Current status and recent advances of next generation sequencing techniques in immunological repertoire.

To ward off a wide variety of pathogens, the human adaptive immune system harbors a vast array of T-cell receptors (TCRs) and B-cell receptors (BCRs), collectively referred to as the immune repertoire. High-throughput sequencing (HTS) of TCR/BCR genes allows in-depth molecular analysis of T/B-cell clones, providing an unprecedented level of detail when examining the T/B-cell repertoire of individuals. It can evaluate TCR/BCR complementarity-determining region 3 (CDR3) diversity and assess the clonal composition, including the size of the repertoire; similarities between repertoires; V(D)J segment use; nucleotide insertions and deletions; CDR3 lengths; and amino acid distributions along the CDR3s at sequence-level resolution. Deep sequencing of B-cell and T-cell repertoires offers the potential for a quantitative understanding of the adaptive immune system in healthy and disease states. Recently, paired sequencing strategies have also been developed, which can provide information about the identity of immune receptor pairs encoded by individual T or B lymphocytes. HTS technology provides a previously unimaginable amount of sequence data, accompanied, however, by numerous challenges associated with error correction and interpretation that remain to be solved. The review details some of the technologies and some of the recent achievements in this field.

Study of a novel coating strategy for coronary stents: evaluation of stainless metallic steel coated with VEGF and anti-CD34 antibody in vitro.

OBJECTIVE: To access the cytotoxicity and the effect on the endothelial progenitor cell (EPC) differentiation of stainless steel sheets simultaneously coated with VEGF and anti-CD34 antibody. MATERIALS AND METHODS: 316L stainless steel sheets (diameter 6 mm, thickness 1 mm) were divided into the D-H (Bare metal), D-(H-V)10 (VEGF-coated metal) and D-(H-V)10-A (VEGF and anti-CD34 antibody co-coated metal) groups. The cytotoxicity effect of the three groups was measured using MTT assay. Percentage of EPC positive for CD34, CD133 and KDR were detected by flow cytometric assay. Endothelial cells positive for CD31 and VE-Cadherin were also detected by flow cytometric assay. RESULTS: The percentages of isolated cells positive for CD133, CD34 and KDR were 89.9%, 91.3%, and 90.4%, respectively, suggesting that the EPCs were successfully isolated. MTT results showed that the stainless steel sheets coated with VEGF and anti-CD34 antibody have less toxicity on seeded EPCs than single VEGF coating or bare metal. We further found that with VEGF and anti-CD34 antibody co-coating could significantly promote the differentiation of EPCs in vitro when compared with that of single VEGF coating and bare metal. CONCLUSIONS: Our study provided a preliminary evaluation of metallic steel sheet coated with VEGF and anti-CD34 antibody in vitro. Our findings suggest that simultaneously coating the stents with VEGF and anti-CD34 antibody might be a novel research direction for facilitating re-endothelialization in order to reduce ISR after stent implantation.

Suppression of late asthmatic response by low-dose oral administration of interferon-beta in the guinea pig model of asthma.

We investigated the anti-asthmatic effects of low-dose oral and subcutaneous administration of interferon-beta (IFN-beta) on an ovalbumin (OVA)-sensitized and challenged guinea pig model of asthma. Subcutaneous administration of IFN-beta suppressed the eosinophil infiltration by 14.2% of the control and the respiratory resistance (Rrs) by 58.2% at 2.0 MIU/kg. Oral administration of IFN-beta inhibited the late asthmatic response (LAR) by suppressing the increase of Rrs by 29% of the control at 1 IU/ml and the eosinophil infiltration into the trachea and lung by 34.6% at the optimum dosage of 10 IU/ml. Both subcutaneous and oral administration could not inhibit the early asthmatic response (EAR). Additionally we found 2',5'-oligoadenylate synthetase (2',5'-OAS) induction by low-dose oral administration (LDOA) of IFN-beta to the same extent as by subcutaneous administration in whole blood in vivo. These data suggest that LDOA of IFN-beta would have some clinical benefit for patients with asthma.