TCR and CD3 antibody cross‐reactivity in 44 species

Abstract

The production of monoclonal antibodies is very costly, and antibodies are only available for a limited number of species. Until a more cost effective method of antibody production is found, identification of cross-reactive antibodies is an alternative approach that can provide investigators studying immunity in minor species with valuable antibody reagents. Flow cytometry was used to test 21 monoclonal antibodies (mAb), raised against ab and gd T cell receptors and CD3 from human and five animal species, for cross-reactivity in 44 different species including 16 species of nonhuman primates, marsupials, carnivores, lagomorphs, rodents, ruminants, swine, cetacean, horse, birds, a reptile, and fish. Fifteen of the mAbs cross-reacted with orthologous molecules in one or more species. Two antibodies, anti-human TCR gd (B1.1), and anti-human CD3 (SP34) were found to costain in 13 species of nonhuman primates. This study has identified valuable new reagents for studying T cell populations in different animal species and for the first time characterized antibodies useful for studying gd T cell populations in many species of primates. These antibodies may be used for further immunity research in species with less well-characterized immune systems. ' 2007 International Society for Analytical Cytology Key terms monoclonal antibody; T cell receptor; ab; gd; CD3; WC1; cross-reactivity; primate; mammal; carnivore AN interesting finding to emerge from T cell research is the disparity between peripheral blood gd T cell populations in certain animals. Percentages of gd T lymphocytes in peripheral blood vary so greatly between different species that species can be classified as gd high or low (1). gd low species include humans and mice (2-5%) (2,3) and gd high species include chickens (15%) (4), swine (24%) (5), and cattle (20-40%) (6).The high concentration of gd T cells in ruminants and swine is attributable to the presence of a subpopulation of gd T cells that express the workshop cluster 1 (WC1) molecule in ruminants and the orthologue in swine. WC1 and orthologues have only been identified in artiodactyls including ruminants, swine, and camelids (7,8). The current data suggest this is a unique population of gd T cells that has evolved in artiodactyls. The concentration of WC1-gd T cells identified in ruminants and swine is similar to the concentration seen in humans and mice (9,10). Analysis of gd T cells in chickens has not revealed an explanation for the high concentration of gd T cells. The lack of reagents has thus far limited analysis of the composition and function of gd T cells in additional species. Identification of cross reactive mAbs would facilitate the conduct of further studies and an opportunity to delineate any functional differences in populations of gd T cells present in species with high and low concentrations of gd T cells in peripheral blood.Comparative studies with mAbs developed against major histocompatibility and leukocyte differentiation antigens in humans and other species have demonstr...

Ontogenic development of kidney, thymus and spleen and phenotypic expression of CD3 and CD4 receptors on the lymphocytes of cobia (Rachycentroncanadum )

“…The use of cross-reactivity is only possible because molecules present on T cell receptors of some species, are conserved during evolution (Romano et al 2004, Conrad et al 2007, Batista et al 2014. This work has demonstrated the usefulness of classical histology and immunohistochemistry, which are accessible and technically simple tools, to evaluate the ontogenic development of the cobia and contribute o the cultivation and future research of this fish.…”

Complex mixtures, complex responses: Assessing pharmaceutical mixtures using field and laboratory approaches

“…Limited reagents exist to study teleost immune systems, and none of the existing reagents was raised against fathead minnow epitopes. Fortunately, some proteins, such as a CD3 (a portion of the T-lymphocyte's receptor), are conserved across fish species, which allowed the use of cross-reactive antibodies obtained against one species to detect that same protein in other species [41,53]. The proportion of B lymphocytes did not change in any of the treated groups relative to the control group, whereas both the antidepressant and tramadol groups contained increased proportions of T lymphocytes.…”

Fat-associated lymphoid cluster in Cyprinus carpio: Characterisation and its relation with peritoneal haemangiosarcoma

“…The slides were blocked for 60 min with blocking solution (0.25% albumin in PBS) and then incubated at 37 C for 60 min with primary antibodies (Abcam™). In the market, no specific antibodies for all of the fish surface markers assessed in this study exist; however, in the case of polyclonal antibodies, cross-reactivity between some vertebrates was demonstrated [4,12,13,51,57,67]. The polyclonal anti-c-kit antibodies possess variations in sensitivity, but do not stain cells other than those expressing c-kit [57,67].…”

“…Anti-CD3 (ab113628 polyclonal Ab produced in rabbit) is a synthetic peptide corresponding to the extracellular domain of human CD3 with a cross-reactivity between mouse, rat and human, and contain one ITAM domain. However, some trademarks of antibodies identified epitopes conserved on orthologous molecules in one or more species [12,13]. This antibody was used at a dilution of 1:100.…”