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REVIEW Gene-Mutated HIV-1/SIV Chimeric Viruses As AIDS Live Attenuated Vaccines for Potential Human

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REVIEW Gene-Mutated HIV-1/SIV Chimeric Viruses As AIDS Live Attenuated Vaccines for Potential Human Leukemia (1999) 13, Suppl. 1, S42–S47 1999 Stockton Press All rights reserved 0887-6924/99 $12.00 http://www.stockton-press.co.uk/leu REVIEW Gene-mutated HIV-1/SIV chimeric viruses as AIDS live attenuated vaccines for potential human use M Hayami1, T Igarashi1, T Kuwata1,MUi1, T Haga1, Y Ami2, K Shinohara2 and M Honda2 1Institute for Virus Research, Kyoto University, Kyoto; and 2AIDS Center, Division of Experimental Animal Research, National Institute of Infectious Diseases, Japan To develop an AIDS vaccine for human use as well as a suitable T cells against HIV-1 Env without any clinical symptoms; (2) animal model for AIDS research, we constructed a series of SIV Gag crossreacts with HIV-1 Gag, which can be another HIV-1/SIVmac chimeric viruses (SHIVs). We successfully gener- ated a SHIV (designated as NM-3rN) having the HIV-1 env gene, neutralizing epitope besides Env; (3) SIV is genetically close which enabled the evaluation of the efficacy of HIV-1 Env-tar- to HIV-2 which is less pathogenic than HIV-1, and accidental geted vaccines in macaque monkeys instead of chimpanzees. SIV infection in humans resulted in silent infection, suggesting Two NM-3rN derivatives (NM-3 and NM-3n) induced long-term that SIV is not pathogenic in humans; (4) these SHIVs can anti-virus immunities without manifesting the disease. The infect human lymphocytes, suggesting that the infection monkeys vaccinated with NM-3 or NM-3n became resistant to immunity by SHIVs may be induced in humans and protect a challenge inoculation with NM-3rN. Serum from a monkey vaccinated with NM-3 neutralized not only the parental HIV-1 humans from the HIV-1 infection. For the purpose of making (NL432), but also an antigenically different HIV-1 (MN). In vivo a vaccine, therefore, some genes essential for pathogenicity experiments confirmed the heterologous protection against an were mutated to attenuate these chimeric viruses. We SHIV having the HIV-1 (MN) env. In addition to specific immun- observed that the monkeys infected with these chimeric ity including neutralizing antibodies and cytotoxic T lympho- viruses were protected against a challenge infection. In this cyte activity, nonspecific immunity such as natural killer report, we present recent progress in our studies of SHIVs, and activity is associated with this protection. These data suggest that the live vaccine has the ability to protect individuals discuss their applications to the development of an AIDS against various types of HIVs. These SHIVs should contribute vaccine. to the development of future anti-HIV-1 live vaccines in humans. Keywords: HIV-1; SIV; chimeric virus; AIDS; live vaccine Strategy for constructing chimeric viruses First, each gene of HIV-1 and SIV was investigated by con- structing various mutant viruses and determining their viral Introduction replication capacity and their exchangeability between the two viruses.6,7 As a result, the strategy for construction of The lack of a suitable animal model for evaluating vaccine chimeric viruses between HIV-1 and SIV was determined as candidates of human immunodeficiency virus type 1 (HIV-1) follows: (1) vpr and nef can be sacrificed; (2) tat and TAR (in is a difficult problem in developing the vaccine, because few LTR) can be from different parental viruses, but rev and RRE animal species are susceptible to HIV-1 infection. Pig-tailed (in env), which interact, should have the same origin; (3) gag macaques1 and rabbits2 were reported to be infected with and env can be from different origins, but gag and pol should HIV-1, but they could not be used practically. At present, be from the same origin. chimpanzees are the only animals used for evaluation of HIV- Based on this strategy, we constructed chimeric viruses 1 vaccines, although some human trials have been conduc- between HIV-1 and SIVmac. SIVmac is known to induce an ted.3 Chimpanzees, however, rarely show any AIDS-like dis- AIDS-like disease in macaque monkeys. At first, chimeric ease, and their use is extremely limited for experiments viruses were constructed by replacing the 5′-half containing because they are an endangered species. We therefore tried gag and pol, and the 3′-half containing env, tat and rev to generate an HIV-1 that is infectious to, and able to induce, between HIV-1 and SIVmac, and their replicative abilities an AIDS-like disease in macaque monkeys, by constructing were investigated in human or monkey peripheral blood HIV-1 chimeric viruses based on SIVmac (SHIVs). As a result, mononuclear cells (PBMC). As a result, the chimeric viruses we succeeded in constructing chimeric viruses, which include that had the env of HIV-1 and the gag and pol of SIVmac the env gene of HIV-1 and which could infect macaque mon- could replicate in monkey PBMC, but the other viruses, which keys, a species that is available for experimental use.4,5 These had the env of SIVmac and the gag and pol of HIV-1, could chimeric viruses express the HIV-1 Env protein, and enable not4 (Figure 1). This result shows that the gag and pol genes us to use macaque monkeys instead of chimpanzees for evalu- of SIVmac determine the tropism to monkey PBMC and that ation of vaccine candidates. This is done by vaccinating the the env gene has no relation to this cell tropism. This fact was monkeys with a vaccine candidate and then challenging them unexpected because Env, whose function is to bind to cells, with the chimeric viruses. was generally supposed to determine the cell tropism. In addition, SHIVs may be used as anti-HIV-1 live attenu- ated vaccines for humans for the following reasons: (1) SHIV- infected monkeys produced neutralizing antibodies and killer Experimental infection of monkeys with HIV-1/SIVmac chimeric viruses 5,8 Correspondence: M Hayami, Institute for Virus Research, Kyoto Uni- In the series of chimeric viruses constructed, NM-3 and versity, Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606, Japan NM-3n,9 which could replicate in monkey PBMC, were HIV-1/SIVmac chimeric viruses M Hayami et al S43 Figure 1 Replication and pathogenicity of HIV-1, SIVmac and HIV-1/SIVmac chimeric viruses in vitro and in vivo. inoculated to cynomolgus monkeys intravenously (i.v.). Their (CTL) were detected at 3 years post-inoculation. In the other abilities to infect monkeys were examined by virus recovery, monkey, proviral DNA was detected in PBMC only in the the polymerase chain reaction (PCR) and antibody response early period after inoculation, and anti-Env antibodies were (Table 1). NM-3 has LTR, gag, pol, vif and vpx of SIVmac and detected for a while. Thus, this monkey was considered to be env, tat, rev and vpu of HIV-1. NM-3n, which has a similar infected transiently.5 These results showed that NM-3 infec- structure but contains the intact nef gene was constructed tion induced both humoral and cellular anti-virus immunities based on the report by Kestler et al10 who demonstrated that in monkeys, although its infectivity was not so efficient. the nef gene was required for the induction of AIDS-like Two monkeys (a 3-year-old male and a 3-year-old female) 3.5 disease in monkeys infected with SIVmac. were inoculated with 10 TCID50 of NM-3n. Viruses were Of two monkeys (8- and 11-year-old males) inoculated with occasionally re-isolated from one of these monkeys (male), 4 10 TCID50 of NM-3, one developed a persistent viremic state, and proviruses were also detected in its PBMC by PCR in the and anti-Env and anti-Gag antibodies were raised as expected early period after inoculation. Antibodies which neutralized from the structure of this chimeric virus. These antibodies had NM-3n and the parental HIV-1 were detected persistently in virus neutralizing activity, which neutralized NM-3 and the monkeys infected with NM-3n. HIV-1 Env-specific killer T parental HIV-1. HIV-1 Env-specific cytotoxic T lymphocytes cells were observed at 2 years post-inoculation in the monkey Table 1 Summary of experimental infection of NM-3 and NM-3n Animal Experimental infection with NM-3 (weeks) 0 2 6 10 21 28 40 94 100 116 129 138 149 virus recovery MF1 −++−−−−++++++ MF2 −−−−−−−−−−−−− antibodies (Western) anti-HIV-1 Env MF1 −−−++++++++++ MF2 −−++++−ND ND ND ND ND ND anti-SIVmac Gag MF1 −−+++++++++++ MF2 −−−−−−−ND ND ND ND ND ND Experimental infection with NM-3n (weeks) 0 2 6 10 12 14 20 30 37 42 52 63 73 88 virus recovery MF3080 −−−−−−−−−−−−−− MF3096 −−−−−−−−+−−+−− PCR (env) MF3080 −++−−−−−ND −−−−ND MF3096 −++−+++−ND −−−−ND antibodies (Western) anti-HIV-1 Env MF3080 − ND ND + ND +++++++++ MF3096 − ND ND − ND +++++++++ anti-SIVmac Gag MF3080 − ND ND + ND +++++++++ MF3096 − ND ND − ND −−−++++++ ND, not done. HIV-1/SIVmac chimeric viruses M Hayami et al S44 from which the virus was re-isolated at 63 weeks post-inocu- keys died of an AIDS-like syndrome, from which we are now lation (w.p.i.).9 These results showed that NM-3n, as well as trying to clone the pathogenic chimeric virus. NM-3, infected monkeys and induced both humoral and cellular anti-virus immunities. Viruses re-isolated from the monkeys infected with NM-3 Development of vaccines using chimeric viruses having or NM-3n basically maintained their chimeric structure, HIV-1 Env which had Env of HIV-1 and Gag of SIVmac. Anti-HIV-1 vaccines including inactivated viral particles, recombinant viral proteins and synthetic peptides, are cur- Construction of a chimeric virus containing vpr and rently on trial in various laboratories. They are so-called ‘inac- nef, which produces a more efficient infection tivated-vaccines’ which cannot replicate, and are not thought to be very efficient.
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