The antibody response to myoglobin is independent of the immunized species. Analysis in terms of replacements in the antigenic sites and in environmental residues of the cross-reactions of fifteen myoglobins with sperm-whale myoglobin antisera raised in different species

Abstract

The recent determination of the entire antigenic structure of sperm-whale myoglobin with rabbit and goat antisera has permitted the examination of whether the antigenic structure recognized by antibodies depends on the species in which the antisera are raised. Also, by knowledge of the antigenic structure, the molecular factors that determine and influence antigenicity can be better understood in terms of the effects of amino acid substitutions occurring in the antigenic sites and in the environmental residues of the sites. In the present work, the myoglobins from finback whale, killer whale, horse, chimpanzee, sheep, goat, bovine, echidna, viscacha, rabbit, dog, cape fox, mouse and chicken were examined for their ability to cross-react with antisera to sperm-whale myoglobin. By immunoadsorbent titration studies with radioiodinated antibodies, each of these myoglobins was able to bind antibodies to sperm-whale myoglobin raised in goat, rabbit, chicken, cat, pig and outbred mouse. It was found that the extent of cross-reaction of a given myoglobin was not dependent on the species in which the antisera were raised. This indicated that the antibody response to sperm-whale myoglobin (i.e. its antigenic structure) is independent of the species in which the antisera are raised and is not directed to regions of sequence differences between the injected myoglobin and the myoglobin of the immunized host. Indeed, in each antiserum from a given species examined, that antiserum reacted with the myoglobin of that species. The extent of this auto-reactivity for a given myoglobin was comparable with the general extent of cross-reactivity shown by that myoglobin with antisera raised in other species. The cross-reactivities and auto-reactivities (both of which are of similar extents for a given myoglobin) can be reasonably rationalized in terms of the effects of amino acid substitutions within the antigenic sites and within the residues close to these sites. These findings confirm that the antigenicity of the sites is inherent in their three-dimensional locations.

Feasibility of a Multiplex Flow Cytometric Bead Immunoassay for Detection of Anti-Epoetin Alfa Antibodies

“…A second donor (donor 63) had a noteworthy reactivity to sperm whale myoglobin. This may reflect a bona fide cross-reactive antibody, since humans are known to have circulating anti-myoglobin antibodies (8,9), and cross-reactivity between anti-myoglobin antibodies and myoglobins from a variety of species is not unusual (25). Additional experiments in this validation exercise included serial dilution of the human positive control serum (Fig.…”

Antigenic structures of proteins. Their determination has revealed important aspects of immune recognition and generated strategies for synthetic mimicking of protein binding sites

“…We have found that the same five antigenic sites on sperm-whale Mb are recognized by rabbit and goat antisera [l] and by chicken, cat, pig, and mouse antisera [46]. Also, rabbit, goat, cat, and mouse antisera recognize the same three antigenic sites on lysozyme [4, 951.…”

“…Recently, we studied the immunochemical cross-reaction of 15 myoglobins at the antibody level in several outbred species of animals 1461 and in selected inbred mouse strains both at the antibody level 1521 and by the T-cell proliferative response [114]. The findings have shown that the immunochemical cross-reactions can be reasonably rationalized on the basis of amio acid substitutions occurring in the residues of the antigenic sites and in the environmental (nearest-neighbour) residues surrounding the sites [46]. Frequently, one-removed or even more distant residues from the sites exerted some effects.…”

Segment α 182‐198 of Torpedo californica acetylcholine receptor contains a second toxin‐binding region and binds anti‐receptor antibodies

“…Also, the binding to AChR peptide of antibodies against unrelated proteins served as additional controls. Antibody binding was determined by a double-antibody assay [28] using 1251-labelled rabbit and mouse IgG to monitor the binding of unlabelled mouse anti-AChR antibodies.…”