Relationship between Allelic Heterozygosity in BoLA-DRB3 and Proviral Loads in Bovine Leukemia Virus-Infected Cattle

SIMPLE SUMMARY: Bovine leukemia virus (BLV) caused a severe cattle neoplastic disease called enzootic bovine leukosis (EBL). EBL causes significant economic losses in farming by reducing milk production, reproductive performance, and fertility, and through cattle culling or death. The BLV proviral load (PVL) represents the quantity of BLV genome that has integrated into the host’s genome in BLV-infected cells. It has been reported that PVLs differ according to the genetic background of the host, and some studies of BLV-associated host factors have reported on polymorphisms within the bovine major histocompatibility complex (MHC), namely bovine MHC is bovine leukocyte antigen (BoLA-DRB3). However, there is a great diversity in the PVLs associated with carrying various combinations of these alleles, especially for heterozygous alleles. Therefore, this research investigated whether heterogeneity in BoLA-DRB3 allele combinations would affect PVLs during BLV infections in different ages and breeds of cattle in Japan. This is the first report where the association between heterozygous allelic combinations and BLV PVLs phenotypes (HPLs, LPLs) was analyzed. Our findings augment current understanding about the immunological role played by BoLA heterozygosity in BLV-associated PVLs and biocontrol in BLV infections. ABSTRACT: Enzootic bovine leukosis is a lethal neoplastic disease caused by bovine leukemia virus (BLV), belongs to family Retroviridae. The BLV proviral load (PVL) represents the quantity of BLV genome that has integrated into the host’s genome in BLV-infected cells. Bovine leukocyte antigen (BoLA) class II allelic polymorphisms are associated with PVLs in BLV-infected cattle. We sought to identify relationships between BoLA-DRB3 allelic heterozygosity and BLV PVLs among different cattle breeds. Blood samples from 598 BLV-infected cattle were quantified to determine their PVLs by real-time polymerase chain reaction. The results were confirmed by a BLV-enzyme-linked immunosorbent assay. Restriction fragment length polymorphism-polymerase chain reaction identified 22 BoLA-DRB3 alleles. Multivariate negative binomial regression modeling was used to test for associations between BLV PVLs and BoLA-DRB3 alleles. BoLA-DRB3.2*3, *7, *8, *11, *22, *24, and *28 alleles were significantly associated with low PVLs. BoLA-DRB3.2*10 was significantly associated with high PVLs. Some heterozygous allele combinations were associated with low PVLs (*3/*28, *7/*8, *8/*11, *10/*11, and *11/*16); others were associated with high PVLs (*1/*41, *10/*16, *10/*41, *16/*27, and *22/*27). Interestingly, the BoLA-DRB3.2*11 heterozygous allele was always strongly and independently associated with low PVLs. This is the first reported evidence of an association between heterozygous allelic combinations and BLV PVLs.