Important note: Information in this article was accurate in 1990. The state of the art may have changed since the publication date.
HIV RECOMBINANT PROTEINS: IMMUNOLOGICAL ANALYSIS AND APPLICATIONS AS REAGENTS FOR SEROLOGICAL TESTS
HIV Detection by Genetic Engineering Methods. Luciw PA and Steimer KS, eds. New York, Marcel Dekker, p. 59-76, 1989.. Unique Identifier : AIDSLINE ICDB/90660255 Petteway SR Jr; Reed DL; Tribe DE; Kenealy WR; E.I. du Pont de Nemours and Co., Wilmington, DE
Abstract:
Current blood screening tests for HIV make use of viral lysates in an ELISA format to detect past exposure to viral antigens. Control of the viral antigens in these preparations is difficult, making interpretation problematic. To obtain sufficient quantities of pure antigen, recombinant HIV gag (GAG55) and env (ENV9) proteins were produced in Escherichia coli. Immunologic analyses of these biosynthetic viral proteins and their use as reagents in serologic testing are described. GAG55 represents the p55 core precursor protein. ENV9 is a recombinant protein that represents the carboxy terminus of gp120 fused to the amino acid terminus of gp41. Both GAG55 and ENV9 proteins reacted with monoclonal and polyclonal antibodies specific for HIV proteins using Western blot and ELISA formats. In immunoblot competition experiments, GAG55 was able to compete out the reactivity in HIV-positive human serum to p55, p24, p17, and a 9 kD protein. Monoclonal antibodies to synthetic peptides and viral proteins that reacted with viral antigens also were reactive with ENV9. Most monoclonal antibodies specific for gag or env proteins tested so far react with the respective recombinant proteins. GAG55- and ENV9-specific antisera reacted with the expected proteins on viral immunoblots. The antisera also were reactive with live HIV-infected cells. Anti-ENV9 showed a surface fluorescence pattern and was as effective as antisera to native gp120 in fluorescence-activated cell-sorting analysis and in precipitating purified gp120 in radioimmunoprecipitation assays. Recombinant proteins were used in an ELISA format to investigate the dynamics of the host immune response to HIV and to analyze their potential as test reagents for the detection of HIV antibodies in infected individuals. It appears that, while most infected persons maintain antibodies reactive with envelope antigens, as many as 50% demonstrate little or no reactivity to gag. These studies suggest that the recombinant proteins produced in E coli share important antigenic and immunogenic structures with natural viral proteins produced during HIV infection. Furthermore, they provide a basis for serologic tests with increased sensitivity and specificity compared to tests using viral lysates. (40 Refs)
Keywords: AIDS Serodiagnosis/*METHODS Base Sequence *Cloning, Molecular Gene Expression Regulation, Viral Gene Products, gag/GENETICS Human HIV/*GENETICS/IMMUNOLOGY HIV Antigens/GENETICS/IMMUNOLOGY HIV Envelope Protein gp120/GENETICS HIV Envelope Protein gp41/GENETICS HIV Infections/*DIAGNOSIS/IMMUNOLOGY Protein Precursors/GENETICS MONOGRAPH REVIEW REVIEW, TUTORIAL 900530
M9051020
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Important note: Information in this article was accurate in 1990. The state of the art may have changed since the publication date.
