Human T Cell Priming Assay: Depletion of Peripheral Blood Lymphocytes in CD25+ Cells Improves the In Vitro Detection of Weak Allergen-Specific T Cells

2013 ◽  
pp. 89-100 ◽  
Author(s):  
Marc Vocanson ◽  
Amine Achachi ◽  
Virginie Mutez ◽  
Magalie Cluzel-Tailhardat ◽  
Béatrice Le Varlet ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Peripheral Blood ◽  
Peripheral Blood Lymphocytes ◽  
Blood Lymphocytes ◽  
Human T Cell ◽  
T Cell Priming

scholarly journals Expansion of a lymphocyte population co-expressing T4 (CD4) and T8 (CD8) antigens in the peripheral blood of a normal adult male

Blood ◽  
1990 ◽  
Vol 75 (10) ◽  
pp. 2024-2029 ◽  
Author(s):  
NE Kay ◽  
N Bone ◽  
M Hupke ◽  
AP Dalmasso
Keyword(s):  
T Cells ◽  
T Cell ◽  
B Cell ◽  
Adult Male ◽  
Peripheral Blood ◽  
Peripheral Blood Lymphocytes ◽  
Normal Adult ◽  
Cd8 Cells ◽  
Blood Lymphocytes

Approximately 2% to 3% of circulating human T cells co-express CD4 and CD8 (CD4+, CD8+) T-cell antigens. These CD4+, CD8+ cells may be immature precursors that function to replenish functional T-cell subsets. We detected a very high level of CD4+, CD8+ cells in the peripheral blood lymphocytes of a healthy white male, ranging from 21% to 36%. The morphology of his peripheral blood lymphocytes was normal, and he has maintained an elevated level of CD4+, CD8+ cells without clinical disease over a 19-month observation period. The CD4+, CD8+ cells did not possess thymocyte membrane antigen (CD6), nor did they have increased Tac (CD25) antigen. His intact, purified blood T cells had a normal proliferative response to phytohemagglutinin and interleukin-2 (IL-2), provided help for B-cell proliferation at control levels and exposure to IL-2 resulted in generation of cytotoxic cells. However, the purified blood CD4+, CD8+ cells were deficient in these latter functions except for help in B-cell function. Despite defective function, the isolated CD4+, CD8+ cells co-expressed CD2 and CD3. Prolonged in vitro culture of CD4+, CD8+ cells was possible in the presence of recombinant IL-2. The cultured CD4+, CD8+ cells retained the double antigens (CD4 and CD8) throughout a 4-week period. It is likely these cells are less mature than CD4+, CD8- or CD4-, CD8+ T cells as the CD4+, CD8+ have less in vitro function than the former cells, but it is not yet clear if they mature into either CD4+, CD8-, or CD4-, CD8+ cells. Finally, the presence of an expanded, hypofunctioning CD4+, CD8+ cell population in a normal adult male is apparently compatible with excellent clinical health.

scholarly journals Expansion of a lymphocyte population co-expressing T4 (CD4) and T8 (CD8) antigens in the peripheral blood of a normal adult male

Blood ◽  
1990 ◽  
Vol 75 (10) ◽  
pp. 2024-2029 ◽  
Author(s):  
NE Kay ◽  
N Bone ◽  
M Hupke ◽  
AP Dalmasso
Keyword(s):  
T Cells ◽  
T Cell ◽  
B Cell ◽  
Adult Male ◽  
Peripheral Blood ◽  
Peripheral Blood Lymphocytes ◽  
Normal Adult ◽  
Cd8 Cells ◽  
Blood Lymphocytes

Abstract Approximately 2% to 3% of circulating human T cells co-express CD4 and CD8 (CD4+, CD8+) T-cell antigens. These CD4+, CD8+ cells may be immature precursors that function to replenish functional T-cell subsets. We detected a very high level of CD4+, CD8+ cells in the peripheral blood lymphocytes of a healthy white male, ranging from 21% to 36%. The morphology of his peripheral blood lymphocytes was normal, and he has maintained an elevated level of CD4+, CD8+ cells without clinical disease over a 19-month observation period. The CD4+, CD8+ cells did not possess thymocyte membrane antigen (CD6), nor did they have increased Tac (CD25) antigen. His intact, purified blood T cells had a normal proliferative response to phytohemagglutinin and interleukin-2 (IL-2), provided help for B-cell proliferation at control levels and exposure to IL-2 resulted in generation of cytotoxic cells. However, the purified blood CD4+, CD8+ cells were deficient in these latter functions except for help in B-cell function. Despite defective function, the isolated CD4+, CD8+ cells co-expressed CD2 and CD3. Prolonged in vitro culture of CD4+, CD8+ cells was possible in the presence of recombinant IL-2. The cultured CD4+, CD8+ cells retained the double antigens (CD4 and CD8) throughout a 4-week period. It is likely these cells are less mature than CD4+, CD8- or CD4-, CD8+ T cells as the CD4+, CD8+ have less in vitro function than the former cells, but it is not yet clear if they mature into either CD4+, CD8-, or CD4-, CD8+ cells. Finally, the presence of an expanded, hypofunctioning CD4+, CD8+ cell population in a normal adult male is apparently compatible with excellent clinical health.

T cell receptor Vb5 and Vb17 clonal diversity in cerebrospinal fluid and peripheral blood lymphocytes of multiple sclerosis patients

1998 ◽  
Vol 4 (3) ◽  
pp. 154-161 ◽  
Author(s):  
P Lozeron ◽  
D Chabas ◽  
B Duprey ◽  
O Lyon-Caen ◽  
R Liblau
Keyword(s):  
Central Nervous System ◽  
T Cells ◽  
Nervous System ◽  
T Cell ◽  
T Cell Receptor ◽  
Peripheral Blood ◽  
Cell Receptor ◽  
Peripheral Blood Lymphocytes ◽  
Blood Lymphocytes ◽  
Multiple Sclerosis Patients

To better characterize the cellular immune response taking place in the MS central nervous system, we investigated the blood and CSF T cell receptor (TCR) Vβ5 and Vb17 repertoire in HLA-typed patients with recently diagnosed MS or other neurological diseases (OND). Using a RT-PCR based technique, we analysed directly ex vivo the CDR3 size of TCR β chains utilizing Vβ5 (eight patients with MS and one with OND) or Vβ17 (eight patients with MS and six with OND) gene segments on paired blood-CSF samples. Globally, the analysis of Vβ5-Jβ and Vβ17-Jβ repertoire showed a less diverse pattern in the CSF samples than in the corresponding peripheral blood lymphocytes both in MS and in OND patients. However, we did not detect any recurrent clonal expansion within the Vb5+ T cells in MS patients, underlining the potential limits of Vβ5- based immunotherapy in MS. We found an expanded T cell population using the same Vβ17-Jβ1.6 combination with identical CDR3 length in the CSF of three MS patients and none of the control patients. These results suggest selective expansion of T cells expressing this segment gene in the MS central nervous system.

scholarly journals Activation and increased expression of adhesion molecules on peripheral blood lymphocytes is a mechanism for the immediate lymphocytopenia after administration of OKT3

Blood ◽  
1996 ◽  
Vol 87 (1) ◽  
pp. 404-411 ◽  
Author(s):  
S Buysmann ◽  
FJ Bemelman ◽  
PT Schellekens ◽  
Y van Kooyk ◽  
CG Figdor ◽  
...  
Keyword(s):  
T Cells ◽  
Monoclonal Antibodies ◽  
T Lymphocytes ◽  
Adhesion Molecules ◽  
Peripheral Blood ◽  
Peripheral Blood Lymphocytes ◽  
Intercellular Adhesion Molecule ◽  
Intercellular Adhesion Molecule 1 ◽  
Blood Lymphocytes

Abstract We investigated the mechanism by which antihuman CD3 monoclonal antibodies of the isotypes IgG2a (eg, OKT3) and IgA (eg, IXA) can induce the rapid disappearance of virtually all circulating T lymphocytes. We hypothesize that upregulation of adhesion molecules on the lymphocyte membrane contributes to this effect. However, this hypothesis is difficult to test, because of the inherent lymphocytopenia and/or shifts in lymphocyte populations between intra and extra-vascular compartments. Therefore, studies in vitro were performed, as well. Analysis of peripheral blood lymphocytes isolated at several times after addition of OKT3 or IXA to whole blood of healthy individuals showed an immediate increase in the proportion of T cells expressing NKI-L16, an activation epitope on CD11a/CD18. Likewise, an increase in CD11b/CD18 expression occurred. In parallel experiments, a transiently increased adhesion of T cells to endothelial cell monolayers was observed. This adhesion could be completely blocked by anti-CD18 or anti-CD11a monoclonal antibodies and only partly by an anti-CD11b antibody. Our data indicate that upregulation of activation epitopes of CD11a/CD18, as well as increased expression of CD11b/CD18 on T lymphocytes, may result in increased adhesion of these cells to intercellular adhesion molecule-1 (ICAM-1) and ICAM-2 on vascular endothelium. This phenomenon may, at least, partly explain the rapidly occurring peripheral lymphocytopenia observed in vivo.

Immunological functions and phenotypes of peripheral blood lymphocytes from human T-Cell leukemia virus-I carriers

1989 ◽  
Vol 9 (6) ◽  
pp. 477-484 ◽  
Author(s):  
Yoshiya Tanaka ◽  
Susumu Oda ◽  
Kazuhiko Nagata ◽  
Naoki Mori ◽  
Hisahiro Sakamoto ◽  
...  
Keyword(s):  
T Cell ◽  
Peripheral Blood ◽  
Leukemia Virus ◽  
Peripheral Blood Lymphocytes ◽  
T Cell Leukemia ◽  
Cell Leukemia ◽  
Blood Lymphocytes ◽  
Human T Cell ◽  
T Cell Leukemia Virus

scholarly journals Identification and physical mapping of a polymorphic human T cell receptor V beta gene with a frequent null allele.

1993 ◽  
Vol 177 (1) ◽  
pp. 135-143 ◽  
Author(s):  
P Charmley ◽  
K Wang ◽  
L Hood ◽  
D A Nickerson
Keyword(s):  
T Cell ◽  
Peripheral Blood ◽  
Null Allele ◽  
Gene Segment ◽  
Peripheral Blood Lymphocytes ◽  
Dna Polymorphisms ◽  
Blood Lymphocytes ◽  
Human T Cell ◽  
Beta 2 ◽  
Beta Gene

Germline variation in genes that encode the human T cell receptors (TCRs) may have an important influence in shaping the immune T cell repertoire. In this report we describe a frequent null allele of the human V beta 18 gene, resulting from a nucleotide substitution that creates a stop codon (CGA<-->TGA). Approximately 11% of the population tested was homozygous for this null allele, indicating that this is a frequent "hole in the repertoire." We confirmed that there is a greatly reduced (undetectable) level of V beta 18 mRNA in peripheral blood lymphocytes from an individual homozygous for this null allele. In addition, all heterozygous individuals expressed detectable levels of only the functional V beta 18 allele in their peripheral blood lymphocytes. Two other DNA polymorphisms were identified in V beta 18, one of which would result in an amino acid substitution in an expressed V beta 18 gene. Genotypes for all three of these V beta 18 DNA polymorphisms were determined in a group of unrelated individuals. Statistical analyses of the associations between alleles of the V beta 18 polymorphisms and those of other DNA polymorphisms in the TCR beta locus suggested a close physical proximity between the V beta 18 gene and the 3' end of the C beta 2 region. This localization of human V beta 18 had been previously predicted by the sequence homology between human V beta 18 and mouse V beta 14, a V gene segment previously mapped to 3' of the mouse C beta genes. We confirmed this localization of the human V beta 18 gene by isolating a cosmid clone that contains both the V beta 18 and C beta 2 segments. Mapping by restriction enzyme digestion and by the polymerase chain reaction indicated that the V beta 18 gene segment is approximately 9 kb 3' of the C beta 2 gene, making this the only known human V beta gene 3' of the C beta region.

scholarly journals Activation and increased expression of adhesion molecules on peripheral blood lymphocytes is a mechanism for the immediate lymphocytopenia after administration of OKT3

Blood ◽  
1996 ◽  
Vol 87 (1) ◽  
pp. 404-411 ◽  
Author(s):  
S Buysmann ◽  
FJ Bemelman ◽  
PT Schellekens ◽  
Y van Kooyk ◽  
CG Figdor ◽  
...  
Keyword(s):  
T Cells ◽  
Monoclonal Antibodies ◽  
T Lymphocytes ◽  
Adhesion Molecules ◽  
Peripheral Blood ◽  
Peripheral Blood Lymphocytes ◽  
Intercellular Adhesion Molecule ◽  
Intercellular Adhesion Molecule 1 ◽  
Blood Lymphocytes

We investigated the mechanism by which antihuman CD3 monoclonal antibodies of the isotypes IgG2a (eg, OKT3) and IgA (eg, IXA) can induce the rapid disappearance of virtually all circulating T lymphocytes. We hypothesize that upregulation of adhesion molecules on the lymphocyte membrane contributes to this effect. However, this hypothesis is difficult to test, because of the inherent lymphocytopenia and/or shifts in lymphocyte populations between intra and extra-vascular compartments. Therefore, studies in vitro were performed, as well. Analysis of peripheral blood lymphocytes isolated at several times after addition of OKT3 or IXA to whole blood of healthy individuals showed an immediate increase in the proportion of T cells expressing NKI-L16, an activation epitope on CD11a/CD18. Likewise, an increase in CD11b/CD18 expression occurred. In parallel experiments, a transiently increased adhesion of T cells to endothelial cell monolayers was observed. This adhesion could be completely blocked by anti-CD18 or anti-CD11a monoclonal antibodies and only partly by an anti-CD11b antibody. Our data indicate that upregulation of activation epitopes of CD11a/CD18, as well as increased expression of CD11b/CD18 on T lymphocytes, may result in increased adhesion of these cells to intercellular adhesion molecule-1 (ICAM-1) and ICAM-2 on vascular endothelium. This phenomenon may, at least, partly explain the rapidly occurring peripheral lymphocytopenia observed in vivo.

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