DNA and the immune system

DNA and the immune system. member of the subfamily and the causative agent Curcumol of infectious bovine rhinotracheitis and infectious pustular vulvovaginitis (79). BHV-1 has also been shown to cause conjunctival infections, abortions, meningo-encephalitic diseases, and infectious balanoposthitis (30, 84). BHV-1 may be the primary viral agent involved in the development of secondary opportunistic bacterial infections leading to shipping fever in cattle (4, 70, 86). Of the 13 proteins associated with the viral lipid envelope, glycoproteins B (gB), C (gC), and D (gD) are consistently recognized by convalescent-phase sera from BHV-1-infected animals (15, 78). Immunization of cattle with each of these individual glycoproteins, formulated with a conventional adjuvant, leads to protective immune responses that include neutralizing serum antibodies and cell-mediated immunity (CMI) (3, 35, 71, 79). Immunization of cattle with gD typically results in the humoral responses of the greatest magnitude (3, 81). Also, gD Curcumol has been demonstrated to efficiently induce CMI and humoral immune responses in C57BL/6 (promoter (69). The appropriate orientation of the ligation product created a novel start codon contributed by pAA505 followed by Asp and Pro prior to in-frame commencement of the gD amino acid sequence Tyr Val Asp Pro immediately downstream from the signal peptide sequence. Clone pAACgD was digested with DNA polymerase I, and blunt end ligated to a 0.05) lower than the 2- and 4-week titers in mice immunized with pSLRSV.AgD and significantly ( 0.05) lower than the 2-, 4-, and 6-week titers of mice immunized with pSLRSV.SgD (Fig. ?(Fig.4).4). Physique Rabbit polyclonal to CD24 (Biotin) ?Physique44 also demonstrates that all Curcumol plasmid-immunized Curcumol groups induce antibodies of similar magnitude to an authentic gD subunit protein formulated in the adjuvant VSA3. Open in a separate windows FIG. 3 Kinetics of serum anti-BHV-1 gD antibodies in C3H/HeN mice immunized with plasmids encoding cell-associated or secreted forms of BHV-1 gD. Each DNA-based vaccine group was comprised of five mice. Each mouse received 100 g Curcumol (2 g/l in normal saline) of plasmid DNA i.m. in the left quadriceps muscle mass on days 0 and 14. Serum ELISA titers for individual mice were determined by using the extrapolation function (Microsoft Excel) based on endpoint dilutions and with preimmune serum means (plus 3 standard deviations) as cutoffs. Endpoint titers are expressed as 1/log10. Serum antibody levels were decided for individual mice at each time point, except at 2 weeks, when blood samples for each group were pooled. Data are expressed as the geometric mean of four (pSLRSV.AgD) or two (pSLRSV.CgD) seropositive mice. Error bars show the standard error of the means. Only 23-week titers from mice that were seropositive at week 14 were determined. Open in a separate windows FIG. 4 Serum antibody levels after immunization of C3H/HeN mice with plasmids encoding cell-associated (AgD or CgD) or secreted (SgD) antigens. Experimental groups consisted of 10 mice and included a null plasmid control group and a subunit gD vaccine group. Mice 6 to 7 weeks of age were immunized in each quadriceps muscle mass with 50 g of DNA (total of 100 g) or subcutaneously with 400 ng of affinity-purified rtgD in 100 l of VSA3 plus HBSS. All mice were given a booster injection at 2 weeks after initial immunization with the same dose. Serum ELISA titers were determined as described for Fig. ?Fig.3.3. Plasmid construct or subunit gD designations are indicated in the top center of each.