Fine mapping of HIV-1 Nef-epitopes by monoclonal antibodies

Siakkou, H; Jahn, S.; Kienzle, Norbert; Ulrich, Rainer G.; Grötzinger, Carsten; Schneider, Thomas; Kohleisen, Birgit; Pauli, Georg; Spohn, Renate; Jung, Günther; Müller-Lantzsch, N.; Krüger, Detlev H.

doi:10.1007/bf01309790

Cited by 6 publications

(1 citation statement)

References 46 publications

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“…When the Nef B-cell epitopes of the immunocompetent mice were compared, the clearest similarities were seen in the C terminus. This region seems to be the main common immunogenic region of this protein (10,29,36,38).…”

Section: Discussionmentioning

confidence: 96%

Recognition of prominent viral epitopes induced by immunization with human immunodeficiency virus type 1 regulatory genes

Hinkula

Svanholm

Schwartz

et al. 1997

J Virol

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Mice immunized with the regulatory genes nef, rev, and tat from human immunodeficiency virus type 1 developed both humoral and cellular immune responses to the gene products Nef, Rev, and Tat. This study demonstrates that it is feasible to induce immune reactions to all of these regulatory gene products. Humoral responses were seen after DNA boosts, while potent T-cell proliferative responses were noted already after a single immunization. A Th1-directed immune response was demonstrated early after immunization. A 3-to 75-fold-stronger T-cell response was seen in animals receiving DNA epidermally compared to that in animals receiving intramuscular injections. Nef, Rev, and Tat putative Band T-cell epitopes were clearly mapped by using peptides derived from the regulatory proteins and were similar to those which are detected in human immunodeficiency virus infection. Although immunization by the Nef, Rev, and Tat proteins raised high immunoglobulin G titers in serum, the epitope spreading appeared broader after DNA immunization. The combination of all of these regulatory genes together with two genes for structural proteins, the envelope and gag genes, demonstrated that a combined approach is feasible in that reactivities to all antigens persisted or were even augmented. No interference between plasmids was noted.

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Section: Discussionmentioning

confidence: 96%

Recognition of prominent viral epitopes induced by immunization with human immunodeficiency virus type 1 regulatory genes

Hinkula

Svanholm

Schwartz

et al. 1997

J Virol

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Hamster polyomavirus-encoded proteins: Gene cloning, heterologous expression and immunoreactivity

et al. 1996

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The hamster polyomavirus (HaPV) is associated with spontaneously appearing skin epithelioma of the Syrian hamster Z3 strain. Virus particles prepared from the skin epithelioma cause lymphoma and leukemia when injected into newborn hamsters from a distinct Syrian hamster colony (HaP); in contrast to the skin epithelioma the hemopoietic tumors are virus free but accumulate viral DNA. To study the humoral immune response of HaPV-infected Z3 hamsters we produced recombinant HaPV proteins in Escherichia coli as beta-galactosidase-, TrpE- and dihydrofolate reductase-fusion proteins or as non-fused proteins. Recombinant plasmids carried segments of all putative early and late HaPV proteins. The recombinant proteins were detected in stained SDS polyacrylamide gels and in Western blots using monoclonal anti-TrpE and anti-beta-galactosidase antibodies and sera of HaPV-infected hamsters. Sera from HaPV-infected Z3 hamsters and crude lysates of all clones were applied to Western blots to characterize the humoral immune response in the animals. HaPV-specific antibodies were found to be directed against early protein segments translated from the first common exon and from the second unique exon of LT and MT, resp., as well as against the late proteins VP1 and VP2/3. The almost complete VP2 was recognized by all sera whereas VP1 was detected only by a half of the sera. Our data suggest the presence of at least 2 immunodominant regions in VP2, one in the C-terminal VP1 and at least 4 in early proteins.

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Subcloning of HIV-2nef genes inE. coli and immunological reactivity of expressed fusion proteins

et al. 1993

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The nef gene located in the 3' region of the HIV-2 genome encodes an N-terminally myristylated protein of 27-35 kD, likely to be involved in the regulation of viral transcription. The nef genes of HIV-2 isolates GH-1, ROD, ST, BEN, and D194.17 were inserted into E. coli pEX vectors and expression of Nef beta-galactosidase fusion proteins was detected in stained gels. All fusion proteins specifically reacted with a rabbit serum raised against bacterially expressed Nef from HIV-2D194.17. Sera from monkeys inoculated with HIV-2BEN or SIVMAC251 recognized the Nef proteins of only certain HIV-2 isolates. No cross-reactivity of these sera with HIV-1 Nef and of a rabbit anti-HIV-1-Nef serum with the described HIV-2 Nef fusion proteins was observed.

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Fine mapping of HIV-1 Nef-epitopes by monoclonal antibodies

Cited by 6 publications

References 46 publications

Recognition of prominent viral epitopes induced by immunization with human immunodeficiency virus type 1 regulatory genes

Recognition of prominent viral epitopes induced by immunization with human immunodeficiency virus type 1 regulatory genes

Hamster polyomavirus-encoded proteins: Gene cloning, heterologous expression and immunoreactivity

Subcloning of HIV-2nef genes inE. coli and immunological reactivity of expressed fusion proteins

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