Enhanced cell surface expression, immunogenicity and genetic stability resulting from a spontaneous truncation of HIV Env expressed by a recombinant MVA.
During propagation of modified vaccinia virus Ankara (MVA) encoding HIV 89.6 Env, a few viral foci stained very prominently. Virus cloned from such foci replicated to higher titers than the parent and displayed enhanced genetic stability on passage. Sequence analysis showed a single nucleotide deletion in the 89.6 env gene of the mutant that caused a frame shift and truncation of 115 amino acids from the cytoplasmic domain. The truncated Env was more highly expressed on the cell surface, induced higher antibody responses than the full-length Env, reacted with HIV neutralizing monoclonal antibodies and mediated CD4/co-receptor-dependent fusion. Intramuscular (i.m.), intradermal (i.d.) needleless, and intrarectal (i.r.) catheter inoculations gave comparable serum IgG responses. However, intraoral (i.o.) needleless injector route gave the highest IgA in lung washings and i.r. gave the highest IgA and IgG responses in fecal extracts. Induction of CTL responses in the spleens of individual mice as assayed by intracellular cytokine staining was similar with both the full-length and truncated Env constructs. Induction of acute and memory CTL in the spleens of mice immunized with the truncated Env construct by i.d., i.o., and i.r. routes was comparable and higher than by the i.m. route, but only the i.r. route induced CTL in the gut-associated lymphoid tissue. Thus, truncation of Env enhanced genetic stability as well as serum and mucosal antibody responses, suggesting the desirability of a similar modification in MVA-based candidate HIV vaccines.
Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA. lwyatt@niaid.nih.gov
Linda S
LS
Wyatt
Igor M
IM
Belyakov
Patricia L
PL
Earl
Jay A
JA
Berzofsky
Bernard
B
Moss
D016915:AIDS Vaccines / Q000276:immunology; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D002460:Cell Line; D002462:Cell Membrane / Q000821:virology*; D002642:Chick Embryo; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D015967:Gene Expression Regulation, Viral; D006678:HIV / Q000235:genetics* / Q000276:immunology* / Q000502:physiology; D007070:Immunoglobulin A / Q000378:metabolism; D007074:Immunoglobulin G / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D009154:Mutation; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014616:Vaccinia virus / Q000235:genetics*; D054299:env Gene Products, Human Immunodeficiency Virus / Q000235:genetics / Q000378:metabolism*
D016915:AIDS Vaccines; D000914:Antibodies, Viral; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D054299:env Gene Products, Human Immunodeficiency Virus