11 research outputs found
DNA Encoding an HIV-1 Gag/Human Lysosome-Associated Membrane Protein-1 Chimera Elicits a Broad Cellular and Humoral Immune Response in Rhesus Macaques
Previous studies of HIV-1 p55Gag immunization of mice have demonstrated the usefulness of targeting antigens to the cellular compartment containing the major histocompatibility complex type II (MHC II) complex molecules by use of a DNA antigen formulation encoding Gag as a chimera with the mouse lysosome-associated membrane protein (mLAMP/gag). In the present study, we have analyzed the magnitude and breadth of Gag-specific T-lymphocyte and antibody responses elicited in Rhesus macaques after immunization with DNA encoding a human LAMP/gag (hLAMP/gag) chimera. ELISPOT analyses indicated that the average Gag-specific IFN-γ response elicited by the hLAMP/gag chimera was detectable after only two or three naked DNA immunizations in all five immunized macaques and reached an average of 1000 spot-forming cells (SFC)/10(6) PBMCs. High IFN-γ ELISPOT responses were detected in CD8(+)-depleted cells, indicating that CD4(+) T-cells play a major role in these responses. The T-cell responses of four of the macaques were also tested by use of ELISPOT to 12 overlapping 15-amino acids (aa) peptide pools containing ten peptides each, encompassing the complete Gag protein sequence. The two Mamu 08 immunized macaques responded to eight and twelve of the pools, the Mamu B01 to six, and the other macaque to five pools indicating that the hLAMP/gag DNA antigen formulation elicits a broad T-cell response against Gag. Additionally, there was a strong HIV-1-specific IgG response. The IgG antibody titers increased after each DNA injection, indicating a strong amnestic B-cell response, and were highly elevated in all the macaques after three immunizations. Moreover, the serum of each macaque recognized 13 of the 49 peptides of a 20-aa peptide library covering the complete Gag amino acid sequence. In addition, HIV-1-specific IgA antibodies were present in the plasma and external secretions, including nasal washes. These data support the findings of increased immunogenicity of genetic vaccines encoded as LAMP chimeras, including the response to DNA vaccines by non-human primates
Protective immunity of single and multi-antigen DNA vaccines against schistosomiasis
We evaluated the usefulness of the combination of three plasmids
encoding tegumental (pECL and pSM14) and muscular (pIRV5) antigens of
the Schistosoma mansoni on improving the protective immunity
over the use of a single antigen as DNA vaccines. Female BALB/c mice
were inoculated twice with 25 µg DNA plasmid within two weeks
interval. The challenge was performed with 80 cercarias of a regional
isolate of S. mansoni (SLM) one week after the last immunization. Six
weeks after challenge, all mice were perfused for worm load
determination. The following groups were analyzed: saline; empty
vector; monovalent formulations of pECL; pSM14 and pIRV5 and also
double combinations of pECL/pIRV5 and pIRV5/pSM14 and a triple
combination of pECL/pIRV5/pSM14. The protection was expressed as a
percentage of worm loads in each group compared with the saline group.
The results obtained were 41% (p < 0.05); 52% (p < 0.05); 51% (p
< 0.05); 48% (p < 0.05); 55% (p < 0.05); 45% (p < 0.05);
65% (p < 0.05) for each group respectively
Astrocytic expression of transgene in the rat brain mediated by baculovirus vectors containing an astrocyte-specific promoter
10.1038/sj.gt.3302771Gene Therapy13201447-1456GETH