Carbohydrate fatty acid monosulphate esters are safe and effective adjuvants for humoral responses.

Carbohydrate fatty acid sulphate esters (CFASEs) formulated in a squalane-in-water emulsion are effective adjuvants for humoral responses to a wide range of antigens in various animal species but rise in body temperature and local reactions albeit mild or minimal hampers application in humans. In rabbits, body temperature increased 1°C one day after intramuscular (IM) injection, which returned to normal during the next day. The effect increased with increasing dose of CFASE but not with the number of injections (up to 5). Antigen enhanced the rise in body temperature after booster immunization (P<0.01) but not after priming. Synthetic CFASEs are mixtures of derivatives containing no sulphate, one or multiple sulphate groups and the monosulphate derivatives (CMS) were isolated, incorporated in a squalane in-water emulsion and investigated. In contrast to CFASE, CMS adjuvant did not generate rise in body temperature or local reactions in rabbits immunized with a purified, recombinant malaria chimeric antigen R0.10C. In comparison to alum, CMS adjuvant revealed approximately 30-fold higher antibody titres after the first and >100-fold after the second immunization. In ferrets immunized with 7.5µg of inactivated influenza virus A/H7N9, CMS adjuvant gave 100-fold increase in HAI antibody titres after the first and 25-fold after the second immunisation, which were 10-20-fold higher than with the MF59-like AddaVax adjuvant. In both models, a single immunisation with CMS adjuvant revealed similar or higher titres than two immunisations with either benchmark, without detectable systemic and local adverse effects. Despite striking chemical similarities with monophospholipid A (MPL), CMS adjuvant did not activate human TLR4 expressed on HEK cells. We concluded that the synthetic CMS adjuvant is a promising candidate for poor immunogens and single-shot vaccines and that rise in body temperature, local reactions or activation of TLR4 is not a pre-requisite for high adjuvanticity.

A novel guinea pig model of Chlamydia trachomatis genital tract infection.

Genital Chlamydia trachomatis infections often result in pelvic inflammatory disease and sequelae including infertility and ectopic pregnancies. In addition to the already established murine models, the development of other animal models is necessary to study the safety and efficacy of prototype vaccine candidates. The intravaginal infection of guinea pigs with C. trachomatis has been tested in three independent studies. The first two studies investigated the effect of hormonal treatment of the animals prior to infection with serovars D and E. The results showed that estradiol treatment was required for sustained infection. The third study conducted an immunization-challenge experiment to explore the feasibility of measuring protection in this guinea pig model. C. trachomatis bacteria were sampled using vaginal swabs and measured by qPCR. Using immunohistochemistry the bacteria were detected in the oviducts 19 days post-infection, indicating that the estradiol treatment resulted in ascending infection. Furthermore, immunization of guinea pigs with live EB formulated with ISCOM matrix led to reduction of cervico-vaginal shedding and diminished the severity of pathology. In this study we have developed a new guinea pig model of C. trachomatis female genital tract infection for the purpose of evaluating potential vaccine candidates.