PMID- 9714884
OWN - NLM
STAT- MEDLINE
DCOM- 19981029
LR  - 20201215
IS  - 0736-6205 (Print)
IS  - 0736-6205 (Linking)
VI  - 25
IP  - 2
DP  - 1998 Aug
TI  - Detection and characterization of alpha-beta-T-cell clonality by denaturing 
      gradient gel electrophoresis (DGGE).
PG  - 244-50
AB  - Accumulation of T cells carrying identical T-cell receptors (TCR) is associated 
      with a number of immunological and non-immunological diseases. Therefore, it is 
      of interest to be able to analyze complex T-cell populations for the presence of 
      clonally expanded subpopulations. Here, we describe a simple method combining 
      reverse transcription (RT)-PCR and denaturing gradient gel electrophoresis (DGGE) 
      for rapid detection and characterization of T-cell clonality. The detection of 
      clonally expanded T cells by DGGE relies on the fact that clonal transcripts have 
      no junctional diversity and therefore resolve at a fixed position in the gel, 
      which is determined by their melting properties. For polyclonal populations with 
      a high degree of junctional diversity, the different DNA molecules will resolve 
      at different positions in the gel and together will be revealed as a smear. For 
      each of the TCR beta-variable gene (BV) 1-24 families, cloned transcripts were 
      amplified and shown to resolve as distinct bands in the denaturing gradient gel, 
      whereas the analysis of polyclonal T-cell populations resulted in a smear in the 
      gel. The present method might prove useful to test for clonotypic T-cells in a 
      variety of pathological and physiological conditions and for monitoring T-cell 
      responses in diagnostic and therapeutic settings.
FAU - thor Straten, P
AU  - thor Straten P
AD  - Department of Tumor Cell Biology, Danish Cancer Society, Copenhagen, Denmark.
FAU - Barfoed, A
AU  - Barfoed A
FAU - Seremet, T
AU  - Seremet T
FAU - Saeterdal, I
AU  - Saeterdal I
FAU - Zeuthen, J
AU  - Zeuthen J
FAU - Guldberg, P
AU  - Guldberg P
LA  - eng
PT  - Research Support, Non-U.S. Gov't
PT  - Technical Report
PL  - England
TA  - Biotechniques
JT  - BioTechniques
JID - 8306785
RN  - 0 (DNA Primers)
RN  - 0 (Receptors, Antigen, T-Cell, alpha-beta)
SB  - IM
MH  - Cells, Cultured
MH  - Clone Cells
MH  - DNA Primers
MH  - Electrophoresis, Polyacrylamide Gel/*methods
MH  - Humans
MH  - *Nucleic Acid Denaturation
MH  - Receptors, Antigen, T-Cell, alpha-beta/chemistry/*genetics
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - T-Lymphocytes/*metabolism
MH  - Transcription, Genetic
EDAT- 1998/08/26 00:00
MHDA- 1998/08/26 00:01
CRDT- 1998/08/26 00:00
PHST- 1998/08/26 00:00 [pubmed]
PHST- 1998/08/26 00:01 [medline]
PHST- 1998/08/26 00:00 [entrez]
AID - 10.2144/98252st05 [doi]
PST - ppublish
SO  - Biotechniques. 1998 Aug;25(2):244-50. doi: 10.2144/98252st05.