INF- induction, and prevalence of anti-gp120 IgG2 subclass antibodies indicated that nasal vaccination in NE also induced systemic, Th1-polarized cellular immune responses (Bielinska et al., 2008).
g.
Guinea pig Response
Host Strain:
Hartley
Vaccination Protocol:
Hartley guinea pigs were immunized intranasally with two administrations (50 microliters per nare) of gp120/NE mix at 3 weeks apart (Bielinska et al., 2008).
Immune Response:
Immunization produced significant levels of serum anti-gp120 IgG antibodies in all animals, as was seen with the mice. The analysis of gp120-specific CTL proliferation, INF gamma induction, and prevalence of anti-gp120 IgG2 subclass antibodies indicated that nasal vaccination in NE also induced systemic, Th1-polarized cellular immune responses (Bielinska et al., 2008).
Vaccination Protocol:
Groups of five mice were immunized subcutaneously three times at 14-day intervals. The immunoliposomes were suspended in PBS and each mouse received 0.1 ml per injection. Each injection contained 380 μg of PC, 193 μg of CH, 10 pg of PE-B, 2.0 μg of MAb, and 10 μg of rgpl20 (Ozpolat et al., 1998).
Immune Response:
Mice vaccinated with immunoliposomes were found to have a strong delayed-type hypersensitivity (DTH) response to the weakly immunogenic gp120 that was dependent on the presence of the MAbs. However, this vaccination protocol did not induce humoral immunity (Ozpolat et al., 1998).
Co-expressed the CD40L with our DNA/SIV vaccine such that the CD40L is anchored on the membrane of SIV virus-like particle (VLP) (Kwa et al., 2014).
f. Immunization Route
Intramuscular injection (i.m.)
g.
Macaque Response
Vaccination Protocol:
The macaques were divided into 3 groups and group 1 was vaccinated with a DNA/MVA SIV vaccine, group 2 was vaccinated with the DNA/MVA SIV vaccine with CD40L in the DNA, and the third group was an unvaccinated control (Kwa et al., 2014).
Vaccine Immune Response Type:
VO_0003057
Challenge Protocol:
The macaques were intrarectally challenged with SIVE660 at a dose of 5000 TCID to test the level of protection of the vaccines (Kwa et al., 2014).
Efficacy:
CD40L adjuvant enhanced the functional quality of anti-Env antibody response and breadth of anti-SIV CD8 and CD4 T cell responses, significantly delayed the acquisition of heterologous mucosal SIV infection and improved viral control. Notably, the CD40L adjuvant enhanced the control of viral replication in the gut at the site of challenge that was associated with lower mucosal CD8 immune activation, one of the strong predictors of disease progression (Kwa et al., 2014).
Description:
A recombinant modified replicating vaccinia virus Tiantan strain (MVTTSIVgpe) and a recombinant, nonreplicating adenovirus type 5 strain (Ad5SIVgpe) expressing the SIVmac239 Gag, Pol, and Env structural proteins were generated (Sun et al., 2013).
Description:
A recombinant modified replicating vaccinia virus Tiantan strain (MVTTSIVgpe) and a recombinant, nonreplicating adenovirus type 5 strain (Ad5SIVgpe) expressing the SIVmac239 Gag, Pol, and Env structural proteins were generated (Sun et al., 2013).
Description:
A recombinant modified replicating vaccinia virus Tiantan strain (MVTTSIVgpe) and a recombinant, nonreplicating adenovirus type 5 strain (Ad5SIVgpe) expressing the SIVmac239 Gag, Pol, and Env structural proteins were generated (Sun et al., 2013).
(Sun et al., 2013) a modified replicating vaccinia virus Tiantan strain (MVTT(SIVgpe))
i. Immunization Route
Intramuscular injection (i.m.)
j.
Monkey Response
Vaccination Protocol:
Eight monkeys were divided into two groups: (i) four monkeys received the MVTTioin+Adim testing regimen as in study I, and (ii) four monkeys received an empty MVTT control vector (10^9 PFU) through intraoral (0.5 ml) and intranasal (0.5 ml) routes and an empty Ad5 control vector (10^11 vp in 1 ml of PBS) through intramuscular injection (Sun et al., 2013).
Vaccine Immune Response Type:
VO_0003057
Challenge Protocol:
At either week 30 after the initial vaccination or week 24 after the final vaccination, each animal was challenged intrarectally with 5 × 10^5 50% tissue culture infective doses (TCID50) of Chinese rhesus monkey-adapted and neutralization-resistant SIVmac239. In all cases, the challenge virus stock was administered in 1 ml of PBS (Sun et al., 2013).
Efficacy:
The reductions in peak and set-point viral loads were significant in most animals, with one other animal being protected fully from high-dose intrarectal inoculation of SIV(mac239). Furthermore, the animals vaccinated with this regimen were healthy, while ~75% of control animals developed simian AIDS. The protective effects correlated with the vaccine-elicited SIV-specific CD8(+) T cell responses against Gag and Pol (Sun et al., 2013).
Efficacy:
The immune response elicited by the multivalent DNA prime/protein boost vaccine was able to protect macaques from rectal challenge with SHIV-Ba-L isolate. This polyvalent vaccine formulation was able to confer protection in four out of six animals against SHIV-Ba-L and significantly reduced plasma viremia in the two remaining animals (Pal et al., 2006).
Efficacy:
one of six immunized animals was completely protected whereas all six naïve animals were infected. These results demonstrate that a vaccine based on priming with a polyvalent DNA vaccine from multiple HIV-1 subtypes followed by boosting with homologous Env proteins elicits anti-HIV-1 immune responses capable of controlling rectal transmission of SHIV(Ba-L) (Pal et al., 2005).
Vaccination Protocol:
Mice were immunized with 10 μg of V3-VLPs without adjuvant in the muscle of the left leg. At the same time, the mice were immunized in the right leg with the other immunogens (Harris et al., 1996).
Immune Response:
Mice immunized with V3 VLPs adjuvanted with gamma inulin showed good CTL responses (Harris et al., 1996).
HIV-1 p24 (clade B consensus), was inserted near the 5' end of YF17D, in frame and upstream of the polyprotein (YF-5'/p24) (Franco et al., 2010).
f. Immunization Route
Intramuscular injection (i.m.)
g.
Mouse Response
Vaccination Protocol:
Mice were immunized two times subcutaneously at the base of the tail with PBS or 10^6 pfu of YF17D, YF-E/p24/NS1, or YF-5′/p24 in 100 μl on days 0 and 14 (Franco et al., 2010).
Vaccine Immune Response Type:
VO_0003057
Challenge Protocol:
Three weeks after the second immunization, half of the mice in each group were challenged intranasally with 5 × 10^5 pfu of Gag-expressing recombinant vaccinia virus (Vac-gag). The remaining animals in the groups were challenged intranasally with the same dose of wild-type vaccinia virus (Vac-wt) (Franco et al., 2010).
Efficacy:
The protective efficacy of the YF17D recombinants, particularly YF-E/p24/NS1, in mice challenged with a vaccinia expressing HIV-1 Gag was demonstrated (Franco et al., 2010).
V. References
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5. Bagley et al., 2003: Bagley KC, Shata MT, Onyabe DY, DeVico AL, Fouts TR, Lewis GK, Hone DM. Immunogenicity of DNA vaccines that direct the coincident expression of the 120 kDa glycoprotein of human immunodeficiency virus and the catalytic domain of cholera toxin. Vaccine. 2003; 21(23); 3335-3341. [PubMed: 12804865].
6. Barouch et al., 2012: Barouch DH, Klasse PJ, Dufour J, Veazey RS, Moore JP. Macaque studies of vaccine and microbicide combinations for preventing HIV-1 sexual transmission. Proceedings of the National Academy of Sciences of the United States of America. 2012; 109(22); 8694-8698. [PubMed: 22586094].
7. Barrett et al., 1989: Barrett N, Mitterer A, Mundt W, Eibl J, Eibl M, Gallo RC, Moss B, Dorner F. Large-scale production and purification of a vaccinia recombinant-derived HIV-1 gp160 and analysis of its immunogenicity. AIDS research and human retroviruses. 1989; 5(2); 159-171. [PubMed: 2713166].
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