Journal Article

Preparation and characterization of monoclonal antibodies recognizing two CD4 isotypes of Microminipigs

Hitoshi Kitagawa, Noriaki Hirayama, Masafumi Tanaka, Noriaki Imaeda, Masaki Takasu, Yoshie Kametani, Asuka Miyamoto, Tatsuya Matsubara, Shino Ohshima, Takashi Shiina, Asako Ando, Shingo Suzuki, Atsuko Shigenari, Jerzy K. Kulski

2020

Overview

Cluster of differentiation 4 (CD4) molecule expressed on the leukocytes is known to function as a co-receptor for class II major histocompatibility complex (MHC) binding to T cell receptor (TCR) on helper T cells. We previously identified two CD4 alleles (CD4.A and CD4.B) in a Microminipig population based on nucleotide sequencing and PCR detection of their gene sequences. However, CD4.B protein expression was not examined because of the unavailability of a reactive antibody to a CD4.B epitope. In this study, we have produced two swine-specific monoclonal antibodies (mAbs) against CD4.B molecules, one that recognizes only CD4.B (b1D7) and the other that recognizes both the CD4.A and CD4.B alleles (x1E10) and that can be used to distinguish CD4 T cell subsets by flow cytometry and immunohistochemistry. Using these two mAbs, we identified CD4.A and CD4.B allele-specific proteins on the surface of CD4.A (+/+) and CD4.B (+/+) T cells at a similar level of expression. Moreover, stimulation of peripheral blood mononuclear cells (PBMCs) derived from CD4.A (+/+) and CD4.B (+/+) swine with toxic shock syndrome toxin-1 (TSST-1) in vitro similarly activated both groups of cells that exhibited a slight increase in the CD4/CD8 double positive (DP) cell ratio. A large portion of the DP cells from the allelic CD4.A (+/+) and CD4.B (+/+) groups enhanced the total CD4 and class I swine leukocyte antigen (SLA) expression. The x1E10 mAb delayed and reduced the TSST-1-induced activation of CD4 T cells. Thus, CD4.B appears to be a functional protein whose expression on activated T cells is analogous to CD4.A.

• Publisher: Public Library of Science (PLoS),Public Library of Science
• Subject: Acids / Alleles / Amino Acid Sequence / Animals / Antibodies, Monoclonal / Antibodies, Monoclonal - immunology / Antibody Specificity / Antigens / Biology and Life Sciences / CD4 antigen / CD4 Antigens / CD4 Antigens - analysis / CD4 Antigens - chemistry / CD4 Antigens - immunology / CD8 antigen / CD8 Antigens / CD8 Antigens - analysis / Cell activation / Cell Line, Tumor / Epitopes / Female / Flow cytometry / Gene sequencing / Genotype / Haplotypes / HEK293 Cells / Histocompatibility Antigens Class I / Histocompatibility Antigens Class I - genetics / Histocompatibility Antigens Class I - immunology / Humans / Immunohistochemistry / Isotypes / Laboratory animals / Leukocytes / Leukocytes, Mononuclear / Leukocytes, Mononuclear - immunology / Life sciences / Livestock / Lymphocyte Activation / Lymphocytes / Lymphocytes T / Major histocompatibility complex / Male / Medicine / Medicine and Health Sciences / Mice / Mice, Inbred BALB C / Models, Molecular / Monoclonal antibodies / Nucleotides / Peripheral blood mononuclear cells / Physiological aspects / Population genetics / Product development / Production processes / Protein Conformation / Protein expression / Proteins / Q / R / Receptors / Research and Analysis Methods / Research Article / Science / Septic shock / Sequence Alignment / Sequence Homology, Amino Acid / Signal transduction / Specific Pathogen-Free Organisms / Staphylococcal enterotoxin F / Structure / Swine / Swine, Miniature / Swine, Miniature - genetics / Swine, Miniature - immunology / T cell receptors / T cells / T-cell receptor / Toxic shock syndrome / Toxins / Transfection / Veterinary medicine