scholarly journals γδ-TCR+ CD2– lymphocytes are recruited into bovine mammary gland after stimulation

2012 ◽  
Vol 51 (No. 5) ◽  
pp. 258-264 ◽  
Author(s):  
M. Faldyna ◽  
L. Leva ◽  
Z. Sladek ◽  
D. Rysanek ◽  
M. Toman
Keyword(s):  
Flow Cytometry ◽  
Mammary Gland ◽  
T Cell ◽  
T Cell Receptor ◽  
Muramyl Dipeptide ◽  
Cell Receptor ◽  
Bovine Mammary Gland ◽  
Gamma Delta ◽  
Colour Flow ◽  
Before And After

&gamma;&delta;-T-Cell Receptor (TCR) lymphocytes were detected in mammary gland lavages collected from 10 clinically healthy virgin heifers before and after intramammary stimulation with synthetic muramyl dipeptide analogue. Using two-colour flow cytometry, CD2<sup>+</sup> and CD2<sup>&ndash;</sup> subsets of &gamma;&delta;-TCR lymphocytes were analyzed. CD2<sup>+</sup> &gamma;&delta;-TCR lymphocytes markedly prevailed over CD2<sup>&ndash;</sup> cells in intact mammary gland: 88.9 &plusmn; 4.9% of &gamma;&delta;-TCR lymphocytes were CD2<sup>+</sup>. After stimulation, neutrophils and &gamma;&delta;-TCR lymphocytes were recruited into the mammary gland. Among &gamma;&delta;-TCR lymphocytes, CD2<sup>&ndash;</sup> cells were mainly responsible for their expansion. After stimulation, 60.8 &plusmn; 13.4% of &gamma;&delta;-TCR lymphocytes were CD2<sup>+</sup> (P &lt; 0.01 when compared with mammary gland lavages before stimulation). It follows from the present study that the cells seem to be involved in the first phase of a response to an infection affecting mammary gland.

scholarly journals A Multiplex, Droplet Digital PCR Assay for the Detection of T-Cell Receptor Excision Circles and Kappa-Deleting Recombination Excision Circles

2019 ◽  
Vol 66 (1) ◽  
pp. 229-238 ◽  
Author(s):  
Tracie Profaizer ◽  
Patricia Slev
Keyword(s):  
Flow Cytometry ◽  
T Cell ◽  
T Cell Receptor ◽  
Reference Gene ◽  
Cell Receptor ◽  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Blood Samples ◽  
Healthy Donors ◽  
Excision Circles

Abstract BACKGROUND T-cell receptor excision circles (TREC) and κ-deleting recombination receptor excision circles (KREC) concentrations can be used to assess and diagnose immune deficiencies, monitor thymic and bone marrow immune reconstitution, or follow responses to drug therapy. We developed an assay to quantify TREC, KREC, and a reference gene in a single reaction using droplet digital PCR (ddPCR). METHODS PCR was optimized for 3 targets: TREC, KREC, and ribonuclease P/MRP subunit p30 (RPP30) as the reference gene. Multiplexing was accomplished by varying the target's fluorophore and concentration. Correlation with clinical results was evaluated using 47 samples from healthy donors, 59 samples with T-cell and B-cell markers within the reference interval from the flow cytometry laboratory, 20 cord blood samples, and 34 samples submitted for exome sequencing for severe combined immunodeficiency disease (SCID). RESULTS The limit of the blank was 4 positive droplets, limit of detection 9 positive droplets, and limit of quantification 25 positive droplets, or 2.0 copies/μL. TREC and KREC copies/μL were as expected in the healthy donors and cord blood samples and concordant with the healthy flow cytometry results. Of the samples from the SCID Panel, 56.5% had a TREC count &lt;20 copies/μL and 17.7% had a KREC count &lt;20 copies/μL, suggestive of low T- and B-cell numbers, respectively. CONCLUSIONS Our multiplex ddPCR assay is an analytically sensitive and specific method for the absolute quantification of TREC and KREC. To the best of our knowledge, this paper is the first to describe the simultaneous quantification of TREC, KREC, and a reference gene by use of ddPCR.

scholarly journals Amino acid substitutions in the floor of the putative antigen-binding site of H-2T22 affect recognition by a gamma delta T-cell receptor

1993 ◽  
Vol 90 (23) ◽  
pp. 11396-11400 ◽  
Author(s):  
S Moriwaki ◽  
B S Korn ◽  
Y Ichikawa ◽  
L van Kaer ◽  
S Tonegawa
Keyword(s):  
T Cell ◽  
T Cell Receptor ◽  
Cell Receptor ◽  
Peptide Transporter ◽  
Affect Recognition ◽  
Antigen Binding ◽  
In Vitro Mutagenesis ◽  
Gamma Delta ◽  
T Cell Clone ◽  
Gamma Delta T Cell

We have previously identified a self-reactive gamma delta T-cell clone (KN6) specific for the H-2T region gene product T22b. Now we have investigated by an in vitro mutagenesis analysis of the T22b gene the possibility that the interaction between the KN6 gamma delta T-cell receptor and T22b involves a peptide. The results demonstrate that mutations at the floor of the putative antigen-binding groove of T22b affect recognition by the gamma delta T-cell receptor. Furthermore, we have shown that KN6 cells react with cells that are deficient in the class I peptide transporter TAP1/TAP2. These results suggest that peptide is involved in the interaction of the KN6 T-cell receptor with T22 and that loading of T22 with the putative peptide is TAP1/TAP2-independent.

scholarly journals Regulated expression and structure of T cell receptor gamma/delta transcripts in human thymic ontogeny.

1991 ◽  
Vol 10 (1) ◽  
pp. 83-91 ◽  
Author(s):  
L. D. McVay ◽  
S. R. Carding ◽  
K. Bottomly ◽  
A. C. Hayday
Keyword(s):  
T Cell ◽  
T Cell Receptor ◽  
Cell Receptor ◽  
Gamma Delta ◽  
Regulated Expression

scholarly journals Gamma/delta lineage relationship within a consecutive series of human precursor T-cell neoplasms

Blood ◽  
1989 ◽  
Vol 74 (7) ◽  
pp. 2508-2518 ◽  
Author(s):  
JP de Villartay ◽  
AB Pullman ◽  
R Andrade ◽  
E Tschachler ◽  
O Colamenici ◽  
...  
Keyword(s):  
T Cell ◽  
T Cell Receptor ◽  
Cell Receptor ◽  
Consecutive Series ◽  
Gene Rearrangements ◽  
Gamma Delta ◽  
Delta Gene ◽  
Gene Usage ◽  
Rearrangement Process ◽  
Gamma Delta T Cell

Abstract We analyzed the gene rearrangements associated with the newly described delta T-cell receptor (TCR) gene from a series of 19 consecutive precursor T-cell (lymphoblastic) neoplasms that represent discrete stages surrounding the TCR gene rearrangement process. Significantly, the delta TCR gene showed rearrangement in most (13 of 19) of these T cells, and in addition it was rearranged in two cells displaying no rearrangement for any other TCR gene. Our survey showed three types of delta gene rearrangements associated with cell-surface TCR expression that presumably represent usage of three V delta genes. This analysis demonstrates (1) a major subclass of human precursor T-cell neoplasms belonging to the gamma/delta T-cell receptor-rearranging subtype; (2) a narrow repertoire of human V delta gene usage; and (3) the utility of delta gene rearrangements as a diagnostic clonal marker in precursor T lymphoblastic neoplasms.

scholarly journals A role for interleukin 4 in the differentiation of mature T cell receptor gamma/delta + cells from human intrathymic T cell precursors.

1990 ◽  
Vol 172 (2) ◽  
pp. 439-446 ◽  
Author(s):  
A Bárcena ◽  
M L Toribio ◽  
L Pezzi ◽  
C Martínez
Keyword(s):  
T Cells ◽  
T Cell ◽  
T Cell Receptor ◽  
Critical Role ◽  
Cell Receptor ◽  
Interleukin 4 ◽  
Gamma Delta ◽  
Consistent Finding ◽  
Delta Cells ◽  
Gamma Delta Cells

We have analyzed the effect of human recombinant interleukin 4 (rIL-4) on the growth and differentiation of human intrathymic pre-T cells (CD7+2+1-3-4-8-). We describe that this population of T cell precursors proliferates in response to rIL-4 (in the absence of mitogens or other stimulatory signals) in a dose-dependent way. The IL-4-induced proliferation is independent of the IL-2 pathway, as it cannot be inhibited with an anti-IL-2 receptor alpha chain antibody. In our culture conditions, rIL-4 also promotes the differentiation of pre-T cells into phenotypically mature T cells. Although both CD3/T cell receptor (TCR)-alpha/beta + and CD3-gamma/delta + T cells were obtained, the preferential differentiation into TCR-gamma/delta + cells was a consistent finding. These results suggest that, in addition to IL-2, IL-4 plays a critical role in promoting growth and differentiation of intrathymic T cell precursors at early stages of T cell development.

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