PMID- 15620394
OWN - NLM
STAT- MEDLINE
DCOM- 20050401
LR  - 20081121
IS  - 0166-0934 (Print)
IS  - 0166-0934 (Linking)
VI  - 123
IP  - 2
DP  - 2005 Feb
TI  - Establishment of an alternative intracellular cytokine staining assay for 
      HIV/AIDS clinical studies.
PG  - 131-40
AB  - To evaluate immune responses in large-scale clinical studies, it is crucial to 
      use assays that are not only sensitive and specific, but also standardized and 
      easily transferable. To this end, an intracellular cytokine staining (ICS) assay 
      for quantifying HIV-specific CD8+ T cells has been established and its 
      performance characteristics determined. PBMCs were stimulated using recombinant 
      vaccinia expressing HIV-Env, -Gag, -Pol and -Nef proteins, or a protein from 
      Escherichia coli (Eco); CD8+ T cell responses were assessed by intracellular 
      IFN-gamma staining and flow cytometric (FC) analysis. In 15 HIV seronegative and 
      11 HIV seropositive individuals, a minimal background IFN-gamma staining was 
      found after Eco stimulation with a median [inter-quartile range (IQR)] of 0.005% 
      (0.000%, 0.030%) irrespective of HIV infection status. Recent smallpox 
      vaccination was associated with a small but significant increase in background 
      staining [0.02% (0.02%, 0.04%) versus 0.0% (0.0%, 0.01%) (p=0.039)] in HIV 
      seronegative individuals. This assay detected moderate (>0.10%) HIV-specific 
      responses in 64% (7/11) of HIV seropositive individuals. The results suggest that 
      this ICS assay format is sensitive and specific, and is amenable to 
      standardization for screening for HIV-specific CD8+ T cells in clinical trials.
FAU - Chen, Huiyuan
AU  - Chen H
AD  - Department of Medicine, University of Rochester School of Medicine and Dentistry, 
      Rochester, New York, NY 14642, USA.
FAU - Reichman, Richard
AU  - Reichman R
FAU - Keefer, Michael
AU  - Keefer M
FAU - McDermott, Michael P
AU  - McDermott MP
FAU - Jin, Xia
AU  - Jin X
LA  - eng
GR  - 5MOI-RR00044/RR/NCRR NIH HHS/United States
GR  - R01-AI05163/AI/NIAID NIH HHS/United States
GR  - UO1-AI27658/AI/NIAID NIH HHS/United States
GR  - UO1-AI47980/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - Netherlands
TA  - J Virol Methods
JT  - Journal of virological methods
JID - 8005839
RN  - 0 (Cytokines)
RN  - 82115-62-6 (Interferon-gamma)
SB  - IM
MH  - CD8-Positive T-Lymphocytes/metabolism/virology
MH  - Clinical Trials as Topic
MH  - Cytokines/analysis
MH  - Flow Cytometry
MH  - HIV Infections/*metabolism
MH  - HIV Seropositivity
MH  - Humans
MH  - Interferon-gamma/*analysis
MH  - Staining and Labeling/*methods
MH  - Vaccinia virus/genetics
EDAT- 2004/12/29 09:00
MHDA- 2005/04/02 09:00
CRDT- 2004/12/29 09:00
PHST- 2004/06/12 00:00 [received]
PHST- 2004/09/09 00:00 [revised]
PHST- 2004/09/20 00:00 [accepted]
PHST- 2004/12/29 09:00 [pubmed]
PHST- 2005/04/02 09:00 [medline]
PHST- 2004/12/29 09:00 [entrez]
AID - S0166-0934(04)00280-0 [pii]
AID - 10.1016/j.jviromet.2004.09.013 [doi]
PST - ppublish
SO  - J Virol Methods. 2005 Feb;123(2):131-40. doi: 10.1016/j.jviromet.2004.09.013.