Characterization of human CD4(+) T-cell clones recognizing conserved and variable epitopes of the Lassa virus nucleoprotein

T cells must play the major role in controlling acute human Lassa virus inf ection, because patients recover from acute Lassa fever in the absence of a measurable neutralizing antibody response. T cells alone seem to protect a nimals from a lethal Lassa virus challenge, because after experimental vacc ination no neutralizing antibodies are detectable. In order to study human T-cell reactivity to single Lassa virus proteins, the nucleoprotein (NP) of Lassa virus, strain Josiah, was cloned, expressed in Escherichia coil, and affinity purified, Peripheral blood mononuclear cells (PBMC) obtained from 8 of 13 healthy, Lassa virus antibody-positive individuals living in the R epublic of Guinea, western Africa, were found to proliferate in response to the recombinant protein (proliferation index greater than or equal to 10). PBMC obtained from one individual with a particularly high proliferative r esponse were used to generate 50 NP-specific T-cell clones (TCC), For six o f these the epitopes were mapped with overlapping synthetic peptides derive d from the sequence of the NP. These CD4(+) TCC displayed high specific pro liferation and produced mainly gamma interferon upon stimulation with NP, B ecause variation of up to 15% in the amino acid sequences of the structural proteins of naturally occurring Lassa virus variants has been observed, th e reactivity of the TCC with peptides derived from the homologous epitopes of the Nigeria strain of Lassa virus and of the eastern Africa arenavirus M opeia was tested. With the Nigeria strain of Lassa virus the levels of homo logy were 100% for two of these epitopes and 85% for three of them, whereas homology with the respective Mopeia epitopes ranged from 92 to 69%, Reacti vity of the TCC with peptides derived from the variable epitopes of the Nig eria strain and of Mopeia was reduced or completely abolished. This report shows for the first time that seropositive individuals from areas of endemi city have very strong memory CD4(+) T-cell responses against the NP of Lass a virus, which are partly strain specific and partly cross-reactive with ot her Lassa virus strains. Our findings may have important implications for t he strategy of designing recombinant vaccines against this mainly T-cell co ntrolled human arenavirus infection.