Local and Systemic Immunity against Respiratory Syncytial Virus Induced by a Novel Intranasal Vaccine. A Randomized, Double-Blind, Placebo-controlled Clinical Trial

RATIONALE:Needle-free intranasal vaccines offer major potential advantages, especially against pathogens entering via mucosal surfaces. As yet, there is no effective vaccine against respiratory syncytial virus (RSV), a ubiquitous pathogen of global importance that preferentially infects respiratory epithelial cells; new strategies are urgently required. OBJECTIVES:Here, we report the safety and immunogenicity of a novel mucosal RSV F protein vaccine linked to an immunostimulatory bacterium-like particle (BLP). METHODS:In this phase I, randomised, double-blind placebo-controlled trial, 48 healthy volunteers aged 18-49 years were randomly assigned to receive placebo or SynGEM (low- or high-dose) intranasally by prime-boost administration. The primary outcome was safety and tolerability, with secondary objectives assessing virus-specific immunogenicity. MEASUREMENTS AND MAIN RESULTS:There were no significant differences in adverse events between placebo and vaccinated groups. SynGEM induced systemic plasmablast responses and significant, durable increases in RSV-specific serum antibody in healthy seropositive adults. Volunteers given low-dose SynGEM (140 µg F, 2mg BLP) required a boost at day 28 to achieve plateau responses with a maximum fold-change of 2.4, whereas high-dose recipients (350 µg F, 5mg BLP) achieved plateau responses with a fold-change of 1.5 after first vaccination that remained elevated up to 180 days post-vaccination irrespective of further boosting. Palivizumab-like antibodies were consistently induced, but F protein site Ø-specific antibodies were not detected and virus-specific nasal IgA responses were heterogeneous, with strongest responses in individuals with lower pre-existing antibody levels. CONCLUSIONS:SynGEM is thus the first non-replicating intranasal RSV subunit vaccine to induce persistent antibody responses in human volunteers. Clinical trial registration available at www.clinicaltrials.gov, ID NCT02958540

Real-time amplification of HLA-DQA1 for counting residual white blood cells in filtered platelet concentrates

BACKGROUND A real-time polymerase chain reaction (PCR) assay based on amplification of a conserved region of the HLA-DQA1 locus was developed and validated to assess its suitability in quantitating low levels of white blood cells (WBCs) in filtered platelet (PLT) concentrates (PCs). STUDY DESIGN AND METHODS: To determine the detection limit, serial dilutions of nonfiltered PCs with known quantities of WBCs were prepared. The analytical sensitivity and accuracy of the assay was tested with WBC concentrations ranging from 300 to 0.03 per microL with real-time PCR and flow cytometry. In addition, 126 random PCs were investigated to assess the capacity of the PCR method to quantify residual WBCs in clinical specimens. RESULTS: A sensitivity of 0.2 WBC equivalent per micro L (1.5 x 10(4) WBC equivalents/unit) was achieved. The assay was shown to be accurate and the HLA-DQA1 gene was reproducibly and consistently amplified in all tested samples (coefficient of variance of < 5%). Overall, the results of the PCR assay correlated well with those of the flow cytometry. The PCR assay detected a concentration of 3 WBCs per micro L (approximately 1 x 10(6) WBCs/unit) with 100 percent accuracy. CONCLUSION: Real-time PCR is rapid, sensitive, accurate, and reproducible. Hence this approach may prove suitable in routine monitoring of residual WBCs in PC