TY - JOUR
T1 - Construction and in vitro characterization of a molecularly cloned human immunodeficiency virus type 1 library
AU - Mukai, Tetsu
AU - Kurosu, Takeshi
AU - Kinomoto, Masanobu
AU - Komoto, Satoshi
AU - Shiraga, Miki
AU - Auwanit, Wattana
AU - Ikuta, Kazuyoshi
N1 - Funding Information:
We thank Madiha S. Ibrahim for critical reading of the manuscript. This work was supported in part by a Grant-in-Aid for Scientific Research on Priority Areas from the Ministry of Education, Culture, Sports, Science and Technology a Grant-in-Aid for AIDS Research from the Ministry of Health, Labor and Welfare of Japan, and the Japanese Foundation for AIDS Prevention and the annual budget of the Department of Medical Science, Thailand.
PY - 2002/1/15
Y1 - 2002/1/15
N2 - Development of a safe and effective vaccine against human immunodeficiency virus (HIV) is urgent, but many concerns regarding the safety and efficacy of the currently developing vaccines remain. A major hindrance in HIV vaccine development is the genetic diversity, a hallmark of HIV biology, and a poor understanding of how HIV vaccine prevents the emergence of escape variants during infection and progression of AIDS. Here, we developed a method to construct a molecularly cloned viral library. This technique employs a long-range polymerase chain reaction (PCR) to amplify a virtually full-length HIV type 1 (HIV-1) provirus genome from peripheral blood mononuclear cells (PBMCs) infected with CRF01_AE (subtype E) Thai primary isolate. Among randomly selected 93 clones, 41 with a full-length sequence were able to replicate in PBMCs, 5 of which induced strong cytopathic effects. Replication kinetics also showed that the parental virus was intermediate among the clones. Thus, the molecular library prepared by this method showed the quasi-species in infected cells and this method could provide a new possibility for the development of an order-made therapeutic vaccine against HIV-1.
AB - Development of a safe and effective vaccine against human immunodeficiency virus (HIV) is urgent, but many concerns regarding the safety and efficacy of the currently developing vaccines remain. A major hindrance in HIV vaccine development is the genetic diversity, a hallmark of HIV biology, and a poor understanding of how HIV vaccine prevents the emergence of escape variants during infection and progression of AIDS. Here, we developed a method to construct a molecularly cloned viral library. This technique employs a long-range polymerase chain reaction (PCR) to amplify a virtually full-length HIV type 1 (HIV-1) provirus genome from peripheral blood mononuclear cells (PBMCs) infected with CRF01_AE (subtype E) Thai primary isolate. Among randomly selected 93 clones, 41 with a full-length sequence were able to replicate in PBMCs, 5 of which induced strong cytopathic effects. Replication kinetics also showed that the parental virus was intermediate among the clones. Thus, the molecular library prepared by this method showed the quasi-species in infected cells and this method could provide a new possibility for the development of an order-made therapeutic vaccine against HIV-1.
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U2 - 10.1016/S0264-410X(01)00429-7
DO - 10.1016/S0264-410X(01)00429-7
M3 - Article
C2 - 11803080
AN - SCOPUS:0037081385
SN - 0264-410X
VL - 20
SP - 1181
EP - 1185
JO - Vaccine
JF - Vaccine
IS - 7-8
ER -
