HIGH-LEVEL EXPRESSION OF THE MEASLES-VIRUS NUCLEOCAPSID PROTEIN BY USING A REPLICATION-DEFICIENT ADENOVIRUS VECTOR - INDUCTION OF AN MHC-1-RESTRICTED CTL RESPONSE AND PROTECTION IN A MURINE MODEL

Replication-deficient adenovirus (Ad) vectors provide an efficient tec hnology for direct DNA delivery to cells both in vitro and in vivo. We have inserted the measles virus nucleoprotein (N) gene under the cont rol of the strong constitutive CMV major IE promoter into an Ad type 5 E1(-) vector to produce the recombinant virus RAd68. Following infect ion of human fibroblasts with RAd68 in vitro, recombinant N protein wa s synthesized as a 60-kDa protein that represented up to 20% total sol uble cell protein. Long filamentous structures were produced in both t he nucleus and the cytoplasm that were similar in appearance to measle s virus nucleocapsids. These ''nucleocapsid-like'' structures were rea dily purified by density gradient centrifugation. Murine immunization with RAd68 elicited (i) a humoral immune response to N, (ii) a major h istocompatibility complex class I-restricted, antigen-specific cytotox ic T cell response, and (iii) protection against challenge with the me asles virus CAM/RB strain in mice. This study demonstrates the capacit y of replication-deficient Ad recombinants both to induce and to chara cterize cell-mediated immune responses. (C) 1995 Academic Press, Inc.