PMID- 15303984
OWN - NLM
STAT- MEDLINE
DCOM- 20041201
LR  - 20040812
IS  - 0908-665X (Print)
IS  - 0908-665X (Linking)
VI  - 11
IP  - 5
DP  - 2004 Sep
TI  - Rapid detection of human HLA transgenes in pigs by fluorescence in situ 
      hybridization (FISH) for adjuvant study of human xenotransplantation.
PG  - 471-5
AB  - OBJECTIVES: We have recently generated several lines of transgenic pigs for 
      HLA-DP and -DQ to elucidate the role of HLA-II antigens in the modulation of 
      cell-mediated rejection of xenotransplantation. Using fluorescence in situ 
      hybridization (FISH) analysis, the aim of this study was to determine integration 
      sites and to test zygosity of these transgenes in the piglets after cross mating. 
      METHODS: Blood lymphocytes of transgenic pigs for HLA-DP and -DQ were collected 
      and cultured. Chromosome spreads were prepared by standard methodology. Gene 
      constructs of HLA-DP A1+B1, -DQ A1 & B1 were labeled with fluorescein 
      isothiocyanate or Texas Red by nick-translation. Hybridization was based on a 
      standard FISH protocol. RESULTS: FISH analysis revealed that the HLA-DP probe 
      hybridized to porcine chromosome 6, while both HLA-DQ A1 and B1 probes hybridized 
      to porcine chromosome 11 at the same site. There was no cross-hybridization of 
      HLA transgenes to the swine leukocyte antigen complex. Mosaic integration of 
      HLA-DQ transgenes in the genome of F0, but full penetrance in F1 after selective 
      breeding was observed. Both HLA-DP and HLA-DQ lines were determined to be 
      heterozygous at the integration site. CONCLUSION: By FISH, we have detected 
      specific integration sites of the HLA-DP and -DQ transgenes in pig genome and 
      determined mosaic levels and zygosity types of these transgenes. We conclude that 
      FISH is both sensitive and labor-efficient in confirming and differentiating 
      transgenic pigs for multiple rejection-regulatory genes by visualizing individual 
      integration sites in chromosomes or interphase nuclei.
CI  - Copyright 2004 Blackwell Munksgaard
FAU - Wang, Bao-Tyan
AU  - Wang BT
AD  - Department of Medical Research, Changhua Christian Hospital, Changhua, Taiwan.
FAU - Tu, Ching-Fu
AU  - Tu CF
FAU - Hsieh, Lie-Jiau
AU  - Hsieh LJ
FAU - Tai, Hao-Chih
AU  - Tai HC
FAU - Chiu, Ya-Lan
AU  - Chiu YL
FAU - Lee, Jang-Ming
AU  - Lee JM
FAU - Kuo, Shou-Jen
AU  - Kuo SJ
FAU - Tsuji, Kimiyoshi
AU  - Tsuji K
FAU - Lee, Chun-Jean
AU  - Lee CJ
LA  - eng
PT  - Journal Article
PL  - Denmark
TA  - Xenotransplantation
JT  - Xenotransplantation
JID - 9438793
RN  - 0 (Antibodies, Heterophile)
RN  - 0 (HLA Antigens)
RN  - 0 (HLA-DP Antigens)
RN  - 0 (HLA-DQ Antigens)
SB  - IM
MH  - Animals
MH  - *Animals, Genetically Modified
MH  - Antibodies, Heterophile/blood
MH  - Graft Rejection/immunology
MH  - HLA Antigens/analysis/*genetics
MH  - HLA-DP Antigens/blood/genetics
MH  - HLA-DQ Antigens/blood/genetics
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Swine
MH  - Transplantation, Heterologous/*immunology
EDAT- 2004/08/12 05:00
MHDA- 2004/12/16 09:00
CRDT- 2004/08/12 05:00
PHST- 2004/08/12 05:00 [pubmed]
PHST- 2004/12/16 09:00 [medline]
PHST- 2004/08/12 05:00 [entrez]
AID - XEN160 [pii]
AID - 10.1111/j.1399-3089.2004.00160.x [doi]
PST - ppublish
SO  - Xenotransplantation. 2004 Sep;11(5):471-5. doi: 10.1111/j.1399-3089.2004.00160.x.