Enhancement of humoral immune responses to a human cytomegalovirus DNA vaccine: Adjuvant effects of aluminum phosphate and CpG oligodeoxynucleotides

Abstract

A human cytomegalovirus (HCMV) glycoprotein B (gpUL55) DNA vaccine has been evaluated in BALB/c mice. Intramuscular immunization of these mice with pRc/CMV2-gB resulted in the generation of high levels of gpUL55-specific antibody (geometric mean titer [GMT] 1:8900) and neutralizing antibody (GMT 1:74) after 2 booster doses given 5 and 10 weeks after primary inoculation. Emulsifying the construct with the aluminum phosphate gel adjuvant Adju-Phos before immunization enhanced gpUL55-specific antibody responses (GMT 1:17800, P = 0.04). Co-immunization with CpG oligodeoxynucleotides was shown to enhance levels of neutralizing antibodies generated by immunization of mice with a pRc/CMV2-gB/Adju-Phos emulsion (P = 0.04). The results provide a rationale for evaluating combinations of other HCMV proteins for incorporation into a multi-target DNA vaccine, and for the optimization of adjuvant usage, to elicit enhanced levels of neutralizing antibodies. 2003.

Distribution of UL144, US28 and UL55 genotypes in Polish newborns with congenital cytomegalovirus infections

“…Briefly, following activation for 10 min at 95°C, 60 cycles of 15 s at 95°C and 1 min at 60°C were carried out in a 7900HT Fast Real-Time PCR System (Applied Biosystems, USA). Triplicate standards of plasmid DNA containing the entire UL55 open reading frame and consisting of 1, 5, 10, 100, 1,000, 10,000 and 100,000 GE (genome equivalents) were included in each real-time analysis [41]. The number of target copies in the reaction was deduced from the threshold cycle C T ).…”

Immunomodulatory effect of plasmids co-expressing cytokines in classical swine fever virus subunit gp55/E2-DNA vaccination

“…Vaccination with E2-DNA allows the induction of a long term immune response and as mentioned above the serological differentiation between vaccinated and infected animals [44]. Nevertheless the immunogenicity of the DNAconstructs is sometimes low and the immunostimulatory capacity might be improved by other components like cytokine genes or CpG-motifs [4,20,40].…”

Kinetics of Cytomegalovirus Load Decrease in Solid‐Organ Transplant Recipients after Preemptive Therapy with Valganciclovir

“…In addition each TaqMan PCR run contained a dilution series of cloned HCMV gB DNA in triplicate of 1-10 4 genomes [13]. PCR cycling conditions were 2 min at 50ЊC, 10 min at 95ЊC, and 60 cycles of 15 s at 95ЊC and 15 s at 60ЊC.…”