PMID- 14594509
OWN - NLM
STAT- MEDLINE
DCOM- 20040609
LR  - 20171116
IS  - 1525-8165 (Print)
IS  - 1525-8165 (Linking)
VI  - 12
IP  - 5
DP  - 2003 Oct
TI  - Functional evaluation of proliferative T cell responses in patients with severe T 
      lymphopenia: characterization of optimal culture conditions and standardized 
      activation signals for a simple whole blood assay.
PG  - 525-35
AB  - In this methodological study, we describe an assay for analysis of proliferative 
      T cell responses in patients with severe leukopenia. Severe treatment-induced 
      cytopenia is observed in patients with malignant disorders who receive 
      conventional intensive chemotherapy or autologous stem cell transplantation. The 
      quantitative T cell defect can then be characterized by flow cytometric analysis 
      of membrane molecule expression, whereas the functional status of the remaining T 
      cell population is more difficult to evaluate. In the present study, we describe 
      a standardized whole blood assay that requires small sample volumes and can be 
      used for repeated analysis even in severely ill patients. The assay is based on 
      the following strategy: (i) blood samples are diluted with the serum-free medium 
      X-vivo 10, (ii) T cells are activated either with monoclonal immunoglobulin E 
      (IgE) anti-CD3 or anti-CD3 plus anti-CD28; (iii) T cell proliferation is assayed 
      by [(3)H]thymidine incorporation after 4 days of in vitro culture. These 
      proliferative responses are not affected by the plasma levels of interleukin-2 
      (IL-2), sIL-2-R alpha, IL-7 and IL-15, and the kinetics of the response are not 
      altered by the presence of exogenous cytokines. Detectable proliferation is 
      observed for most patients with treatment-induced cytopenia. We conclude that the 
      assay can be used for functional characterization of remaining T lymphocytes in 
      patients with severe T lymphopenia.
FAU - Wendelbo, Oystein
AU  - Wendelbo O
AD  - Division for Hematology, Department of Medicine, Haukeland University Hospital, 
      Bergen, Norway. wend@haukeland.no
FAU - Bruserud, Oystein
AU  - Bruserud O
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Hematother Stem Cell Res
JT  - Journal of hematotherapy & stem cell research
JID - 100892915
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Antigens, CD)
RN  - 0 (CD28 Antigens)
RN  - 0 (CD3 Complex)
RN  - 0 (Culture Media)
RN  - 0 (Interleukins)
RN  - 0 (Proto-Oncogene Proteins)
RN  - 83869-56-1 (Granulocyte-Macrophage Colony-Stimulating Factor)
RN  - EC 2.7.10.1 (FLT3 protein, human)
RN  - EC 2.7.10.1 (Receptor Protein-Tyrosine Kinases)
RN  - EC 2.7.10.1 (fms-Like Tyrosine Kinase 3)
SB  - IM
MH  - Adolescent
MH  - Adult
MH  - Aged
MH  - Antibodies, Monoclonal/pharmacology
MH  - Antigens, CD/analysis
MH  - CD28 Antigens/immunology
MH  - CD3 Complex/immunology
MH  - Cell Culture Techniques/methods
MH  - Cell Division/drug effects
MH  - Culture Media/pharmacology
MH  - Drug-Related Side Effects and Adverse Reactions
MH  - Female
MH  - Flow Cytometry
MH  - Granulocyte-Macrophage Colony-Stimulating Factor/pharmacology
MH  - Humans
MH  - Interleukins/blood/pharmacology
MH  - Leukemia/blood/therapy
MH  - Leukocytes, Mononuclear/chemistry/metabolism
MH  - Lymphocyte Activation/drug effects/*immunology
MH  - Lymphopenia/*blood/etiology/immunology
MH  - Male
MH  - Middle Aged
MH  - Multiple Myeloma/blood/therapy
MH  - Myelodysplastic Syndromes/blood/therapy
MH  - Proto-Oncogene Proteins/pharmacology
MH  - Receptor Protein-Tyrosine Kinases/pharmacology
MH  - Stem Cell Transplantation/adverse effects
MH  - T-Lymphocytes/cytology/drug effects/*metabolism
MH  - Time Factors
MH  - fms-Like Tyrosine Kinase 3
EDAT- 2003/11/05 05:00
MHDA- 2004/06/21 10:00
CRDT- 2003/11/05 05:00
PHST- 2003/11/05 05:00 [pubmed]
PHST- 2004/06/21 10:00 [medline]
PHST- 2003/11/05 05:00 [entrez]
AID - 10.1089/152581603322448231 [doi]
PST - ppublish
SO  - J Hematother Stem Cell Res. 2003 Oct;12(5):525-35. doi: 
      10.1089/152581603322448231.