PMID- 12191511
OWN - NLM
STAT- MEDLINE
DCOM- 20020903
LR  - 20191210
IS  - 0022-1759 (Print)
IS  - 0022-1759 (Linking)
VI  - 264
IP  - 1-2
DP  - 2002 Jun 1
TI  - Development of a novel allo-independent HIV-1 virus preparation for use in 
      immunoassays.
PG  - 69-76
AB  - The study of the immunologic response to whole human immunodeficiency virus type 
      1 (HIV-1) antigen is limited by the presence of highly immunogenic human 
      leukocyte antigen (HLA) alloantigens on the envelope of wild type virus. This 
      paper outlines the production of HIV-1 infectious virions free of HLA for use as 
      whole viral antigens in immunoassays. An infectious molecular clone of HIV-1 was 
      transfected into the K-562 cell line, which does not express HLA on the cell 
      surface. After a 30-day selection period, to ensure stable transfection, cells 
      and culture supernatants were analyzed for productive HIV-1 infection and virion 
      infectivity. An enzyme-linked immunosorbent assay (ELISA) confirmed the presence 
      of p24 in the culture supernatants. Molecular confirmation of HIV-1 transfection 
      was achieved by gene amplification. Flow cytometric analysis was used to identify 
      gp120 on the surface of the infected cells. Viral supernatants were tested for 
      HIV infectivity in peripheral blood mononuclear cells (PBMCs). The usefulness of 
      this viral preparation as whole virus antigens was validated using PBMCs from 
      HIV-infected individuals. These results indicate the successful production of 
      HIV-1 infectious virions, which do not have HLA molecules on their viral 
      envelope, and demonstrate their utility for immunoassays.
FAU - Koesters, Sandra
AU  - Koesters S
AD  - Department of Medical Microbiology, University of Manitoba, 539-730 William 
      Avenue, Winnipeg, MB, Canada.
FAU - Rutherford, W
AU  - Rutherford W
FAU - Fowke, Keith R
AU  - Fowke KR
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Validation Study
PL  - Netherlands
TA  - J Immunol Methods
JT  - Journal of immunological methods
JID - 1305440
RN  - 0 (Gene Products, nef)
RN  - 0 (HIV Antigens)
RN  - 0 (HIV Core Protein p24)
RN  - 0 (HIV Envelope Protein gp120)
RN  - 0 (HLA Antigens)
RN  - 0 (HLA-DR Antigens)
RN  - 0 (Histocompatibility Antigens Class I)
RN  - 0 (nef Gene Products, Human Immunodeficiency Virus)
SB  - IM
MH  - Cell Culture Techniques/methods
MH  - Cell-Free System/immunology
MH  - Cells, Cultured
MH  - Gene Products, nef/biosynthesis/genetics
MH  - HIV Antigens/genetics/*immunology
MH  - HIV Core Protein p24/biosynthesis/genetics
MH  - HIV Envelope Protein gp120/biosynthesis/genetics
MH  - HIV Infections/immunology
MH  - HIV-1/genetics/*immunology
MH  - HLA Antigens/biosynthesis/genetics/*physiology
MH  - HLA-DR Antigens/biosynthesis/genetics
MH  - Histocompatibility Antigens Class I/biosynthesis/genetics
MH  - Humans
MH  - Immunoassay/*methods
MH  - K562 Cells/immunology/virology
MH  - Transfection
MH  - Transgenes
MH  - nef Gene Products, Human Immunodeficiency Virus
EDAT- 2002/08/23 10:00
MHDA- 2002/09/11 10:01
CRDT- 2002/08/23 10:00
PHST- 2002/08/23 10:00 [pubmed]
PHST- 2002/09/11 10:01 [medline]
PHST- 2002/08/23 10:00 [entrez]
AID - S0022175902000856 [pii]
AID - 10.1016/s0022-1759(02)00085-6 [doi]
PST - ppublish
SO  - J Immunol Methods. 2002 Jun 1;264(1-2):69-76. doi: 10.1016/s0022-1759(02)00085-6.