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Bovine Leukemia virus

Table of Contents
  1. General Information
    1. NCBI Taxonomy ID
    2. Introduction
  2. Vaccine Related Pathogen Genes
    1. env (Protective antigen)
    2. Env/Rex/Tax (Protective antigen)
    3. gag (Protective antigen)
  3. Vaccine Information
    1. Bovine leukemia virus DNA vaccine encoding Env
    2. Bovine Leukemia Virus Vaccine pBLV6073DX
  4. References
I. General Information
1. NCBI Taxonomy ID:
11901
2. Introduction
Bovine leukemia virus (BLV) is a bovine virus closely related to HTLV-I, a human tumour virus. BLV is a retrovirus which integrates a DNA intermediate as a provirus into the DNA of B-lymphocytes of blood and milk. It contains an oncogene coding for a protein called Tax. Nevertheless in its natural host the cattle leukemia is rare. Because the oncogenic properties of the virus were discovered early, a search for evidence of pathogenicity humans started soon after discovery. Mostly farm workers drinking raw milk were tested for disease, especially for leukemia. But neither leukemia nor other signs of infection could be detected. So many in many states it was not tried to get rid of this infection. Testing strategies have recently changed since the virus was first detected in Cows.

Many potential routes of BLV transmission exist. Transmission through procedures that transmit blood between animals such as gouge dehorning, vaccination and ear tagging with instruments or needles that are not changed or disinfected between animals is a significant means of BLV spread. Rectal palpation with common sleeves poses a risk that is increased by inexperience and increased frequency of palpation. Transmission via colostrum, milk, and in utero exposure is generally considered to account for a relatively small proportion of infections. Embryo transfer and artificial insemination also account for a small number of new infections as long as common equipment and/or palpation sleeves are not used. While transmission has been documented via blood feeding insects, the significance of this risk is unclear. The bottom line appears to be that transmission relies primarily on the transfer of infected lymphocytes from one animal to the next and that BLV positive animals with lymphocytosis are more likely to provide a source for infection.

In general BLV causes only a benign mononucleosis-like disease in cattle. Only some animals later develop a B-cell leukemia called enzootic bovine leukosis. Under natural conditions the disease is transmitted mainly by milk to the calf. Infected lymphocytes transmit the disease too. So for artificial infection infected cells are used or the more stable and even heat resistant DNA. Virus particles are difficult to detect and not used for transmission of infection. It is possible that a natural virus reservoir exists in the water buffalo (Wiki: Bovine leukemia virus).
1. env
  • Gene Name : env
  • Sequence Strain (Species/Organism) : Bovine Leukemia virus
  • NCBI Protein GI : BAV53924
  • Taxonomy ID : 11901
  • Protein Name : gp51
  • Protein pI : 6.33
  • Protein Weight : 17528.5
  • Protein Length : 212
  • Protein Note : genotype: 2
  • Protein Sequence : Show Sequence 
    >BAV53924.1 gp51, partial [Bovine leukemia virus]
IYWPPPQGRRRFGARAMVTYDCEPRCPYVGADRFDCPHWDNASQADQGSFYVDHQILFLHLKQCHGIFTL
TWKIWGYDPLITFSLHKIPDPPQPDFPQLNSDWVPSVRSWALLLNQTARAFPDCAICWEPSPPWAPEILV
YNKTISSSGPGLALPDAQ
  • Molecule Role : Protective antigen
  • Molecule Role Annotation : (Kabeya et al., 1996)
2. Env/Rex/Tax
  • Gene Name : Env/Rex/Tax
  • Sequence Strain (Species/Organism) : Bovine leukemia virus
  • VO ID : VO_0011051
  • NCBI Gene ID : 2760849
  • NCBI Protein GI : 9626230
  • Locus Tag : BLVgp05
  • Genbank Accession : AF033818
  • Protein Accession : NP_056898
  • Taxonomy ID : 11901
  • Gene Starting Position : 4614
  • Gene Ending Position : 7460
  • Gene Strand (Orientation) : +
  • Protein Name : p18
  • Protein pI : 10.96
  • Protein Weight : 16183.28
  • Protein Length : 156
  • DNA Sequence : Show Sequence 
    >NC_001414.1:4614-7460 Bovine leukemia virus, complete genome
AATGCCCAAAGAACGACGGTCCCGAAGACGCCCACAACCGATCATCAGATGGGTAAGTCTCACTCTTACT
CTCCTCGCTCTCTGTCAGCCCATCCAGACTTGGAGATGCTCCCTGTCCCTAGGAAATCAACAATGGATGA
CAACATATAACCAAGAGGCAAAATTTTCCATCTCCATTGACCAAATATTAGAGGCTCACAATCAATCACC
TTTCTGTCCCAGGTCTCCCAGATACACCTTGGACTTCGTAAATGGTTATCCTAAGATCTATTGGCCCCCC
CCACAAGGGCGGCGCCGGTTTGGAGCCAGGGCCATGGTCACATATGATTGCGAGCCCCGATGCCCTTATG
TGGGGGCAGATCACTTCGACTGCCCCCACTGGGACAATGCCTCCCAGGCCGATCAAGGGTCCTTTTATGT
CAATCATCAGATTTTATTCCTGCATCTCAAACAATGTCATGGAATTTTCACTCTAACCTGGGAAATATGG
GGATATGATCCCCTGATCACCTTTTCTTTACATAAAATTCCTGATCCCCCTCAACCCGACTTCCCTCAGC
TGAACAGTGACTGGGTTCCCTCTGTCAGATCATGGGCCCTGCTTTTAAATCAAACGGCACGGGCCTTCCC
AGACTGTGCTATATGTTGGGAACCTTCCCCTCCCTGGGCTCCCGAAATATTAGTATATAACAAAACCATC
TCCGGCTCTGGACCCGGTCTCGCCCTCCCGGACGCCCAAATCTTCTGGGTCAACACGTCCTTGTTTAACA
CCACCCAAGGATGGCACCACCCTTCCCAGAGGTTGTTGTTCAATGTTTCTCAAGGCAACGCCTTATTATT
ACCCCCTATCTCCCTGGTTAATCTCTCTACGGTTTCCTCCGCCCCTCCTACCCGGGTCAGACGCAGTCCC
GTCGCAGCCCTGACCTTAGGCCTAGCCCTGTCAGTGGGGCTCACTGGAATTAATGTAGCCGTGTCTGCCC
TTAGCCATCAGAGACTCACCTCCCTGATCCACGTTCTGGAGCAAGATCAGCAACGCTTGATCACAGCAAT
TAACCAGACCCATTATAATTTGCTTAATGTGGCCTCTGTGGTCGCCCAGAACCGACGGGGGCTTGATTGG
TTGTACATCCGGCTGGGTTTTCAAAGCCTATGTCCCACAATCAATGAACCTTGCTGTTTCCTACGCATTC
AAAATGACTCCATTATCCGCCTCGGTGATCTCCAGCCTCTCTCGCAAAGAGTCTCTACAGACTGGCAGTG
GCCCTGGAATTGGGATCTGGGGCTCACCGCCTGGGTGCGAGAAACCATTCATTCTGTTCTAAGCCTATTC
CTATTAGCCCTTTTTTTGCTCTTCTTGGCCCCCTGCCTGATAAAATGCTTGACCTCTCGCCTTTTAAAAC
TCCTCCGGCAGGCTCCCCACTTCCCTGAAATCTCCTTCCCCCCTAAACCCGATTCTGATTATCAGGCCTT
GCTACCGTCAGCACCAGAGATCTACTCTCACCTCTCCCCCACCAAACCCGATTACATCAACCTTCGACCC
TGCCCTTGACACCCCCGTGTTTCACGCACCCTCAGGCTGTGGTGGGGCACTGGCTTAGTGGAATAGTCAG
TGTACCATCACAAGCCTCTTCTTGCTGCCAGCACCGAGTTCGAACACAGCCCTACCCTGAGCCTCTCTGA
GTACATGACTGAGTGTAGCGCAGAGAGGTTGTCGCTTCTGCGTGTCACTCAGTCATTTTTTATAGCCGAT
TGGGGTTCGCGCCCTCCCATTGCCTGTGACACGGTTAAGACCTCTCTCACTTCTGCTTCACCATCCCCCT
GCCAGCGTTGGTCTAGTGGAAAGAACTAACGCTGACGGGGGCGATTTCTTGCAGCTGTGCTAAGCGAGAG
GCTCTGGTGCTGGGGATAAGATGCGGCCCCTAGCACCACAGTCTCTGCGCCTTTTGGGTTCGAATCTTCC
CCATGCAGCTTCCGCTTTTTACGCCCTGTTGCACACCCTTTCTAGAGATACCTGAAAATCTCAGCTCGCA
CCCCAAGGAAGGTTGTGGCTCAGAGGTTAAAATAGCTCGGACCGCAACCTCCCTTTCTTTTTATTCCACC
CTCGCAAGGCCCCGGGTTCTGGGCCCCCTAACGGAGGTTCAGAATTTCCTCTACTAGGGGATGCTCAGGT
CCAAGTGTGCACAATATCTCTTCCAAAAGGTCCTGATGAACATCTTCCCATGTAACAAGCCCCAGCAGAG
ACATTCCAGCCACATCCAGCAGCATTTGGGCCGCCTTCTCTAACAGTGCCCATAAAGTCCCTTCCGTTTC
CACAACGGCTGCCTCTGCATCTTCTATTTCCACCTCGGCACCGACTCCCCCGCCGAGCCCTTCAAGCTCT
TCGGGATCCATTACCTGATAACGACAAAATTATTTCTTGTCTTTTAAGCAAGTGTTGTTGGTTGGGGGCC
CCACTCTCTACATGCCTGCCCGGCCCTGGTTTTGTCCAATGATGTCACCATCGATGCCTGGTGCCCCCTC
TGCGGGCCCCATGAACGACTCCAATTCGAAAGGATCGACACCACGCTCACCTGCGAGACCCACCGTATCA
CCTGGACCGCCGATGGACGACCTTTTGGCCTCAATGGAACATTGTTCCCTCGACTGCATGTCTCCGAGAC
CCGCCCCCAAGGGCCCCGACGACTCTGGATCAACTGCCCCCTTCCGGCCGTTCGCGCTCAGCCCGGCCCG
GTTTCACTTTCCCCCTTCGAGCAGTCCCCCTTCCAGCCCTACCAATGCCAATTGCCCTCGGCCTCTAGCG
ACGGTTGCCCCATCATCGGGCACGGCCTTCTTCCCTGGAACAACTTA
  • Protein Sequence : Show Sequence 
    >NP_056898.1 p18 [Bovine leukemia virus]
MPKERRSRRRPQPIIRWQVLLVGGPTLYMPARPWFCPMMSPSMPGAPSAGPMNDSNSKGSTPRSPARPTV
SPGPPMDDLLASMEHCSLDCMSPRPAPKGPDDSGSTAPFRPFALSPARFHFPPSSSPPSSPTNANCPRPL
ATVAPSSGTAFFPGTT
  • Molecule Role : Protective antigen
  • Molecule Role Annotation : The bovine leukemia virus (BLV) envelope gene encoding extracellular glycoprotein gp51 (encoded by env gene) and transmembrane glycoprotein gp30 was cloned into a vehicle expression vector under the human cytomegalovirus (CMV) intermediate early promoter. The intramuscular injection of this plasmid vector generated a cellular immune response. Seven out of ten cows vaccinated with the DNA construct resisted a drastic challenge (Brillowska et al., 1999).
  • Related Vaccine(s): Bovine leukemia virus DNA vaccine encoding Env
3. gag
  • Gene Name : gag
  • Sequence Strain (Species/Organism) : Bovine Leukemia virus
  • NCBI Protein GI : AGV74358
  • Other Database IDs : CDD:279002
  • Taxonomy ID : 11901
  • Protein Name : P24
  • Protein pI : 7.44
  • Protein Weight : 23243.54
  • Protein Length : 263
  • Protein Note : gag gene protein p24 (core nucleocapsid protein); pfam00607
  • Protein Sequence : Show Sequence 
    >AGV74358.1 P24 [Bovine leukemia virus]
MGLPIISEGNRNRHRAWALRELQDIKKEIENKAPGSQVWIQTLRLAILQADPTPADLEQLCQYIASPVDQ
TAHMTSLTAAIAAAEAANTLQGFNPQNGTLTQQSAQPNAGDLRSQYQNLWLQAWKNLPTRPSVQPWSTIV
QGPAESYVEFVNRLQISLADNLPDGVPKEPIIDSLSYANANKECQQILQGRGLVAAPVGQKLQACAHWAP
KIKQPAIL
  • Molecule Role : Protective antigen
  • Molecule Role Annotation : (Larsen et al., 2013)
III. Vaccine Information
1. Bovine leukemia virus DNA vaccine encoding Env
a. Vaccine Ontology ID:
VO_0011378
b. Type:
DNA vaccine
c. Status:
Research
d. Antigen
Bovine leukemia virus envelope gene encoding extracellular glycoprotein gp51and transmembrane glycoprotein gp30
e. Gene Engineering of Env/Rex/Tax
  • Type: DNA vaccine construction
  • Description: The bovine leukemia virus (BLV) envelope gene encoding extracellular glycoprotein gp51 (encoded by env gene) and transmembrane glycoprotein gp30 was cloned into a vehicle expression vector under the human cytomegalovirus (CMV) intermediate early promoter (Brillowska et al., 1999).
  • Detailed Gene Information: Click here.
f. Vector:
pcDNA3 (Brillowska et al., 1999)
g. Immunization Route
Intramuscular injection (i.m.)
h. Cattle Response
  • Efficacy: The intramuscular injection of this plasmid vector generated a cellular immune response. Seven out of ten cows vaccinated with the DNA construct resisted a drastic challenge (Brillowska et al., 1999).
2. Bovine Leukemia Virus Vaccine pBLV6073DX
a. Type:
Recombinant vector vaccine
b. Status:
Licensed
c. Host Species for Licensed Use:
Cattle
d. Antigen
Vaccine strain (pBLV6073DX) with a point mutation in the transmembrane protein gene (T>G at nucleotide 6073) and a partial deletion of the R3-G4 sequences (between positions 6614 and 6997). (Suárez et al., 2022)
e. Vector:
pSP64 vector (Promega) (Suárez et al., 2022)
f. Preparation
The proviral construct inserted in the pSP64 vector (Promega) was amplified in Escherichia coli C3040 (New England Biolabs). The standard protocol for production of the inoculum was based on transfection of a monolayer of CHOK1 cells in 25 cm2 cell culture flasks with 2.6 µg of pBLV6073DX and 5.2 µg of linear polyethylenimine (Polysciences) in the presence of 3 ml EMEM (Sigma-Aldrich). Four hours post-transfection, 3 ml of EMEM supplemented with 10% FBS (Internegocios SA) and 3% trehalose were added. Seventy-two hours post-transfection, lysate aliquots (6 ml/dose) were preserved at -80°C in the presence of 15% trehalose. (Suárez et al., 2022)
g. Immunization Route
subcutaneous injection
IV. References
1. Brillowska et al., 1999: Brillowska A, Dabrowski S, Rułka J, Kubiś P, Buzała E, Kur J. Protection of cattle against bovine leukemia virus (BLV) infection could be attained by DNA vaccination. Acta biochimica Polonica. 1999; 46(4); 971-976. [PubMed: 10824867].
2. Kabeya et al., 1996: Kabeya H, Ohashi K, Ohishi K, Sugimoto C, Amanuma H, Onuma M. An effective peptide vaccine to eliminate bovine leukaemia virus (BLV) infected cells in carrier sheep. Vaccine. 1996; 14(12); 1118-1122. [PubMed: 8911007].
3. Larsen et al., 2013: Larsen A, Gonzalez ET, Serena MS, Echeverría MG, Mortola E. Expression of p24 gag protein of bovine leukemia virus in insect cells and its use in immunodetection of the disease. Molecular biotechnology. 2013; 54(2); 475-483. [PubMed: 22829115].
4. Usui et al., 2003: Usui T, Konnai S, Tajima S, Watarai S, Aida Y, Ohashi K, Onuma M. Protective effects of vaccination with bovine leukemia virus (BLV) Tax DNA against BLV infection in sheep. The Journal of veterinary medical science / the Japanese Society of Veterinary Science. 2003; 65(11); 1201-1205. [PubMed: 14665749].
5. Wiki: Bovine leukemia virus: Bovine leukemia virus [http://en.wikipedia.org/wiki/Bovine_leukemia_virus]