J Vet Sci.  2018 May;19(3):384-392. 10.4142/jvs.2018.19.3.384.

Analysis of swine leukocyte antigen class I gene profiles and porcine endogenous retrovirus viremia level in a transgenic porcine herd inbred for xenotransplantation research

• MatczyÅ„ska D
• Sypniewski D
• GaÅ‚ka S
• SoÅ‚tysik
• Loch T
• Nowak E
• SmorÄ…g Z
• Bednarek I

• KMID: 2412129
• DOI: http://doi.org/10.4142/jvs.2018.19.3.384

Abstract

Molecular characterization of swine leukocyte antigen (SLA) genes is important for elucidating the immune responses between swine-donor and human-recipient in xenotransplantation. Examination of associations between alleles of SLA class I genes, type of pig genetic modification, porcine endogenous retrovirus (PERV) viral titer, and PERV subtypes may shed light on the nature of xenograft acceptance or rejection and the safety of xenotransplantation. No significant difference in PERV gag RNA level between transgenic and non-transgenic pigs was noted; likewise, the type of applied transgene had no impact on PERV viremia. SLA-1 gene profile type may correspond with PERV level in blood and thereby influence infectiveness. Screening of pigs should provide selection of animals with low PERV expression and exclusion of specimens with PERV-C in the genome due to possible recombination between A and C subtypes, which may lead to autoinfection. Presence of PERV-C integrated in the genome was detected in 31.25% of specimens, but statistically significant increased viremia in specimens with PERV-C was not observed. There is a need for multidirectional molecular characterization (SLA typing, viremia estimation, and PERV subtype screening) of animals intended for xenotransplantation research in the interest of xeno-recipient safety.

Keyword

MHC class II genes; Retroviridae; heterologous transplantation; infection; viremia

MeSH Terms

Alleles
Animals
Endogenous Retroviruses*
Genes, MHC Class I*
Genes, MHC Class II
Genome
Heterografts
Leukocytes*
Mass Screening
Recombination, Genetic
Retroviridae
RNA
Swine*
Transgenes
Transplantation, Heterologous*
Viremia*
RNA

Figure

• 

  Fig. 1 Schematic organization of the SLA class I gene cluster including three major gene groups: class I (1.1 Mbp), class II (0.7 Mbp) (mapped to the chromosome short arm, band 7p1.1), and class III (0.5 Mbp) (on the long arm, band 7q1.1). SLA-1, -2, and -3 genes are indicated in red. SLA, swine leukocyte antigen.

• 

  Fig. 2 A representative swine leukocyte antigen (SLA) class I pattern of an analyzed porcine blood sample. This specimen represents the SLA class I profile, SLA-1ab 2a 3bc (polymerase chain reaction products: 548, 548, 585, 565, and 533 bp, respectively).

• 

  Fig. 3 Percentile share of swine leukocyte antigen (SLA) class I-based groups within the investigated herd. Clustering based on SLA-1 (A), SLA-2 (B), and SLA-3 (C) profiles. Within the investigated herd there were 7 of 8 identifiable combinations of the SLA-1 profile, four (all possible) versions of SLA-2 genes, and 14 of 16 possible combinations of SLA-3. The most often repetitive SLA-1 profile was SLA-1ab, found in 23.5% of the analyzed specimens. More than half of the herd (54.9%) demonstrated a homologous SLA-2 genes profile: SLA-2ab and SLA-3c profiles, together with SLA-3ac, the most abundantly occurring profile among the herd (15.7% each).

• 

  Fig. 4 Average porcine endogenous retrovirus (PERV) viremia within SLA-1 (A), SLA-2 (B), and SLA-3 (C) profile-based groups. Statistically significant differences in PERV gag RNA level were observed only in the SLA-1 profile-based group (A). SLA-1bc group displayed significantly higher viremia than that in the SLA-1a, 1b, 1c, and 1ac profiles. No correlation with PERV viremia and the SLA-2 (B) or SLA-3 profiles (C) was detected. SLA, swine leukocyte antigen.

• 

  Fig. 5 Representative images of multiplex polymerase chain reaction (PCR) results for porcine endogenous retrovirus (PERV)-A, -B, and -C screening. Multiplex PCR with env-specific primers with denaturing polyacrylamide gel electrophoresis was conducted to detect the subtypes of PERV present in genome of pigs. As a result, 3 bands (364, 284, and 270 bp length) were obtained for PERV-C-positive specimens and 2 bands (364 and 270 bp length) for pigs without the PERV-C subtype in their genome. Less than one-third of the herd (31.25%) showed the presence of PERV-C in their genome, and no association with increased viremia was detected.

• 

  Fig. 6 Distribution of particular swine leukocyte antigen (SLA) class I profiles within the assessed herd. Classes were divided into two subgroups: with and without porcine endogenous retrovirus (PERV)-C subtype in their genome (left and right column, respectively, for each hue). In the PERV-C-positive group, the highest occurrences were in the following SLA class I profiles: SLA-1, ab; SLA-2, ab; and SLA-3, c. The SLA-1ab and SLA-3c profiles were present almost twice as often in the PERV-C-positive specimens than in the PERV-c-negative specimens.

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