Efficient recognition by rat T cell clones of an epitope of mycobacterial hsp 65 inserted in <i>Escherichia coli</i> outer membrane protein PhoE

Hogervorst, E. J. M.; Agterberg, Marja; Wagenaar, J.A.; Adriaanse, Henriëtte; Boog, C. J. P.; Zee, Ruurd van der; Embden, J. D. A. Van; Eden, Willem van; Tommassen, Jan

doi:10.1002/eji.1830201234

Cited by 22 publications

(9 citation statements)

References 31 publications

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“…We have previously shown in the rat system that purified chimeric PhoE proteins containing class 11-restricted T cell epitopes are, both in vitro and in vivo, presented to CD4+ T cells [24,25].We have now demonstrated that aTh epitope expressed as chimeric PhoE protein in E. coli and attenuated S. typhimurium is efficiently processed and presented to human CD4+ T cells by infected human macrophages.…”

Section: Inhibition Of Presentation Of the Mvf T H Epitope By Cytochamentioning

confidence: 88%

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Intracellular processing and presentation of T cell epitopes, expressed by recombinant Escherichia coli and Salmonella typhimurium, to human T cells

et al. 1995

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Vaccines based on recombinant attenuated bacteria represent a potentially safe and effective immunization strategy. A carrier system was developed to analyze in vitro whether foreign T cell epitopes, inserted in the outer membrane protein PhoE of Escherichia coli and expressed by recombinant bacteria, are efficiently processed and presented via human leukocyte antigen (HLA) class I and II molecules by bacterial infected human macrophages. A well-defined HLA-B27-restricted cytotoxic T cell (CTL) epitope and an HLA-DR53 restricted T helper (Th) epitope of the fusion protein of measles virus were genetically inserted in a surface-exposed region of PhoE, and the chimeric proteins were expressed in E. coli and Salmonella typhimurium. Macrophages infected with both recombinant bacteria presented the Th epitope to the specific CD4+ T cell clone, but failed to present the CTL epitope to the specific CD8+ T cell clone. Presentation of the Th epitope by the infected macrophages was inhibited by cytochalasin D, indicating that phagocytic processing of intact bacteria within infected macrophages was essential for antigen presentation via HLA class II. Presentation of the Th epitope to the CD4+ T cell clone by infected macrophages was blocked by brefeldin A and cycloheximide, indicating the requirement of nascent HLA class II molecules for presentation. The efficiency of macrophages to process and present the inserted Th epitope was similar for both the recombinant E. coli and S. typhimurium strains.

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Section: Inhibition Of Presentation Of the Mvf T H Epitope By Cytochamentioning

confidence: 88%

“…Chimeric proteins consisting of the E. coli PhoE protein with insertions of the Th or CTL epitope of MVF were genetically constructed by the strategy described previously [24]. Restriction enzymes and other DNA modifying enzymes were purchased from Pharmacia (Uppsala, Sweden).…”

Section: Construction and Characterization Of The Chimeric Phoe Proteinsmentioning

confidence: 99%

Intracellular processing and presentation of T cell epitopes, expressed by recombinant Escherichia coli and Salmonella typhimurium, to human T cells

et al. 1995

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“…Using the bacterial PhoE pore-forming protein, Hogervorst et al [28] have previously prepared a chimeric protein, expressing a Tcell epitope of the hsp65 of Mycobacterium tuberculosis. Recognition of the inserted T cell epitope was independent of antigen processing events and was found after stimulation of specific T cells in the presence of fixed APC.…”

Section: Discussionmentioning

confidence: 99%

Induction of T cell responses by chimeric bacterial proteins expressing several copies of a viral T cell epitope

et al. 1993

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A viral T cell epitope was genetically inserted within the periplasmic MalE protein of Escherichia coli in two different permissive insertion sites and resulting hybrid proteins were used to study the in vitro and in vivo immunogenicity of the foreign T cell epitope. Purified hybrid MalE proteins containing the T cell epitope 120-132 (PreS:T) from PreS2 region of hepatitis B virus HBsAg inserted alone or with its adjacent B cell epitope (132-145) were able to induce strong peptide-specific T cell responses in mice. In vitro stimulation of primed lymph node cells or specific T cell hybridomas by the hybrid proteins required processing of the inserted T cell epitope and was inhibited by antigen-presenting cells fixation. The inserted T cell epitope was presented in vitro, in association with appropriate major histocompatibility complex molecules, as efficiently as free synthetic peptide. The in vitro immunogenicity of MalE hybrid proteins was increased by inserting four tandemly repeated copies of PreS:T, either at site 133 or 303. These results were confirmed in vivo by comparing the proliferative responses of lymph node cells from DBA/1 mice primed with MalE hybrid proteins containing one or four copies of PreS:T. Thus, the use of MalE hybrid proteins expressing multiple copies of a given foreign T cell epitope allows the induction of peptide-specific T cell response with a lower dose of priming antigen.

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“…The latter has already been used to insert a well-characterized epitope of the 60-kDa heat shock protein of M. tuberculosis into the cell surface-exposed domain. Semipurified hybrid protein stimulated specific T cell clones [66].…”

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confidence: 99%

Vaccination strategies against intracellular microbes

Hess

Kaufmann

1993

FEMS Immunology &Amp; Medical Microbiology

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Efficient recognition by rat T cell clones of an epitope of mycobacterial hsp 65 inserted in Escherichia coli outer membrane protein PhoE

Cited by 22 publications

References 31 publications

Intracellular processing and presentation of T cell epitopes, expressed by recombinant Escherichia coli and Salmonella typhimurium, to human T cells

Intracellular processing and presentation of T cell epitopes, expressed by recombinant Escherichia coli and Salmonella typhimurium, to human T cells

Induction of T cell responses by chimeric bacterial proteins expressing several copies of a viral T cell epitope

Vaccination strategies against intracellular microbes

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