Searching for MHC‐restricted anti‐viral antibodies: antibodies recognizing the nucleoprotein of influenza virus dominate the serological response of C57BL/6 mice to syngeneic influenza‐infected cells

Abstract

An attempt has been made to generate monoclonal antibodies which recognize the same target structures on influenza-infected cells as those seen by cytotoxic T lymphocyte (CTL) receptors. Such antibodies, if they mimicked the T cell receptor specificity, would be expected to be both virus specific and restricted in their binding by the major histocompatibility complex (MHC) antigens. Approximately 200 hybridomas from C57BL/6 (H-2b) mice primed and boosted with influenza virus (X-31)-infected EL4 (a C57BL/6 T cell lymphoma) were screened for reactivity on infected and uninfected cells of different MHC haplotypes. Of the 10 hybridoma antibodies which were identified as being reactive with X-31-infected EL4, but not uninfected EL4, all reacted equally well with X-31-infected cells of H-2b, H-2d and H-2k haplotypes, indicating a lack of MHC restriction in their recognition of the infected cells. Unexpectedly, 7 of the 10 monoclonal antibodies were found to react specifically with the purified influenza virus nucleoprotein (NP), a predominant viral antigen in CTL recognition of infected cells. Fluorescence-activated flow cytometry confirmed that these antibodies were able to recognize NP serological determinants on the surface of viable, infected cells, but the anti-NP antibodies were unable to block the lytic activity of an NP-specific CTL clone.

A recombinant antibody with the antigen-specific, major histocompatibility complex-restricted specificity of T cells.

“…4), whereas no effect was observed on two NP50-57-specific, Kk-restricted, T-cell hybridomas, HK9.5-24 and HK9.5-162 (Fig. 4 (12)(13)(14), whereas others have reported that their attempts have failed (15,16). More recently, a few anti-peptide/MHC antibodies have been reported (17)(18)(19).…”

Novel antibodies as anticancer agents

“…Several groups have made deliberate attempts to generate antibodies recognizing T-cell epitopes (Wylie et al, 1982;Froscher and Klinman, 1986;Tamminen et al, 1987;Abastado et al, 1989;Duc et al, 1993;Andersen et al, 1996;Porgador et al, 1997;Reiter et al, 1997;Engberg et al, 1999). In the majority of the examples cited, conventional hybridoma technology was used.…”

Direct selection of a human antibody fragment directed against the tumor T-cell epitope HLA-A1–MAGE-A1 from a nonimmunized phage-Fab library

“…Antibodies that specifically recognize a peptide-MHC complex have already been used to study MHC class I or II antigen presentation (12)(13)(14)(15)(16)(17)(18), to localize and quantify antigen-presenting cells (APC) displaying a T cell epitope (13,(19)(20)(21), specifically mask an autoimmune T cell epitope (22,23), or as a targeting tool in a mouse model (24). However, selecting such reagents remains a difficult task and several failures have been reported (25,26). The available antibodies have been obtained after immunization of mice with recombinant peptide-MHC complexes or peptide-loaded TAP-deficient APC, and recently by selection from phage-antibody libraries made from immunized transgenic mice (27).…”