A novel monoclonal antibody specific for canine CD25 (P4A10): Selection and evaluation of canine Tregs

V. Kraig Abrams; Billanna Hwang; Marina Lesnikova; M. John Gass; Elizabeth Wayner; Cristina Castilla-Llorente; George E. Georges; Richard A. Nash

doi:10.1016/j.vetimm.2009.12.006

A novel monoclonal antibody specific for canine CD25 (P4A10): Selection and evaluation of canine Tregs

V. Kraig Abrams^*, Billanna Hwang, Marina Lesnikova, M. John Gass, Elizabeth Wayner, Cristina Castilla-Llorente, George E. Georges, Richard A. Nash

^*Corresponding author for this work

Medicine, Hematology Oncology Division

Research output: Contribution to journal › Article › peer-review

24 Scopus citations

Abstract

A monoclonal antibody (mAb), P4A10, was made to the canine interleukin-2 receptor alpha chain (IL-2Rα; p55; Tac antigen; CD25) to facilitate studies of canine regulatory T-cells (Treg). By non-reduced Western blot, P4A10 bound to a 55kDa protein, the size of human IL-2Rα. In flow cytometry assays, it reacted with a minor population of circulating dog CD3⁺CD4⁺ T-cells and the majority (>60%) of in vitro PMA-Ionomycin (PMA-IO)-activated canine CD3⁺ T-cells. P4A10 recognized a hematopoietic cell population enriched for FoxP3+ cells as measured by flow cytometry. The P4A10-selected fractions of T-cells had significantly increased copy numbers of CD25, FoxP3, IL-10, and TGFβ as detected by RT-PCR (reverse transcriptase-PCR) compared to the negative fractions. The P4A10-selected cells inhibited ³H (tritiated) thymidine incorporation in a mixed leukocyte reaction (MLR) containing responders of the same origin. P4A10-selected T-cells from fresh peripheral blood mononuclear cells had less FoxP3 (p=0.07) by qRT-PCR (quantitative RT-PCR) and were less suppressive (p=0.01) than in vitro alloantigen-activated Treg. The mAb P4A10 is specific for canine CD25 and can be used to facilitate studies of CD25+FoxP3+ Treg in this clinically relevant large animal model.

Original language	English (US)
Pages (from-to)	257-265
Number of pages	9
Journal	Veterinary Immunology and Immunopathology
Volume	135
Issue number	3-4
DOIs	https://doi.org/10.1016/j.vetimm.2009.12.006
State	Published - Jun 2010

Funding

Supported in part by NIH grants AI69879 , DK42716 and CA15704 from the NIH, DHHS, Bethesda, MD. The authors of this report have no financial or personal relationships with other people or organizations that could inappropriately influence (bias) this work. The work was funded through the National Institutes of Health and the authors were solely responsible for the study design, the collection, analysis and interpretation of data as well as writing the manuscript.

Keywords

Cd25
Dog
Monoclonal antibody
Regulatory T-cells

ASJC Scopus subject areas

General Veterinary
Immunology

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10.1016/j.vetimm.2009.12.006

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title = "A novel monoclonal antibody specific for canine CD25 (P4A10): Selection and evaluation of canine Tregs",

abstract = "A monoclonal antibody (mAb), P4A10, was made to the canine interleukin-2 receptor alpha chain (IL-2Rα; p55; Tac antigen; CD25) to facilitate studies of canine regulatory T-cells (Treg). By non-reduced Western blot, P4A10 bound to a 55kDa protein, the size of human IL-2Rα. In flow cytometry assays, it reacted with a minor population of circulating dog CD3+CD4+ T-cells and the majority (>60%) of in vitro PMA-Ionomycin (PMA-IO)-activated canine CD3+ T-cells. P4A10 recognized a hematopoietic cell population enriched for FoxP3+ cells as measured by flow cytometry. The P4A10-selected fractions of T-cells had significantly increased copy numbers of CD25, FoxP3, IL-10, and TGFβ as detected by RT-PCR (reverse transcriptase-PCR) compared to the negative fractions. The P4A10-selected cells inhibited 3H (tritiated) thymidine incorporation in a mixed leukocyte reaction (MLR) containing responders of the same origin. P4A10-selected T-cells from fresh peripheral blood mononuclear cells had less FoxP3 (p=0.07) by qRT-PCR (quantitative RT-PCR) and were less suppressive (p=0.01) than in vitro alloantigen-activated Treg. The mAb P4A10 is specific for canine CD25 and can be used to facilitate studies of CD25+FoxP3+ Treg in this clinically relevant large animal model.",

keywords = "Cd25, Dog, Monoclonal antibody, Regulatory T-cells",

author = "Abrams, {V. Kraig} and Billanna Hwang and Marina Lesnikova and Gass, {M. John} and Elizabeth Wayner and Cristina Castilla-Llorente and Georges, {George E.} and Nash, {Richard A.}",

note = "Funding Information: Supported in part by NIH grants AI69879 , DK42716 and CA15704 from the NIH, DHHS, Bethesda, MD. Funding Information: The authors of this report have no financial or personal relationships with other people or organizations that could inappropriately influence (bias) this work. The work was funded through the National Institutes of Health and the authors were solely responsible for the study design, the collection, analysis and interpretation of data as well as writing the manuscript. ",

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AU - Castilla-Llorente, Cristina

AU - Georges, George E.

AU - Nash, Richard A.

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A novel monoclonal antibody specific for canine CD25 (P4A10): Selection and evaluation of canine Tregs

Abstract

Funding

Keywords

ASJC Scopus subject areas

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