Development of an Immortalized Porcine Fibroblast Cell Panel With Different Swine Leukocyte Antigen Genotypes.

Quy Van Chanh Le,Munjeong Choi,Seungyeon Youk,Chak-Sum Ho,Hyoim Jeon,Chankyu Park,Won-Il Kim

doi:10.3389/fgene.2022.815328

Abstract

Immortalized cell lines are valuable resources to expand the molecular characterization of major histocompatibility complex genes and their presented antigens. We generated a panel of immortalized cell lines by transfecting human telomerase reverse transcriptase (hTERT) into primary fibroblast cells prepared from ear, fetal, and lung tissues of 10 pigs from five breeds and successfully cultured them for 30–45 passages. The cell growth characteristic of the immortalized fibroblasts was similar to that of primary fibroblast, which was unable to form colonies on soft agar. The genotypes of major swine leukocyte antigen (SLA) genes, including three classical class I (SLA-1, -2, and -3) and three class II genes (DQB1, DRB1, and DQA), were determined using high-resolution typing. A total of 58 alleles, including a novel allele for SLA-2, were identified. Each cell line was unique. A cell line derived from a National Institutes of Health miniature pig was homozygous across the six major SLA genes. The expression levels of SLA classical class I genes varied among the cell lines and were slightly upregulated in the immortalized compared to the primary cells based on semiquantitative reverse transcription polymerase chain reaction. The immortalized porcine fibroblast cell lines with diverse SLA haplotypes that were developed in this study have potential to be applied in studies regarding the molecular characteristics and genetic structure of SLA genes and epitope–major histocompatibility complex interactions in pigs.

Highlights

Antigen presentation by major histocompatibility complex (MHC) class I and II molecules is an essential process in adaptive immune interactions that triggers T cell proliferation and antigenspecific cellular immune responses (Wieczorek et al, 2017)
A total of 16, 13, 8, 6, 8, and 7 alleles were identified for swine leukocyte antigen (SLA)-1, SLA-2, SLA-3, SLA-DQA, SLADQB1, and SLA-DRB1, respectively
Epstein–Barr virus-transformed B-lymphoblastoid cell lines and extensive curation have been important developments in the molecular characterization of MHC genes and MHC-antigen interactions related to HLA (Yang et al, 1989)

Summary

Introduction

Antigen presentation by major histocompatibility complex (MHC) class I and II molecules is an essential process in adaptive immune interactions that triggers T cell proliferation and antigenspecific cellular immune responses (Wieczorek et al, 2017). MHC class I molecules are expressed on the surface of all nucleated cells and interact with cytotoxic T cells (CTL) to trigger immune responses against intracellular pathogenic peptides, derived from viral proteins and tumor cells (Kaufman et al, 1990; Sommer, 2005). A single MHC molecule can bind to multiple peptides and a peptide could be presented by several different MHC molecules (Van Hateren et al, 2010; Eisen et al, 2012) This increases the potential diversity of antigenic peptides repertoire presented to T cells from endogenous and exogenous environments

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