Characterization of V3 loop-Pseudomonas exotoxin chimeras. Candidate vaccines for human immunodeficiency virus-1
To develop a candidate vaccine for human immunodeficiency virus, type 1 (HIV-1), chimeric proteins were constructed by inserting sequences derived from the V3 loop of gp120 into a nontoxic form of Pseudomonas exotoxin (PE). Inserts of 14 or 26 amino acids, constrained by a disulfide bond, were intro...
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| Veröffentlicht in: | The Journal of biological chemistry 1998-04, Vol.273 (16), p.9951-9958 |
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| container_title | The Journal of biological chemistry |
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| creator | FitzGerald, D J Fryling, C M McKee, M L Vennari, J C Wrin, T Cromwell, M E Daugherty, A L Mrsny, R J |
| description | To develop a candidate vaccine for human immunodeficiency virus, type 1 (HIV-1), chimeric proteins were constructed by inserting sequences derived from the V3 loop of gp120 into a nontoxic form of Pseudomonas exotoxin (PE). Inserts of 14 or 26 amino acids, constrained by a disulfide bond, were introduced between domains II and III of PE. V3 loop-toxin proteins expressed in Escherichia coli and corresponding to either MN (subtype B) or Thai (subtype E) strains, were recognized by strain-specific monoclonal anti-gp120 antibodies. When loop sequences were introduced into an enzymatically active form of the toxin, there was no loss of toxin-mediated cell killing, suggesting that these sequences were co-transported to the cytosol. Sera from rabbits injected with nontoxic PE-V3 loop chimeras were reactive for strain-specific gp120s in Western blots, immunocapture assays, enzyme-linked immunosorbent assays, and neutralized HIV-1 infectivity. Since toxin vectors were designed to receive oligonucleotide duplexes encoding any V3 loop sequence, this approach should allow for the production of V3 loop-toxin chimeras corresponding to multiple HIV isolates. |
| doi_str_mv | 10.1074/jbc.273.16.9951 |
| format | Article |
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V3 loop-toxin proteins expressed in Escherichia coli and corresponding to either MN (subtype B) or Thai (subtype E) strains, were recognized by strain-specific monoclonal anti-gp120 antibodies. When loop sequences were introduced into an enzymatically active form of the toxin, there was no loss of toxin-mediated cell killing, suggesting that these sequences were co-transported to the cytosol. Sera from rabbits injected with nontoxic PE-V3 loop chimeras were reactive for strain-specific gp120s in Western blots, immunocapture assays, enzyme-linked immunosorbent assays, and neutralized HIV-1 infectivity. Since toxin vectors were designed to receive oligonucleotide duplexes encoding any V3 loop sequence, this approach should allow for the production of V3 loop-toxin chimeras corresponding to multiple HIV isolates.</description><identifier>ISSN: 0021-9258</identifier><identifier>DOI: 10.1074/jbc.273.16.9951</identifier><identifier>PMID: 9545339</identifier><language>eng</language><publisher>United States</publisher><subject>Acquired Immunodeficiency Syndrome - blood ; Acquired Immunodeficiency Syndrome - immunology ; ADP Ribose Transferases ; AIDS Vaccines ; Amino Acid Sequence ; Animals ; Bacterial Toxins ; Carcinoma, Squamous Cell ; Cell Survival - drug effects ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; Exotoxins - chemistry ; Exotoxins - immunology ; Exotoxins - toxicity ; HIV Antibodies - blood ; HIV Antibodies - immunology ; HIV Envelope Protein gp120 - chemistry ; HIV Envelope Protein gp120 - immunology ; HIV Envelope Protein gp120 - toxicity ; HIV-1 - drug effects ; HIV-1 - isolation & purification ; Humans ; Molecular Sequence Data ; Neutralization Tests ; Protein Conformation ; Pseudomonas aeruginosa ; Pseudomonas aeruginosa Exotoxin A ; Rabbits ; Recombinant Fusion Proteins - chemistry ; Recombinant Fusion Proteins - immunology ; Recombinant Fusion Proteins - toxicity ; Tumor Cells, Cultured ; Vaccines, Synthetic ; Virulence Factors</subject><ispartof>The Journal of biological chemistry, 1998-04, Vol.273 (16), p.9951-9958</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9545339$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>FitzGerald, D J</creatorcontrib><creatorcontrib>Fryling, C M</creatorcontrib><creatorcontrib>McKee, M L</creatorcontrib><creatorcontrib>Vennari, J C</creatorcontrib><creatorcontrib>Wrin, T</creatorcontrib><creatorcontrib>Cromwell, M E</creatorcontrib><creatorcontrib>Daugherty, A L</creatorcontrib><creatorcontrib>Mrsny, R J</creatorcontrib><title>Characterization of V3 loop-Pseudomonas exotoxin chimeras. Candidate vaccines for human immunodeficiency virus-1</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>To develop a candidate vaccine for human immunodeficiency virus, type 1 (HIV-1), chimeric proteins were constructed by inserting sequences derived from the V3 loop of gp120 into a nontoxic form of Pseudomonas exotoxin (PE). Inserts of 14 or 26 amino acids, constrained by a disulfide bond, were introduced between domains II and III of PE. V3 loop-toxin proteins expressed in Escherichia coli and corresponding to either MN (subtype B) or Thai (subtype E) strains, were recognized by strain-specific monoclonal anti-gp120 antibodies. When loop sequences were introduced into an enzymatically active form of the toxin, there was no loss of toxin-mediated cell killing, suggesting that these sequences were co-transported to the cytosol. Sera from rabbits injected with nontoxic PE-V3 loop chimeras were reactive for strain-specific gp120s in Western blots, immunocapture assays, enzyme-linked immunosorbent assays, and neutralized HIV-1 infectivity. Since toxin vectors were designed to receive oligonucleotide duplexes encoding any V3 loop sequence, this approach should allow for the production of V3 loop-toxin chimeras corresponding to multiple HIV isolates.</description><subject>Acquired Immunodeficiency Syndrome - blood</subject><subject>Acquired Immunodeficiency Syndrome - immunology</subject><subject>ADP Ribose Transferases</subject><subject>AIDS Vaccines</subject><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Bacterial Toxins</subject><subject>Carcinoma, Squamous Cell</subject><subject>Cell Survival - drug effects</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Escherichia coli</subject><subject>Exotoxins - chemistry</subject><subject>Exotoxins - immunology</subject><subject>Exotoxins - toxicity</subject><subject>HIV Antibodies - blood</subject><subject>HIV Antibodies - immunology</subject><subject>HIV Envelope Protein gp120 - chemistry</subject><subject>HIV Envelope Protein gp120 - immunology</subject><subject>HIV Envelope Protein gp120 - toxicity</subject><subject>HIV-1 - drug effects</subject><subject>HIV-1 - isolation & purification</subject><subject>Humans</subject><subject>Molecular Sequence Data</subject><subject>Neutralization Tests</subject><subject>Protein Conformation</subject><subject>Pseudomonas aeruginosa</subject><subject>Pseudomonas aeruginosa Exotoxin A</subject><subject>Rabbits</subject><subject>Recombinant Fusion Proteins - chemistry</subject><subject>Recombinant Fusion Proteins - immunology</subject><subject>Recombinant Fusion Proteins - toxicity</subject><subject>Tumor Cells, Cultured</subject><subject>Vaccines, Synthetic</subject><subject>Virulence Factors</subject><issn>0021-9258</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1998</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNotkDtPwzAAhD2ASinMTEie2BLiR5x4RBEvqRIMwBr5qbqK7WAnVeHXE4necsunT7oD4AZVJaoaer-XqsQNKRErOa_RGVhXFUYFx3V7AS5z3ldLKEcrsOI1rQnhazB2O5GEmkxyv2JyMcBo4ReBQ4xj8Z7NrKOPQWRojnGKRxeg2jlvksgl7ETQTovJwINQygWToY0J7mYvAnTezyFqY51yJqgfeHBpzgW6AudWDNlcn3oDPp8eP7qXYvv2_No9bIsRk2YqrKzbpqGGY0Z1i4VdBkqNa2uVwRbblmGOdSNbxhRhkmvJULvQHDWtxNaSDbj7944pfs8mT713WZlhEMHEOfeIEcopowt4ewJn6Y3ux-S8SD_96SLyByo6aM0</recordid><startdate>19980417</startdate><enddate>19980417</enddate><creator>FitzGerald, D J</creator><creator>Fryling, C M</creator><creator>McKee, M L</creator><creator>Vennari, J C</creator><creator>Wrin, T</creator><creator>Cromwell, M E</creator><creator>Daugherty, A L</creator><creator>Mrsny, R J</creator><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7U9</scope><scope>H94</scope></search><sort><creationdate>19980417</creationdate><title>Characterization of V3 loop-Pseudomonas exotoxin chimeras. Candidate vaccines for human immunodeficiency virus-1</title><author>FitzGerald, D J ; Fryling, C M ; McKee, M L ; Vennari, J C ; Wrin, T ; Cromwell, M E ; Daugherty, A L ; Mrsny, R J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p237t-fb58774e9264d82af074bd25ffce2f2f86292d7b866c36b9db618e929178b2ff3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1998</creationdate><topic>Acquired Immunodeficiency Syndrome - blood</topic><topic>Acquired Immunodeficiency Syndrome - immunology</topic><topic>ADP Ribose Transferases</topic><topic>AIDS Vaccines</topic><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Bacterial Toxins</topic><topic>Carcinoma, Squamous Cell</topic><topic>Cell Survival - drug effects</topic><topic>Enzyme-Linked Immunosorbent Assay</topic><topic>Escherichia coli</topic><topic>Exotoxins - chemistry</topic><topic>Exotoxins - immunology</topic><topic>Exotoxins - toxicity</topic><topic>HIV Antibodies - blood</topic><topic>HIV Antibodies - immunology</topic><topic>HIV Envelope Protein gp120 - chemistry</topic><topic>HIV Envelope Protein gp120 - immunology</topic><topic>HIV Envelope Protein gp120 - toxicity</topic><topic>HIV-1 - drug effects</topic><topic>HIV-1 - isolation & purification</topic><topic>Humans</topic><topic>Molecular Sequence Data</topic><topic>Neutralization Tests</topic><topic>Protein Conformation</topic><topic>Pseudomonas aeruginosa</topic><topic>Pseudomonas aeruginosa Exotoxin A</topic><topic>Rabbits</topic><topic>Recombinant Fusion Proteins - chemistry</topic><topic>Recombinant Fusion Proteins - immunology</topic><topic>Recombinant Fusion Proteins - toxicity</topic><topic>Tumor Cells, Cultured</topic><topic>Vaccines, Synthetic</topic><topic>Virulence Factors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>FitzGerald, D J</creatorcontrib><creatorcontrib>Fryling, C M</creatorcontrib><creatorcontrib>McKee, M L</creatorcontrib><creatorcontrib>Vennari, J C</creatorcontrib><creatorcontrib>Wrin, T</creatorcontrib><creatorcontrib>Cromwell, M E</creatorcontrib><creatorcontrib>Daugherty, A L</creatorcontrib><creatorcontrib>Mrsny, R J</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>FitzGerald, D J</au><au>Fryling, C M</au><au>McKee, M L</au><au>Vennari, J C</au><au>Wrin, T</au><au>Cromwell, M E</au><au>Daugherty, A L</au><au>Mrsny, R J</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Characterization of V3 loop-Pseudomonas exotoxin chimeras. 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When loop sequences were introduced into an enzymatically active form of the toxin, there was no loss of toxin-mediated cell killing, suggesting that these sequences were co-transported to the cytosol. Sera from rabbits injected with nontoxic PE-V3 loop chimeras were reactive for strain-specific gp120s in Western blots, immunocapture assays, enzyme-linked immunosorbent assays, and neutralized HIV-1 infectivity. Since toxin vectors were designed to receive oligonucleotide duplexes encoding any V3 loop sequence, this approach should allow for the production of V3 loop-toxin chimeras corresponding to multiple HIV isolates.</abstract><cop>United States</cop><pmid>9545339</pmid><doi>10.1074/jbc.273.16.9951</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | The Journal of biological chemistry, 1998-04, Vol.273 (16), p.9951-9958 |
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| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | Acquired Immunodeficiency Syndrome - blood Acquired Immunodeficiency Syndrome - immunology ADP Ribose Transferases AIDS Vaccines Amino Acid Sequence Animals Bacterial Toxins Carcinoma, Squamous Cell Cell Survival - drug effects Enzyme-Linked Immunosorbent Assay Escherichia coli Exotoxins - chemistry Exotoxins - immunology Exotoxins - toxicity HIV Antibodies - blood HIV Antibodies - immunology HIV Envelope Protein gp120 - chemistry HIV Envelope Protein gp120 - immunology HIV Envelope Protein gp120 - toxicity HIV-1 - drug effects HIV-1 - isolation & purification Humans Molecular Sequence Data Neutralization Tests Protein Conformation Pseudomonas aeruginosa Pseudomonas aeruginosa Exotoxin A Rabbits Recombinant Fusion Proteins - chemistry Recombinant Fusion Proteins - immunology Recombinant Fusion Proteins - toxicity Tumor Cells, Cultured Vaccines, Synthetic Virulence Factors |
| title | Characterization of V3 loop-Pseudomonas exotoxin chimeras. Candidate vaccines for human immunodeficiency virus-1 |
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