Modulating the Cellular Immune Response of Oligonucleotides by Brush Polymer-Assisted Compaction.

Unwanted stimulation of the innate immune system by foreign nucleic acids has been one of the major barriers preventing bioactive sequences from reaching market. Foreign nucleic acids can be recognized by multiple pattern recognition receptors (PRRs), which trigger a signaling cascade to activate host defense systems, leading to a range of side effects. This study demonstrates that polyethylene glycol (PEG)-modified DNA strands can greatly reduce the activation of the innate immune system, and the extent of reduction is dependent upon polymer architecture. Highly branched brushes with long PEG side chains achieve the best suppression by blocking PRR interactions via a local steric effect. Interestingly, the brush polymer creates little barrier toward DNA-DNA interaction. Quantification of inflammatory cytokines in both mRNA and protein levels as well as the extent of cellular uptake shows a direct correlation between steric congestion and reduction of cellular immune response. These results suggest that the brush architecture offers unique advantages for PEGylating oligonucleotides in the context of minimizing unwanted immune system activation.

Immunosuppression in specific-pathogen-free broilers administered infectious bursal disease virus vaccines by in ovo route.

The effect of two infectious bursal disease virus (IBDV) vaccines (IBDV-immune complex [Icx] and IBDV-2512), administered in ovo, on the cell-mediated immunity of specific-pathogen-free (SPF) broilers was examined. A decrease (P < 0.05) in the T-cell mitogenic response occurred in birds vaccinated with both vaccines on days 9 and 21 post in ovo vaccination (PIOV), but an increase (P < 0.05) occurred on day 15 PIOV. The T cells from birds given the IBDV-2512 were less responsive. There were no significant differences in proportions of lymphocytes expressing CT4+CT8 and CT8+CT4- except on day 21 PIOV, where an increase (P < 0.05) in IBDV-2512-vaccinated birds and a decrease (P < 0.05) in percentage of CT4+CT8- in IBDV-Icx-vaccinated birds was observed. There was an increase (P < 0.05) in percentage of CT8+CT4- T cells on day 21 PIOV in both vaccinated groups. A decrease (P < 0.05) in B-cell percentage was observed on day 21 PIOV in birds given both vaccines. Results indicated that although humoral immunosuppression is associated with destruction of B cells (bursal atrophy), cell-mediated immunosuppression induced by these two IBDV vaccines in SPF birds was not associated with altered helper (CT4+CT8-) or cytotoxic (CT8+CT4-) subpopulations of T lymphocytes.

Evaluation of the immune response and detection of infectious bursal disease viruses by reverse transcriptase-polymerase chain reaction and enzyme-linked immunosorbent assay after in ovo vaccination of commercial broilers.

Maternal immunity can interfere with infectious bursal disease virus (IBDV) vaccine efficacy. The effect of maternal antibody (MacAb) on the immune response and detection of two vaccine viruses, an IBDV immune complex (cx) and IBDV-2512, was investigated. Vaccines were administered in ovo at 100 mean embryo infectious dose to commerical broiler embryos derived from young (29 wk) or old (63 wk) flocks. Chickens with low MatAb were challenged with an antigenic standard IBDV strain at 21 and 35 days post in ovo vaccination (PIOV). At 28 and 42 days PIOV, birds were euthanatized and bursa weight: body weight ratios were determined. MatAb of progeny from the 29-wk-old flock was higher (P < 0.05) than that from progeny of the 63-wk-old flock. Progeny titers declined by day 21 PIOV. Birds with MatAb vaccinated with either the IBDV-2512 or IBDV-Icx vaccine did not mount an antibody response by 21 days PIOV. The IBDV-Icx vaccination protected chickens against bursal atrophy when they were challenged at 21 and 35 days PIOV (83% and 77%, respectively). Resistance to challenge in the absence of antibody implies that cellular immunity plays a role in IBDV protection. The IBDV-2512 vaccine caused atrophy of the bursae, and, therefore, we could not examine its protection by these criteria. Neither vaccine could be detected by antigen-capture chemiluminescent enzyme-linked immunosorbent assay. Therefore, total RNA was extracted and reverse transcription (RT)-polymerase chain reaction (PCR) and nested PCR were conducted to amplify the VP2 (hypervariable) region on viral RNA collected at 3, 6, 9, 15, and 21 days PIOV. Nested PCR detected the VP2 region of both IBDV vaccine viruses at day 3 PIOV. Only IBDV-2512 RNA was detected on day 6 PIOV. However, the IBDV-Icx RNA was detected on days 9 and 15 PIOV but not on day 21 PIOV. The IBDV-Icx RNA was evident in tissues and could induce protection against challenge. This work gives further insight into the mechanism of the IBDV-Icx vaccine.