Protein expression of lymphocytes in HLA-DR transgenic pigs by a proteomic approach.
Matching donor and recipient human leucocyte antigen (HLA-II) could conquer cell-mediated rejection following transplantation. Transgenic pigs carrying HLA genes that "humanize" porcine organs, tissues, and cells were successfully generated. This study further clarifies the effect of HLA-DR transgenes on lymphocyte protein expression, via a proteomic approach. Lymphocytes were isolated from two HLA-DR transgenic pigs and three nontransgenic littermates on 157 d after birth. Soluble protein of 1x10(7) cells was separated using 2-DE. In total, 301 colloidal CBB-stained protein spots detected on all five 2-D gels were quantified. Thirty-three proteins were differentially expressed by a factor of 1.5. These proteins were subsequently identified by MALDI-TOF MS and MALDI-TOF/TOF MS/MS. These proteins were sorted into the following categories: chaperones, T-lymphocyte function, DNA/RNA processing, cytoskeleton-associated proteins, signal transduction, enzymes, and unknown. Previous studies have suggested that some of the identified proteins are associated with lymphocyte activation/proliferation. The identities of the unidentified spots and the systematic effect of these up- and down-regulated proteins on T-cell function in HLA-DR transgenic pigs require further exploration.
Huang
San-Yuan
SY
Division of Biotechnology, Animal Technology Institute Taiwan, Miaoli, Taiwan.
Chen
Yu-Hui
YH
Teng
Shih-Hua
SH
Chen
I-Chung
IC
Ho
Lin-Lin
LL
Tu
Ching-Fu
CF
D000818:Animals; D030801:Animals, Genetically Modified; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D006684:HLA-DR Antigens / Q000235:genetics* / Q000502:physiology; D007150:Immunohistochemistry; D007526:Isoelectric Point; D008214:Lymphocytes / Q000276:immunology*; D008297:Male; D008970:Molecular Weight; D010449:Peptide Mapping; D020543:Proteome / Q000032:analysis* / Q000145:classification; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013552:Swine; D019076:Transgenes
D006684:HLA-DR Antigens; D020543:Proteome