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Evidence for antigen recognition by nonspecific cytotoxic cells: initiation of 3H-thymidine uptake following stimulation by a protozoan parasite and homologous cognate synthetic peptide.

Identifieur interne : 003A71 ( Main/Exploration ); précédent : 003A70; suivant : 003A72

Evidence for antigen recognition by nonspecific cytotoxic cells: initiation of 3H-thymidine uptake following stimulation by a protozoan parasite and homologous cognate synthetic peptide.

Auteurs : Donald L. Evans [États-Unis] ; John H. Leary Jr. Iii [États-Unis] ; Praveen Nadella [États-Unis] ; Liliana Jaso-Friedmann [États-Unis]

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RBID : ISTEX:E7C4614172B34BCF1584FF6020F4C83710F29BB5

English descriptors

Abstract

Abstract: Catfish nonspecific cytotoxic cells bind to and lyse certain protozoan parasites and tumor cells. Target cell binding is facilitated by recognition of (minimally) one antigenic determinant. Binding to this determinant initiates multiple signalling pathways in NCC including proto-oncogene kinase phosphorylation, regulation of phosphatase activity and increased membrane receptor expression. In the present study, highly purified NCC were activated in vitro with the protozoan parasite Tetrahymena pyriformis, with a multiple antigenic peptide (MAP) composed of the cognate antigenic determinant of this parasite (i.e. natural killer target antigen/NKTag) and NCC were activated with a monoclonal antibody specific for the NCC receptor which binds NKTag. NCC were purified by Percoll density gradients and negative selection by panning (2×) over anti-sIg specific mab 9E1. In 5 day proliferation experiments, treatment of NCC with immobilized Tetrahymena initiated a significant increase in uptake of tritiated thymidine. This appeared to be a primary response in that NCC from in vivo parasite primed catfish did not have secondary-like proliferation responses. Stimulation of NCC with immobilized synthetic peptides composed of the cognate antigenic determinant of this parasite (i.e. MAP) also caused significant increased uptake of tritiated thymidine. An indication that NCC recognize a specific antigenic determinant was that sMAP (i.e. peptides composed of the same amino acids as MAP but in a scrambled sequence) failed to increase incorporation. Similar to the MAP results, mab 5C6 binding to NCC also caused increased thymidine uptake. To determine if an IL-2 cosignal was required to achieve optimum activation responses by NCC, different concentrations of human recombinant IL-2 (rHuIL-2) were tested individually or as costimulants. Co-treatment of NCC with rHuIL-2 and any of the three stimuli (parasite, MAP, mab 5C6) did not produce increased proliferation of NCC. These studies demonstrated that NCC specifically recognize an antigenic determinant on protozoan parasites and binding to this antigen produces an activation signal that may have important consequences for elicitation of innate immunity.

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DOI: 10.1016/S0145-305X(97)00046-3


Affiliations:


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Le document en format XML

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<term>Assay</term>
<term>Biotinylated peptide</term>
<term>Cat_sh</term>
<term>Channel cat_sh</term>
<term>Cognate</term>
<term>Color analysis</term>
<term>Cytokine</term>
<term>Cytometry</term>
<term>Cytotoxic</term>
<term>Cytotoxic cells</term>
<term>Cytotoxicity</term>
<term>Density gradient</term>
<term>Determinant</term>
<term>Elsevier science</term>
<term>Fish biol</term>
<term>Further puri_ed</term>
<term>Ictalurus punctatus</term>
<term>Immunol</term>
<term>Isotype</term>
<term>Isotype control</term>
<term>Killer cells</term>
<term>Lanier</term>
<term>Leary</term>
<term>Liferation responses</term>
<term>Medical microbiology</term>
<term>Microtiter plates</term>
<term>Monoclonal</term>
<term>Monoclonal antibody</term>
<term>Moretta</term>
<term>Multiple antigenic peptide</term>
<term>Natural killer</term>
<term>Negative selection</term>
<term>Nktag</term>
<term>Nonspeci_c</term>
<term>Nonspeci_c cytotoxic cells</term>
<term>Parasite</term>
<term>Peptide</term>
<term>Percoll</term>
<term>Peripheral blood</term>
<term>Present study</term>
<term>Proliferation</term>
<term>Proliferation assays</term>
<term>Proliferation experiment</term>
<term>Proliferation experiments</term>
<term>Proliferation response</term>
<term>Proliferation responses</term>
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<term>Receptor</term>
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<term>Signi_cant</term>
<term>Signi_cant increase</term>
<term>Speci_c</term>
<term>Spleen</term>
<term>Spleen cells</term>
<term>Standard techniques</term>
<term>Synthetic peptides</term>
<term>Target cells</term>
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<term>Tritiated thymidine</term>
<term>Tritiated thymidine uptake</term>
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<term>Unfractionated spleen cells</term>
<term>Uptake</term>
<term>Veterinary medicine</term>
<term>Wide variety</term>
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<div type="abstract" xml:lang="en">Abstract: Catfish nonspecific cytotoxic cells bind to and lyse certain protozoan parasites and tumor cells. Target cell binding is facilitated by recognition of (minimally) one antigenic determinant. Binding to this determinant initiates multiple signalling pathways in NCC including proto-oncogene kinase phosphorylation, regulation of phosphatase activity and increased membrane receptor expression. In the present study, highly purified NCC were activated in vitro with the protozoan parasite Tetrahymena pyriformis, with a multiple antigenic peptide (MAP) composed of the cognate antigenic determinant of this parasite (i.e. natural killer target antigen/NKTag) and NCC were activated with a monoclonal antibody specific for the NCC receptor which binds NKTag. NCC were purified by Percoll density gradients and negative selection by panning (2×) over anti-sIg specific mab 9E1. In 5 day proliferation experiments, treatment of NCC with immobilized Tetrahymena initiated a significant increase in uptake of tritiated thymidine. This appeared to be a primary response in that NCC from in vivo parasite primed catfish did not have secondary-like proliferation responses. Stimulation of NCC with immobilized synthetic peptides composed of the cognate antigenic determinant of this parasite (i.e. MAP) also caused significant increased uptake of tritiated thymidine. An indication that NCC recognize a specific antigenic determinant was that sMAP (i.e. peptides composed of the same amino acids as MAP but in a scrambled sequence) failed to increase incorporation. Similar to the MAP results, mab 5C6 binding to NCC also caused increased thymidine uptake. To determine if an IL-2 cosignal was required to achieve optimum activation responses by NCC, different concentrations of human recombinant IL-2 (rHuIL-2) were tested individually or as costimulants. Co-treatment of NCC with rHuIL-2 and any of the three stimuli (parasite, MAP, mab 5C6) did not produce increased proliferation of NCC. These studies demonstrated that NCC specifically recognize an antigenic determinant on protozoan parasites and binding to this antigen produces an activation signal that may have important consequences for elicitation of innate immunity.</div>
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