Jpn. J. Infect. Dis., 55, 2002

Laboratory and Epidemiology Communications

lnduction of HIV‑ 1 ‑Specific Neutralizing Antibodies in Mice Vaccinatedwith a Recombinant Sendai Virus Vector
Munehide Kanol, Tetsuro Matanol,2串, Atsushi Kato3,4, Tatsuo Shioda4,5and Yoshiyuki Nagail,4,6
1AIDS Research Center and3Department of Viral Diseases and Vaccine Control, National Institute of Infectious Diseases,

Gakuen 4‑7), Musashi‑Murayama, 7Tob)0 208‑0011,
2Department of Microbiology, GradLiate School of Medicine, Universio7 0f Tolg)0,
Hongo 7‑3‑1, Bunlg,0‑ku, Too)0 113‑0033,
4Department of mral Infection, InstittLte OfMedical Science, Universio7 0f 7Tolg)0,
Shirokanedai 4‑6‑1, Minato‑ku, ToIg)0 1 08‑8539,
5Department of Viral Infections, Research lnstitute for Microbial Diseases, Osaka Universio?,
Yamadaoka 1 7‑13‑1, Suita, Osaka 565‑0871 and6Toyama Institute ofHealth,

Nakataikou‑yama 1 7‑1, Kosugi, Imizu, Toyama 939‑0363
Communicated by Naoki Yamamoto(Accepted June 7, 2002)

have been studying a re90mbinant Sendai virus (SeV) Vector

recovered as described previously (5). The SeV/gp160
expressesfull length HIV‑1NL4̲3 Env, gp1 60. The SeV/gp140
expresses the extracellular portion of HIV‑ 1 NL4̲3 Env, gp140,

system as an AIDS vacclne Candidate (1‑4). We constmcted arecombinant SeV vector expresslng Simian immunodeficiency

consistlng Ofthe whole gp120 and the gp41 ectodomains.
Balb/c mice were intranasally Immunized with 107 cellvirus (SIV) Gag protein and showed its potential for efFiciently

infectious units of SeV/gp1 60 or SeV/gp140 twice at weeks

VinlS‑SpeC捕c CD8+ T cells and neutralizing antibodies

are important effectors against Virus infections in the host. We

inducing antigenIspecific CD8+ T cells in macaques (3,4)･

0 and 2. The immunized mice were sacrificed 4 weeks a洗er

In this study, uslng a mouse model, We examined if thethe first immunization, and the sera were prepared from therecombinant SeV vector‑based vaccine system can induce

whole bloods, inactivated at 56oC for 30 min, and subjectedvirus‑Specific neutralizing antibodies as well.

to neutralization assay.

Two kinds of SeV vectors expressing HIVl envelope

Tbe HIV‑lNL4̲3 Stock solutions diluted serially fburfbld in

glycoproteins (Env), SeV/gp160 and SeV/gp140, werequadricate were incubated with an equal volume of theinactivated control sera or the sera prepared from the SeV/
*Corresponding author: Tel: +8 1 ‑3‑5841‑3407, Fax: +8 1‑3‑584 I

gp1 60‑ or SeV/gp140‑immunized mice. After incubation at3374, E‑mail: matano@m.u‑tokyo.ac,jp

room temperature for 1 h, the mixtures were used to infect59

Jpn. J. Infect. Dis., 55, 2002
Table I. HIV‑1‑specific neutralizing activities in immunizedmice
Infectious titer in the presence%illllibiti on

ofthe indicated semm
TC ID5 0/ml 1 ogarithm
Control Semm
lst

5.16E+05

5.71

2nd2.06E十06

6,31

Geometric mean (M) 1.03E+06

6.01

SD0.43

M‑2SD

5.15

Serum from SeV/gp140‑immunizedmice#1

7.05E+04#2

1.13E+06#3

1.29E+05#4

7.76E+04

4 ′0 5 4 8 0 1 8 5 5 1 ∩7** *0ノ nXU (uノ3 0 7 2 2055

Serum from SeV/gp1 60‑immunizedmice′LU ′hU ′hU ′LU 5 ′0

#55.20E十06#6

1.13E+06#7

3.28E+06#82.06E+06#95.16E十05#10

2.38E+06

7 0 5 3 7 3 2 5 2 1 1 nXU

0 0 0 0 07 0 0 0 0 0 人uノ 0

4The %inhibition was calculated as (1‑[HIV一日iter in the presence oftlle Senlm]/【HIVltiter in the prPS.e.nee Ofthe control serum]) × 100 (%)･ The asterisk indicates signiflCanCe Of

neutralizing actlVltleS (<M‑2SD).

MT4 cells cultured in 96‑well plates at a concentration ofgenesis. Reverse genetics transfbms understanding. Rev.

Med. Viro1., 9, 83‑99.
2. Kano, M., Matano, T., Nakamura, H., Takeda, A., Kato,A., Ariyoshi, K., Mori, K., Sata, T. and Nagai, Y.

5 × 1 04 cells per well. ARer 14 days ofculture, the supematants

were harvested and subjected to an enzyme‑linked immuno‑sorbent assay (ELISA) in order to detect HIV‑1 Gag p24
antigen (Retro‑Tek, Cellular Products, Inc., Buffalo, N.Y.),and the infectious virus titer was calculated as described(2000): Elicitation of protective iTmunityagainst simian
immunodeficiencyvirus infection by a recombinantSendai virus expresslng the Gag protein. AIDS, 14, 1281‑

befわre (6). The obtained vinュs liters in each condition andtheir geometric meansare shownin Table 1. Three offour sera

1282.

3. Matano, T., Kano, M., Nakamura, H., Takeda, H. andderived from mice immunized with SeV/gp140 significantly
reduced HIV‑ 1 infectivity. However, none of the sera obtained

Nagai, Y (2001 ): Rapid叩pearanCe OfsecondaIγ immune

fTrom six SeV/gp 1 60‑immunizedmice did so. Antibody levels

responses and protection舟om acute CD4 depletion after

measured by an ELISA using HIV‑1NL4̲3 Env as an antlgen
were similar for SeV/gp140‑immunized and SeV/gp1 60‑in macaques vaccinated with a DNA‑pnme/Sendai virala highly pathogenic immunodeficiency virus challenge

immunized groups (data not shown). Therefore, Env antigen

vector‑boost reglmen. ∫. Viro1., 75, 1 1891‑1 1896.

expressed as gp140 appeared more effective in inducing

4. Kano, M., Matano, T., Kato, A., Nakamura, H., Takeda,

neutralizing antibodies than the membrane‑bound gp 1 60 Env.

A., Suzaki, Y., Ami, Y., Terao, K. and Nagai, Y. (2002):

However, the obtained level of the neutralizing capacity Of

Primary replication of a recombinant Sendai viral vector

in macaques. J･ Gent Virol･, 83, 1377‑1386･
5. Yu, D., Shioda, T., Kato,A., Hasan, M. K., Sakai,Y andthe antibodies was low even with SeV/gp140.

Nagai, Y (1 997): Sendai vinュs‑based expression ofHIV

The authors are grate氏ll to Katsuhiro Kiyotani fわr his

technical advice on animal experiments. This work was

1 gp120: reinfわrcement by the V(‑) verslOn. Genes Cells,

supported by Grants舟om the Ministry ofHealth, Labour and2, 457‑466.

Welfare, by Grants from the Human Sciences Foundation,and by a Grant from the Ministry of Education, Culture,

6. Shibata, R., Maldarelli, F., Siemon, C., Matano, T., Parta,M., Miller, G., Fredrickson, T. and Martin, M. A. (1997):

Infection and pathogenicity of chimeric simian‑humanSports, Science and Technology of Japan.

immunodeflCiency viruses in macaques: determinants ofhigh virus loads and CD4 cell killing. J. Infect. Dis., 176,

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