Important note: Information in this article was accurate in 1988. The state of the art may have changed since the publication date.
EXPRESSION OF HTLV-III ENVELOPE PROTEINS IN ESCHERICHIA COLI AND THEIR INDUCTION OF NEUTRALIZING ANTIBODIES
Vaccines 87. Modern Approaches to New Vaccines: Prevention of AIDS and Other Viral, Bacterial, and Parasitic Diseases. Chanock RM et al, eds. New York, Cold Spring Harbor Laboratory, p. 256-9, 1987.. Unique Identifier : AIDSLINE ICDB/88647987 Putney SD; Javaherian K; Jackson J; Lynn D; Rusche J; Mueller WT; Matthews T; Bolognesi D; Ghrayeb J; Chanda PK; et al; RepliGen Corp., Cambridge, MA 02139
Abstract:
Several different approaches are being taken toward the development of an acquired immune deficiency syndrome (AIDS) vaccine. The most promising of these appears to be immunization with a form of the outer viral envelope protein, gp120, and recombinant vehicles engineered to express this protein have included Escherichia coli, yeast, mammalian cells, and vaccinia virus. The envelope protein gp120 is heavily glycosylated, and there has been much speculation as to the role of glycosylation in viral replication and cytotoxicity and the necessity of the carbohydrate for eliciting neutralizing antibodies that block viral infection. Two segments of gp120 produced in E coli elicit neutralizing antibodies in goats or rabbits. Because E coli is unable to glycosylate proteins, this means that glycosylation is not required to elicit human T-lymphotropic virus type III (HTLV-III) neutralizing antibodies. A map of gp120 and the location of recombinant proteins produced in E coli are shown. HTLV-III envelope protein fragments from E coli and neutralizing antibodies to these proteins were purified. The proteins were used to immunize either rabbits or goats, and strong immune response to these proteins was observed. Rabbit polyclonal antibodies to PB1 react with native gp120 on Western blots. Similar results were obtained with proteins PE3 and PENV9, and antisera to each of these proteins precipitate gp120 to similar extents in a radioimmunoprecipitation assay. Some of these recombinant gp120 proteins were used to produce monoclonal antibodies. The antibodies were screened for their reactivity to native gp120, and several were obtained that specifically recognize the native envelope protein. Results with two of these antibodies indicate that nonglycosylated E coli-produced envelope proteins elicit antibodies that specifically recognize gp120. Polyclonal antisera were assayed for their ability to neutralize HTLV-III infection in vitro. The goat and rabbit antibodies to PB1 had neutralizing titers comparable to those elicited by native gp120. No neutralization was observed with antibodies to PE3 or PENV9, and this suggests that the major neutralizing epitopes are located in the carboxy-terminal half of gp120. (4 Refs)
Keywords: Animal Antibodies, Viral/*ANALYSIS Antibody Specificity Escherichia coli/GENETICS Goats HIV/GENETICS/*IMMUNOLOGY Neutralization Tests Rabbits Retroviridae Proteins/IMMUNOLOGY Vaccines, Synthetic/*IMMUNOLOGY Viral Envelope Proteins/*IMMUNOLOGY Viral Vaccines/*IMMUNOLOGY MEETING PAPER
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Important note: Information in this article was accurate in 1988. The state of the art may have changed since the publication date.
