Expression Profile and Clonality of T‐Cell Receptor Beta Variable Genes in Normal Human Endometrium

Abstract

Our results stress the similarities between T cells residing in different mucosal tissues and provide a basis for future investigations about endometrial T cells and their antigen specificities in gynecological problems.

HLA-Cw and TCR vβ analysis in twenty Mexican patients with psoriasis

“…Statistical analysis, including transcript and protein results, suggests two major restricted T-cell expansions: TCR Vβ-2 and TCR Vβ-7. Evidence of the TCR Vβ-2 expression bias has been already reported for T-cells that are destined to migrate to the skin [18], and TCR Vβ-7 expression has been studied in psoriasis, as well as in normal tissues and other diseases [9,19]. Since the majority of the samples from Group 1 were positive to TCRs using Western blot in PBMC, we decided to examine random skin biopsies Group (n=6) by immunohistochemistry.…”

ORIGINAL ARTICLE: Repertoire and Clonality of T‐Cell Receptor Beta Variable Genes Expressed in Endometrium and Blood T Cells of Patients with Recurrent Spontaneous Abortion

“…Molecular procedures used in this study have been fully described before 15 . Briefly, total RNA was extracted from PBMC or endometrial tissues after complete homogenization using RNABee (Biogene, Kimbolton, UK) and based on the guanidine thiocyanate phenol–chloroform extraction method.…”

“…The used cDNA concentration in all samples was normalized using a TCR beta chain constant region (BC)‐specific polymerase chain reaction (TCRBC PCR) by a set of forward (5′‐BC) and reverse (3′‐BC) primers specific for the constant region of the beta chain. 15 Approximately equal amounts of cDNA were used in each TCRBV‐BC PCR according to the quantified density of different cDNA concentrations obtained by densitometry of electrophoresed bands of TCRBC PCR products.…”

“…Analysis of BV‐gene usage was carried out by amplification of BV‐specific cDNA using a panel of 28 TCR BV‐specific primers as previously described 15 . PCR mixtures for blood samples contained 1 unit of Taq DNA polymerase (Cinnagen, Tehran, Iran) in its recommended 1x buffer (10 m m Tris–HCl, 50 mM KCl), 0.2 m m final concentration of each dNTPs, 1.5 m m MgCl 2 and 0.6 μ m of each primer in the final volume of 25 μL for each reaction.…”

“…Membranes were air‐dried and baked for 2 hr at 80°C and stored for subsequent steps. A TCR BC‐specific oligonucleotide probe (BC reporter), recognizing both BC1 and BC2 upstream of the BC primer 15 was used and was 5′‐end labeled using T4 polynucleotide kinase enzyme (Invitrogen, Paisley, UK) and γ‐ 32 P‐adenosine triphosphate (GE Healthcare). The baked nylon membranes were first pre‐hybridized in a buffer containing 2x SSPE, 5x Denhardt’s solution, and 0.5% sodium dodecyl sulfate (SDS) for 3 hr at 42°C, followed by hybridization in the same buffer with 1 × 10 6 c.p.m./mL of the 5′‐end‐labeled BC reporter overnight at 42°C.…”