Objective: To investigate the immune responses elicited by pJME with or without various forms of granulocyte-macrophage colony-stimulating factor (GM-CSF) gene. Methods: The changes of the T lymphocyte subsets and the levels of Th cell intracellular cytokines IFN-γ and IL-4 were evaluated by flow cytometric analysis. The cytotoxic T lymphocyte kill activity was assessed by lactate dehydrogenase activity release test. An 80% plaque reduction neutralization test was performed to titrate the neutralization antibody before and after viral challenge. Results: We demonstrated that simultaneous administration of pJME plus plasmid-ecoded GM-CSF (pGM-CSF) activated Th1 immune responses similar to those found by injecting pGM-CSF i.m. into mice 3 days before pJME vaccination, and enhancement of Th2 immunity predominated when the pGM-CSF was injected 3 days after pJME vaccination. Furthermore, the immunization with DNA vaccine encoding precursor membrane envelope/GM-CSF fusion protein was more effective in generating immune responses than that induced by immunization with pJME alone or in combination with pGM-CSF. Conclusions: These observations support the potential of GM-CSF DNA adjuvant for the Th1/Th2 balance and the enhancement of immune responses by showing that the timing of the administration of pGM-CSF and the application of different forms of GM-CSF gene influence the outcome of the resultant immune responses.

Background: Lemierre’s syndrome is a rare and serious complication of pharyngitis with an estimated annual incidence of 1 in 100,000 people worldwide. It is characterized by septic thrombophlebitis of the internal jugular vein with metastatic infection, usually after oropharyngeal infection. Rare cases of Lemierre’s syndrome have been reported to be caused by odontogenic infection.Case report: A 33-year-old male visited our hospital with symptoms of fever and sore throat for 16 days. The other symptoms included pain in his left neck and shoulder. In addition, metabolic syndrome was diagnosed based on waist circumference, diabetes, and hyperlipidemia. Fusobacterium necrophorum bacteria was detected using the metagenomic next-generation sequencing (mNGS) technique. The enhanced computerized tomography (CT) scan showed thrombosis of the left proximal jugular vein and brachiocephalic vein. Based on these observations, Lemierre’s syndrome was diagnosed. The etiology was that the fillings in the root canal tooth were lost with no blood or pain about 2 weeks before the onset. The patient recovered after treatment with antibiotics and blood purification.Conclusion: Lemierre’s syndrome should be evaluated for patients with fever, sore throat, and neck pain. If the loss of fillings from root canal therapy occurs, especially for those with metabolic syndrome, we should be aware of the possibility of this disease. Furthermore, the mNGS test can be used as a crucial supplementary diagnostic tool for patients with undetermined fever.