Analysis of the primary T-cell response to Sendai virus infection in C57BL/6 mice: CD4+ T-cell recognition is directed predominantly to the hemagglutinin-neuraminidase glycoprotein

Abstract

Sendai virus infection of C57BL/6 mice elicits a strong CD4+ and CD8+ T-cell response in the respiratory tract. To investigate the specificity of the CD4+ T-cell response, a panel of hybridomas was generated from cells recovered from the respiratory tracts of infected mice. Using vaccinia virus recombinants expressing individual Sendai virus proteins, we found that the majority of these hybridomas (34 of 37) were specific for the hemagglutinin-neuraminidase (HN) glycoprotein. The hybridomas were then analyzed for reactivity to a set of overlapping peptides spanning the entire length of the hemagglutinin-neuraminidase glycoprotein. At least five H-2 I-Ab-restricted epitopes were defined in HN. The strong bias toward recognition of class II epitopes derived from a single viral protein contrasts with T-cell recognition of epitopes of several proteins in influenza A virus as found previously by others.

Expansion of CD4 T cells expressing a highly restricted TCR structure specific for a single parasite epitope correlates with high pathology in murine schistosomiasis

“…There are, nevertheless, numerous reports of immunodominance in the literature. Indeed, prominent T cell responses (both CD4 and CD8) have been found to be directed against immunogenic epitopes from nominal antigens [32,33], self antigens that are targets in autoimmune disease [34], viruses [35][36][37][38][39][40][41][42], bacteria [43] and fungi [44]. However, a most relevant example of immunodominance in the area of immunoparasitology is the one described in murine infection with the pathogen Leishmania major, a kinetoplastid protozoan with a moderately sized genome containing an estimated 8600 genes [45].…”

Major histocompatibility complex class II-associated peptides control the presentation of bacterial superantigens to T cells.

“…*The concentration of peptide used for the blocking study was a suboptimal dose determined from an independent dose-response curve. ~VI3 usage in the hybridomas was determined by flow cytometry and Northern blot analysis and has been detailed elsewhere (16,27; Wen R., et al, manuscript in preparation). IISince hybridoma 1732 is able to respond to a processed peptide derived from SEB, the ability of SEB to block SEB 121-136 recognition was determined on fixed I-Ab-L cells (8).…”

Binding motifs predict major histocompatibility complex class II-restricted epitopes in the Sendai virus M protein

“…We have previously reported the characterization of a large panel of class II-restricted Sendai virus-specific T-cell hybridomas generated from CD4 ϩ T cells recovered from the respiratory tract of C57BL/6 (H-2 b ) mice with primary Sendai virus pneumonia (6). Using vaccinia virus recombinants expressing individual Sendai virus Enders strain proteins, we determined that the Sendai virus-specific T-cell response was directed against both hemagglutinin-neuraminidase (HN) glycoprotein and matrix (M) protein epitopes presented in the context of I-A b .…”

“…B3.3F6 is a Sendai virus M-specific CD4 ϩ T-cell hybridoma which was identified by its reactivity to I-A b -transfected L cells infected with either Sendai virus Enders strain or a vaccinia virus recombinant expressing a full-size cDNA for the Enders strain M gene (6). To identify the epitope recognized by this hybridoma, we first sequenced the M gene cDNA clone in plasmid vector pTF1 by using synthetic oligonucleotide primers and the dideoxynucleotide termination method according to the Sequenase kit protocol (U.S. Biochemical Corp., Cleveland, Ohio).…”

“…Peptides were tested for their ability to stimulate the Mspecific B3.3F6 hybridoma. In addition, we also tested B3.4D4 and B3.7F5, two Sendai virus-specific class II-restricted hybridomas of undetermined protein specificity (6). Hybridoma cells (10 5 ) were cocultured with I-A b -transfected L cells (10 5 ) (25), and individual peptides were added to approximately 60 M in a final volume of 250 l of medium.…”