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Leishmania amazonensis

Table of Contents
  1. General Information
    1. NCBI Taxonomy ID
    2. Disease
    3. Introduction
    4. Host Protective Immunity
  2. Vaccine Related Pathogen Genes
    1. CP cysteine proteinase (Protective antigen)
    2. Gp46 (Protective antigen)
    3. M2 (Protective antigen)
    4. P4 (Protective antigen)
    5. WD (Protective antigen)
  3. Vaccine Information
    1. L. amazonensis CP Protein Vaccine
    2. L. amazonensis DNA vaccine encoding GP46
    3. L. amazonensis DNA vaccine encoding WD protein
    4. L. amazonensis M2 protein vaccine
  4. References
I. General Information
1. NCBI Taxonomy ID:
5659
2. Disease:
Leishmaniasis
3. Introduction
The different Leishmania species cause a broad spectrum of human diseases. L. amazonensis is known to be associated with cutaneous, diffuse cutaneous, and visceral leishmaniasis in South and Central America. The pathological mechanisms responsible for the variable outcomes of infection in humans are not fully understood; however, it is generally agreed that long-lasting immunity against reinfection can be developed in cutaneous leishmaniasis patients. Several vaccination trials have demonstrated that killed L. amazonensis can induce protection from natural infection. However, the efficacy of heat-killed vaccines against Leishmania has been extremely low or highly variable within the same study. Live parasites have been used as a vaccine strategy, and although they are highly effective in inducing immunity, this strategy has been virtually abandoned due to safety issues associated with injecting virulent organisms (Campbell et al., 2003).
4. Host Protective Immunity
Protective mechanisms in L. amazonensis are unclear. During L. amazonensis infection, BALB/c mice display a mixed profile of both Th1 and Th2 responses, although the implications of this mixed profile are ambiguous. However, the necessity for vaccines to induce a Th1-dominant response for these species of Leishmania appears to be consequential for protection (Campbell et al., 2004). The immune responses induced against L. (L.) amazonensis have showed a distinct pattern from that described for L. (L.) major. While mice that are resistant or susceptible to infection with L. (L.) major exhibit immune responses polarized to Th1 and Th2, respectively, susceptibility to L. (L.) amazonensis can not be correlated to an increased Th2 response (Fedeli et al., 2010).
1. CP cysteine proteinase
  • Gene Name : CP cysteine proteinase
  • Sequence Strain (Species/Organism) : Leishmania amazonensis
  • VO ID : VO_0011295
  • NCBI Nucleotide GI : 30142041
  • NCBI Protein GI : 30142042
  • Protein Accession : AAN34826.1
  • Other Database IDs : CDD:140230
    CDD:157580
    CDD:30292
  • Taxonomy ID : 5659
  • Gene Strand (Orientation) : ?
  • Protein Name : cysteine proteinase
  • Protein Length : 354
  • Protein Note : forma: amastigote
  • DNA Sequence : Show Sequence
    >gi|30142041|gb|AY141759.1| Leishmania mexicana amazonensis clone 2A1 cysteine proteinase mRNA, complete cds
    CCGTCACGCCCTTCTCGTGCGCACGTTGCAGAACCAGGATCGGCAAAGATGGCGCGCCGCAACCCCCTTT
    TGTTTGCGATAGTGGTGACGATCCTGTTCGTGGTGTGCTACGGTTCCGCTCTCATCGCCCAGACACCCCC
    CGCCGTCGACAACTTCGTTGCCTCAGCGCATTACGGAAGTTTTAAGAAGCGCCACAGTAAGGCCTTCGGC
    GGGGACGCCGAGGAGGGTCACCGCTTCAATGCCTTCAAGCAGAACATGCAGACAGCCTACTTCCTCAACA
    CGCAGAACCCCCACGCGCACTACGACGTGTCCGGCAAGTTCGCGGACCTCACCCCGCAGGAGTTCGCCAA
    GCTGTACCTGAATCCCGACTACTACACGAGCCACCTCAAGGATCACAAGGAGGACGTGCACGTCGACGAC
    AGCGCCCCCAGTGGTGTGATGTCGGTGGACTGGCGTGATAAGGGTGCCGTGACGCCGGTGAAGAACCAGG
    GATTATGCGGCTCGTGCTGGGCCTTCTCCGCCATTGGCAACATCGAAGGTCAGTGGGCTGCAAGCGGCCA
    CTCGTTGGTTTCACTGTCAGAACAGATGCTCGTGTCGTGCGACAACGTCGATGAAGGGTGCAACGGCGGG
    CTGATGGACCAGGCAATGAACTGGATCATGCAGAGTCACAACGGCAGTGTGTTCACGGAGGCCAGCTATC
    CCTACACCTCTGGCGGCGGCACCAGACCGCCGTGCCATGACGAAGGTGAAGTTGGCGCTAAAATCACCGG
    TTTCCTCTCCCTGCCGCACGACGAGGAGCGGATCGCGGACTGGGTGGAGAAGAGAGGCCCCGTCGCCGTC
    GCCGTCGACGCGACAACCTGGCAGCTGTACTTTGGCGGTGTGGTCTCCCTCTGCCTCGCGTGGTCGCTCA
    ACCACGGTGTGCTCATTGTCGGCTTCAACAAAAACGCGAAACCGCCGTACTGGATCGTGAAGAACTCGTG
    GGGCTCCTCGTGGGGTGAGAAGGGGTACATCCGCCTTGCCATGGGCAGCAACCAGTGCATGCTCAAGAAT
    TACCCCGTGTCGGCCACGGTTGAAAGCCCCCACACCCCACACGTGCCGACGACAACGGCCTAGCTGGAGC
    CCAGGCCCACGGCGCCTGGGCGAATGAGCTGCTTTTTTTTTCGAGTGGGTGCTCGAGGCTGTGCACAAGC
    ACGACCTATCCGACCTGTGTGTGCCTGAAGCGCCAGAATGGCGGCTCTGCCCAGGTGACCTGTGGCAAAC
    CCGAGACGGCGGAGCTTCTCTACAACTGGCCGGGTTGCTCCGAGGCTGCCACCGAGGCCTGCATGCCGCT
    GAACAATATGTATGCGCAGCTACGCCGGCTACCTCCAGAACGTCTGCACTGCGTTCGTCGCTTGCACTGA
    GAACCAACGACGCCGTCTCTGACTTGACCAAGCCAACCATCTTGGGCCAACCACACGTCCCGTACGCGAG
    GCACGCGGGGATCCAGGACTGGCAGGATCCACTGCGCACTATACGCCTCAGCTGCACCGATGCGGATCCC
    TCCCTCAAAAAAAAAAAAAAAAAAAAAA
  • Protein Sequence : Show Sequence
    >gi|30142042|gb|AAN34826.1| cysteine proteinase [Leishmania amazonensis]
    MARRNPLLFAIVVTILFVVCYGSALIAQTPPAVDNFVASAHYGSFKKRHSKAFGGDAEEGHRFNAFKQNM
    QTAYFLNTQNPHAHYDVSGKFADLTPQEFAKLYLNPDYYTSHLKDHKEDVHVDDSAPSGVMSVDWRDKGA
    VTPVKNQGLCGSCWAFSAIGNIEGQWAASGHSLVSLSEQMLVSCDNVDEGCNGGLMDQAMNWIMQSHNGS
    VFTEASYPYTSGGGTRPPCHDEGEVGAKITGFLSLPHDEERIADWVEKRGPVAVAVDATTWQLYFGGVVS
    LCLAWSLNHGVLIVGFNKNAKPPYWIVKNSWGSSWGEKGYIRLAMGSNQCMLKNYPVSATVESPHTPHVP
    TTTA
  • Molecule Role : Protective antigen
  • Molecule Role Annotation : A 500 bp fragment encoding an isoform of cysteine proteinase (CP) from Leishmania (Leishmania) amazonensis was subcloned and expressed in the pHis vector, resulting in a recombinant protein of 24 kDa, rLacys24. Immunization of BALB/c mice with rLacys24 plus CFA adjuvant resulted in a low but significant decrease of foot lesions after challenge with L. (L.) amazonensis compared to those exhibited by control mice (Fedeli et al., 2010).
  • Related Vaccine(s): L. amazonensis CP Protein Vaccine
2. Gp46
  • Gene Name : Gp46
  • Sequence Strain (Species/Organism) : Leishmania amazonensis
  • VO ID : VO_0011292
  • NCBI Nucleotide GI : 159320
  • NCBI Protein GI : 159321
  • Protein Accession : AAA29234.1
  • Other Database IDs : CDD:165684
  • Taxonomy ID : 5659
  • Gene Strand (Orientation) : ?
  • Protein Name : surface membrane glycoprotein GP46/M-2
  • Protein Length : 476
  • Protein Note : leucine-rich repeat receptor-like protein kinase; Provisional; PLN00113
  • DNA Sequence : Show Sequence
    >gi|159320|gb|M38368.1|LEIGP46M2 L.amazonensis surface membrane glycoprotein GP46/M-2 gene, complete cds
    ATGGCGCAGTGCGTGCGTCGGCTGGTGCTGGCGGCGCCCCTCGCCGCTGTGGTGGCGCTGCTGCTGTGCA
    CGAGCAGTGCACCGGTGGCGCGTGCTGCGGGGACGAGCGACTTCACTGGTGCGCAGCAGAAGAACACGCT
    GACGGTGCTGCAGGCGTTTGCGCGTGCGATCCCTGCGCTTGGGGACACGTGGACGGGCAGCGACTTCTGC
    TCGTGGGAGCACATCATCTGCTACTCCTCCGGCGTCGGCGTGTGGATGCACAATGTGGATTATACCGGCA
    CGCTGCCGGAGATGCCTGCGAGCGTCGACTACAAGGACGTCATGATCTTGGCACTGGACTTCGGCGCAAT
    GGGCCAGGGGCTGAGCGGGACGCTGCCCCCCTCATGGAGCTCGATGAAGCACCTGATCGTTCTGGATCTG
    GAGGGCACTAAGGTGTCCGGCACGCTGCCGCCCGAGTGGAGTGAGATGACGAGTGCTGAGGCCCTGCAGC
    TGGAGAACTGCGGTCTGTCCGGGAGTCTGCCCACCTCGTGGTCTTCGATGCCGAAGCTGCGTATCGTCTC
    ACTGAGCGGCAACCACTTCTGCGGGTGCGTGCCCGACTCGTGGAGGGAGAAGGACCGCCTCGATGTGACC
    ATCGAGGAATGGCACATGGGCGAGGACTGCAAGCTTGCTAACGCCTGCCGCCCGACTGCTGCTCCGGGAA
    CGACCACGACTAACCCGCCCACCACCACCGGCACCCCAGCAGCCTCCTCTACTCCTTCTCCAGGGTCGGG
    GTGCGAGGTGGATGGGTGTGAGGTGTGCGAGGGGGACTCCGCTGCGCGGTGCGCCAGGTGCCGTGAGGGC
    TACTCCCTGACGGACGAGAAGACGTGCCTGGGCGAACCACGATGGCGGCGTGGCGGCGGCGTCGAGCGGA
    CGGCTGGCTGCCGCTGCTGTGTGGGCGGCTGTGCTGTTGAGCGTGGGGCTGGTGGCGTGAGGGTGCGGCG
    GGCCCCTCTTCTCTGTGGTGCCCCTGGTGCCTGCCCTCGCCCCCGGCACGGCGTCGTCGCTGCCCTCTCA
    CCCCCACCAGCCGACGGGGAGACCGACAGCCACACGCGCACGCGCACACGCCGTCGTGCATCGCGTGTGC
    TTTCCGCCGTTGTGGCGCCTGCGCGGATGCACGGGCATGCGGAGGCGTGCATGCGTGTGCGCGTGCCGGC
    TCTTGTGTGTCTCTCCGTGTGGCCAGCAGTCGGCACCCGCCGCCGATCGAATGTGCGCGCGGCGGCGGTG
    TGTCGCCTTGGACAGCGGAGATGCGGCGCCCGCCCCTCGCCGTGTGCCTCGGTCTGCGTGTCGTGGCCGC
    GCGAGCGACGTACGGAGTGCGCGTGTCCGGCCCTCTTCGACGGGGCTCGCTTGCGGTGCTGTGCTCTCGT
    GGTCTGTGCCGGTGCTGCCCCGGCGGGGTGA
  • Protein Sequence : Show Sequence
    >gi|159321|gb|AAA29234.1| surface membrane glycoprotein GP46/M-2 [Leishmania amazonensis]
    MAQCVRRLVLAAPLAAVVALLLCTSSAPVARAAGTSDFTGAQQKNTLTVLQAFARAIPALGDTWTGSDFC
    SWEHIICYSSGVGVWMHNVDYTGTLPEMPASVDYKDVMILALDFGAMGQGLSGTLPPSWSSMKHLIVLDL
    EGTKVSGTLPPEWSEMTSAEALQLENCGLSGSLPTSWSSMPKLRIVSLSGNHFCGCVPDSWREKDRLDVT
    IEEWHMGEDCKLANACRPTAAPGTTTTNPPTTTGTPAASSTPSPGSGCEVDGCEVCEGDSAARCARCREG
    YSLTDEKTCLGEPRWRRGGGVERTAGCRCCVGGCAVERGAGGVRVRRAPLLCGAPGACPRPRHGVVAALS
    PPPADGETDSHTRTRTRRRASRVLSAVVAPARMHGHAEACMRVRVPALVCLSVWPAVGTRRRSNVRAAAV
    CRLGQRRCGARPSPCASVCVSWPRERRTECACPALFDGARLRCCALVVCAGAAPAG
  • Molecule Role : Protective antigen
  • Molecule Role Annotation : cDNAs encoding L. m. amazonensis gp46 antigen were subcloned into the VR1012 plasmid, and susceptible BALB/c mice were immunised with two i.m. injections of 100 microg of plasmid DNA. Mice were challenged by the injection of 4 x 106 L. m. mexicana parasites in the foot pad to evaluate protection. Measurement of lesion size indicated that mice immunised with VR012-GP46 were partially protected against infection (Dumonteil et al., 2000).
  • Related Vaccine(s): L. amazonensis DNA vaccine encoding GP46
3. M2
  • Gene Name : M2
  • Sequence Strain (Species/Organism) : Leishmania amazonensis
  • VO ID : VO_0011291
  • NCBI Nucleotide GI : 2828562
  • NCBI Protein GI : 2828563
  • Protein Accession : AAC08302.1
  • Other Database IDs : CDD:153108
  • Taxonomy ID : 5659
  • Gene Strand (Orientation) : ?
  • Protein Name : ribonucleotide reductase M2 subunit
  • Protein Length : 345
  • Protein Note : hydroxyurea-resistant strain derived from LV-78 cloned from extrachromosomal amplified circular DNA
  • DNA Sequence : Show Sequence
    >gi|2828562|gb|AF005247.1| Leishmania mexicana amazonensis extrachromosomal element ribonucleotide reductase M2 subunit gene, complete cds
    ATGAAACCGGTTGCGGCTGGCGCCGAGGTGCTGCCGGCGGACAAGGTTGCCCAGGGGACGAACGCCGAGG
    AGGAGCCGCTGCAGCAGGAGAACCCGTTCCGCTACGTCCTCTTCCCAATCCAGTACCATGACATCTGGCG
    GAAGTACAAGGAGCAGGAGAGCTGCATCTGGACGGTGGAGGAGATCGACCTGGGCAACGACATGAAGGAC
    TGGGCAACGCTGAACGACGGTGAGCGGCACTTCATCAAGCATGTGCTTGCCTTCTTCGCCGGCAGCGACG
    GCATAGTCATCGAGAATCTTGCGCAGCGCTTCATGAGCGACGTGAAGGTGCCAGAGGCGCGCGCGTTCTA
    CGGGTTCCAGCTGATGATGGAGAACATCCACTCGGAGACGTACTCTGTGCTGCTGGACACGTACATCACG
    GACAGCGAGGAGAAGCTGCGGCTGCTGCACGCGATACAGACGATCCCGTGCATCCAGAAGAAGGCGGAGT
    GGCCCGTGCGGTGGATCGGGAGCAGCGCGAGCTTTCAGCAGCGGCTGATCGGCTTTGCCGCGGTGGAGGG
    CATTTTCTTTTCCGGGTCGTTCTGCGCGCTGTTCTGGCTGAAGAAGCGCGGTCTGATGCCAGGGCTGACG
    TTCAGCAACGAGCTCATCTCGCGCGACGAGGGTCTACACACGGACTTCGCATGCCTGCTGTACAACTCGC
    ACATCAAGCACAAGCTGCCGCGCGAGCGCGTGCTGGAGATCATCGTGGATGCGGTGAACATCGAGCGTGA
    GTTCATCTGCGACGCGCTGCCGGTGCGGCTGATTGGCATGAACGCGGAGCTGATGGCGCAGTACATCGAG
    TTCGTCGCGGACCGGCTGCTGGTGTCGCTCGGCGAGGAGAAGCACTACCGCTCGACGCAGCCGTTCGACT
    TCATGGAGATGATCTCGCTGCAGGGCAAAACGAACTTCTTCGAGAAGAGGGTGGGGGAGTACCAGAAGGC
    CGGCGTGATGAGCACGGAGGGCACGTCGAAAAAGTTCTCCCTCTCGGAGGACTTCTAA
  • Protein Sequence : Show Sequence
    >gi|2828563|gb|AAC08302.1| ribonucleotide reductase M2 subunit [Leishmania amazonensis]
    MKPVAAGAEVLPADKVAQGTNAEEEPLQQENPFRYVLFPIQYHDIWRKYKEQESCIWTVEEIDLGNDMKD
    WATLNDGERHFIKHVLAFFAGSDGIVIENLAQRFMSDVKVPEARAFYGFQLMMENIHSETYSVLLDTYIT
    DSEEKLRLLHAIQTIPCIQKKAEWPVRWIGSSASFQQRLIGFAAVEGIFFSGSFCALFWLKKRGLMPGLT
    FSNELISRDEGLHTDFACLLYNSHIKHKLPRERVLEIIVDAVNIEREFICDALPVRLIGMNAELMAQYIE
    FVADRLLVSLGEEKHYRSTQPFDFMEMISLQGKTNFFEKRVGEYQKAGVMSTEGTSKKFSLSEDF
  • Molecule Role : Protective antigen
  • Molecule Role Annotation : Immunization of CBA mice with the M-2 glycoprotein of L. amazonensis and C. parvum adjuvant resulted in complete protection against a challenge infection of 104 and 106 late log-phase promastigotes of L. amazonensis. In the BALB/c strain, complete protection was observed in some of the immunized animals (28 to 50%); in the rest of the mice the onset of infection was significantly delayed. Protective immunity for C57BL/6 mice was observed only at the low infecting dose (10(4) L. amazonensis organisms) (Champsi and McMahon-Pratt, 1988).
  • Related Vaccine(s): L. amazonensis M2 protein vaccine
4. P4
  • Gene Name : P4
  • Sequence Strain (Species/Organism) : Leishmania amazonensis
  • VO ID : VO_0011288
  • NCBI Nucleotide GI : 29165286
  • NCBI Protein GI : 29165287
  • Protein Accession : AAO65599.1
  • Other Database IDs : CDD:145426
  • Taxonomy ID : 5659
  • Gene Strand (Orientation) : ?
  • Protein Name : P4 nuclease
  • Protein Length : 316
  • Protein Note : S1/P1 Nuclease; pfam02265
  • DNA Sequence : Show Sequence
    >gi|29165286|gb|AY219923.1| Leishmania amazonensis P4 nuclease mRNA, complete cds
    ATGCCTGCGCCTGTCAGTCTCCGTCTGCCTCTCACAGCGCTGTGCCTCCTGGTGCTCAGCTCTGCGCTGT
    GCGTGACTGAGGTGCTCGGATGGGGCTGCGTGGGCCACATGCTCCTCGCCGAGATCGCGCGGCGCCAGCT
    GGACTTAGAGAATGAGGAAAAGATCGAGCTGATGGCGGCAGTATTTTCGGGCAGCGGTCCGTTCCCGATG
    TCTCCGAGCATGGTGCAAGCGGCATGCTGGGCGGATGACGTGAAGCTTTGGCGTCAGTACGCCATGAGCA
    CGTGGCACTTCTACGCCATGCCATATAACCCCGGAAACATCAATATCACCGATCCGGTCAATACAGTGAA
    CGCCGTCACTGTGTGCCTTGATATGGTGACCTCGCTTAAGAACTCGAAAGCGCCGCTGTACTTGTTGAAC
    TTTGCTTGGGTGAACCTGGTGCACATCTTTGGCGATCTGCATCAGCCCCTGCACACGATATCTAGGTACA
    CTACCGCCTACCCGCACGGAGATCAAGGAGGCAACGCGATTTCGGTTCGTGTAGGCGGAAAAAAGGTGAA
    GCTGCATGCCCTCTGGGATAATATTTGCAGTGCTACGCCACCACGCTACCAGCGGCCCCTCTCCCACACC
    GACCTCTTCGCCCTCTCTGCGACAGCGGACGGGCTGGTGGAAACTTACACGTTTTCAGAGGCGTTGGAGA
    CACTCGTGGACGTGATGGCGATTCATGAGGAGAGCTACATGTTTGCGGTAAACACCAGCTACCCCGGCGT
    CACACCAGGTGGGACTCTCAGTCGTGCCTACCTTGATAAATGCAAGCGTGTCGCTGAGGCTCGACTGACG
    CTTGGCGGCTACCGCCTCGGTTACCTTCTGAATCAGCTGCTGTCGGGCATCACTGTGGACAAGGCCGCAC
    TGGAGGCGCACCGCGCTGCGCGTCCGAAGTGGAGCGCGTAA
  • Protein Sequence : Show Sequence
    >gi|29165287|gb|AAO65599.1| P4 nuclease [Leishmania amazonensis]
    MPAPVSLRLPLTALCLLVLSSALCVTEVLGWGCVGHMLLAEIARRQLDLENEEKIELMAAVFSGSGPFPM
    SPSMVQAACWADDVKLWRQYAMSTWHFYAMPYNPGNINITDPVNTVNAVTVCLDMVTSLKNSKAPLYLLN
    FAWVNLVHIFGDLHQPLHTISRYTTAYPHGDQGGNAISVRVGGKKVKLHALWDNICSATPPRYQRPLSHT
    DLFALSATADGLVETYTFSEALETLVDVMAIHEESYMFAVNTSYPGVTPGGTLSRAYLDKCKRVAEARLT
    LGGYRLGYLLNQLLSGITVDKAALEAHRAARPKWSA
  • Molecule Role : Protective antigen
  • Molecule Role Annotation : Susceptible BALB/c mice were immunized with the DNA encoding P4 alone, P4/IL-12, or P4/HSP70 prior to challenge with L. amazonensis promastigotes. Mice immunized with P4 alone had a delayed onset of lesion development and significantly smaller lesions than the infection control group at 11 weeks postinfection (P < 0.05). P4/IL-12 immunized animals showed no sign of lesion development in three independent experiments (P < 0.01). Our results indicate that different vaccine combinations, including DNA encoding P4, HSP70, or IL-12, can provide significant protection against both Old World and New World cutaneous leishmaniasis (Campbell et al., 2003).
5. WD
  • Gene Name : WD
  • Sequence Strain (Species/Organism) : Leishmania amazonensis
  • VO ID : VO_0011290
  • NCBI Nucleotide GI : 38455440
  • NCBI Protein GI : 38455441
  • Protein Accession : AAR20840.1
  • Other Database IDs : CDD:29257
  • Taxonomy ID : 5659
  • Gene Strand (Orientation) : ?
  • Protein Name : antigenic WD protein
  • Protein Length : 674
  • Protein Note : WD40 domain, found in a number of eukaryotic proteins that cover a wide variety of functions including adaptor/regulatory modules in signal transduction, pre-mRNA processing and cytoskeleton assembly; typically contains a GH dipeptide 11-24 residues...; cd00200
  • DNA Sequence : Show Sequence
    >gi|38455440|gb|AY457171.1| Leishmania amazonensis antigenic WD protein mRNA, complete cds
    CCCTCCCACTACCACGCAGCTTGTCATATCATACTGTGTTTCATTGTATGCATCCGGCGTGAGCAATTCT
    TTGTGTCCTCAACCGCTCTTTCCTTTCGTCTTTCATCGCTGTGCTTCCTCTCCCCTCAGCGCCCCCCCCT
    CTCCCTTCTCGCACACGCACGCACAGTGGCACAGATACATACAGAGGCACTCCTCTTCTCCCTACACTAT
    CTTGCAGTGCCCTCTTCTTCCCCCGCCTCTTACTCTGCCCGTCCCTCTCCGACTCTCTTTCACGTTTCGC
    AACACCAGTATCAACCATGGCCAAGGAGAAACTCGGCTTCAAGATCCAGGGGAAGACCGTCTCCTACGAG
    GGTTACCCCGAGCGTCCGCCAAGGGACCTGATGGACATCTTGGTCAAAATGGGTGTCGGCGAAGCTGCCG
    TGGTGACCACCTTTCTAGAAAATATTGAGAACCACCGGGAGTTCACCGACGAGCAGGTGATGGTGATGTA
    CAACGAAGCAAAAATGACGCGCGAGAAGAAGGCTCGTGCGAAGGCCGGCAACCAGGGTGACTCAGCCGCT
    GCCGCCGCCGCTCCATCGGGCGCCGCCGATGAGGCGGAGAAGCCGCAGAAGATCCTTCTTTCCTACCGCG
    GCAAGCAGCTTTCCTACGTAGGTTTGCCCAGCGGCAAGCGCAACGCCGTGCTGCAACTCCTCTCCAAGCA
    AGAGGAGCTCACCCGGGAGCTTGCAGTGGAGACGTTCCGCACGAACCTGCCCAGTGATCCGTCCACGGAG
    GAGGAGATTTGGATCTTGGTGCAGGCAGTCAAAGATCGCAAGAGTAGGAAGAGCCAAGCGAAGGGTCAGG
    ATGGCGAAAGGTCCGGCGGTAACGGTGCGACTGCGTCGCTTGGGGCAGGCAGCAGCGCCATGGACAGCAT
    AACCGGTATGTCCGAGAGGGAGCGCGACGACCTGAACAACTTCATTCTTCAGGTGCTGCAGCAGCCAGCG
    CCGGATGTGGCCGCCCTCTTACATGAGAATCCGAAACCGCGCAAAGACACGGAAGACAATGCGGAGCTGG
    CACTGACAGCGCGCATGTCGGGTAAGATTCCGGTTTACCTGCAGGAAGAGAACACCAGCAGCACGAAGCT
    CATCTACGCCACGCCCCACATGAGCTTCGAAGATTTTTCTACCATTGTCGAGAAGAAGTGTGGCCGCAAG
    TTGTCGCTGTCCTTCTACGACGGCGACGACCTCATCATGATGGATGATGACGACGTGCTGGCCATGTTTC
    TCGAAATGACAGTGCAGAACGACCCAAAGAAGGTGCGCCTCATTTGCTCCAACCCTGGAGTCCCAAGGAA
    GGCGGCGGATGACCGGATGATGGAAAACCGTCAAAATACCTTCAGCGGTCTTCCGCAGGTCTCAACGACC
    AAGGCATTGGCGTACTCGAACGGCGAGCTCAACGTCGCGGAGGCTCGCACCTACTTTGGCCACTCTCTTG
    CCGTCTACTGCTGCTGCTTCTCGCCAAGGGGCGACATGTTCGTTACGGCAAGTCGCGACCGCACCGTCCG
    CCTGTGGAACTTGCGCACCGGTGTCTCCACAGTCATGAAAGGCGGGCACAACGGCTTTGTGCTGTCTTGC
    GACTACTCGCCAAAGGGCAACCGCGTCGCTTCCTCTTCTGATGACCGCACTATCAAACTGTGGAACACGT
    CGTCATGCAACAAGGTTGCCACACTTAAAGGGCACGAGGACAAGGTATACTGCGTCAAGTACAACTCTAG
    CGGAGATCTCCTCGTGTCCGCCTCTTGCGACACCACAGTGCGCGTGTGGAACGCCGAAAGTCAGGCGAAG
    CTGGTGACGCTGAGGGGCCACACCCTCGCCGTTTTTTCCTGCGCCTTCAGCAACAGCGATAATGGAAAGT
    TCGTCGTCTCCGGCAGCGATGACCGCGTTATAAAGCTGTGGGACTGGGGCGCCGGCAGAGAGATCCTGTC
    GCTGGTGGGTCACATCGGCACCGTGTGGACGGTTGTCTTCTCCCACAACGACAGGTACGTTCTCAGTGGC
    AGCATGGACTATGAGCTCATTCTCTGGGATTCAATGACTGGCGCTCGCCTGCGCTCGATGGACGGGCACA
    AGACATCGGTGCACCACGCCATCTTCTCCGAGGACGACGCATACATCTTCTCCTGCGCCCGCGACTGGAG
    CGTCATGGTGTGGCGCACCGAAGATGGCGAGCATGTAGAGACCATCATCGGTCACCTCAGCACTGTCTAC
    CACATGGACATCAGCGAGGACAAGTTGCTCACCTGCTCTCTTGATGACAAGATCAAGCTGTGGAATATCA
    GGCGCCACTGAAGTTGAGCACCAGCGGAAAATGTGGTAGTGCGAACACTGGCGGGAGGGGCACGTGCCAG
    CCGCAACCGAGAAAGAGGTGTGGGCGGGAAGGCACGTGATAGGTGATACAGACACTGTACCCGTACACCA
    TTCCTCTTTTCAGCAGCAGTCCTGCACACAGAAAGACACTCGGATTTTTCATTGACAAATCAGGTGGGTC
    TGAAGACGATTTCAAGAAGAAGCGAGCGTTCGTATGTACCGCCCTCCTCCCTCCACCGATACTGCAACAC
    ACACACACACACATTTTACGGTCTTGTCTACTTTTCTTCGTTTCGCTCTCGTCAGTTCCACCGAGTCCTG
    GCTGCGTTTGGGTGCCACGCGTGTGAATGTGTGAGCGCCACCGTTGGTCGACCCTTTTCGCTTTCCTGCA
    TGCTTTTTTTCTTTTCGTATCTCTACGTCCTGCTCACAGCGCATCGACCTCTGCCAGAGGATATCATTTG
    GCAGTGTCGTTTCTTTTGCTCTTTTTCTCTCTTTGCCACCGTTCTTGCGGGCCAACGGCACCGTGCTCCC
    CGCTTCTCTGTCTCGCGACCTCGACATTCATCTCTTCCATGCTTCTCCCTCTCCCCCTCCGTCTCCCCAG
    CTTTTTGTCGTTTGTGCATTCTCCTCTTTTCTTAAGTGATGCTTCAAAGGAAACGAAAAAAAAAAAAAAA
    AA
  • Protein Sequence : Show Sequence
    >gi|38455441|gb|AAR20840.1| antigenic WD protein [Leishmania amazonensis]
    MAKEKLGFKIQGKTVSYEGYPERPPRDLMDILVKMGVGEAAVVTTFLENIENHREFTDEQVMVMYNEAKM
    TREKKARAKAGNQGDSAAAAAAPSGAADEAEKPQKILLSYRGKQLSYVGLPSGKRNAVLQLLSKQEELTR
    ELAVETFRTNLPSDPSTEEEIWILVQAVKDRKSRKSQAKGQDGERSGGNGATASLGAGSSAMDSITGMSE
    RERDDLNNFILQVLQQPAPDVAALLHENPKPRKDTEDNAELALTARMSGKIPVYLQEENTSSTKLIYATP
    HMSFEDFSTIVEKKCGRKLSLSFYDGDDLIMMDDDDVLAMFLEMTVQNDPKKVRLICSNPGVPRKAADDR
    MMENRQNTFSGLPQVSTTKALAYSNGELNVAEARTYFGHSLAVYCCCFSPRGDMFVTASRDRTVRLWNLR
    TGVSTVMKGGHNGFVLSCDYSPKGNRVASSSDDRTIKLWNTSSCNKVATLKGHEDKVYCVKYNSSGDLLV
    SASCDTTVRVWNAESQAKLVTLRGHTLAVFSCAFSNSDNGKFVVSGSDDRVIKLWDWGAGREILSLVGHI
    GTVWTVVFSHNDRYVLSGSMDYELILWDSMTGARLRSMDGHKTSVHHAIFSEDDAYIFSCARDWSVMVWR
    TEDGEHVETIIGHLSTVYHMDISEDKLLTCSLDDKIKLWNIRRH
  • Molecule Role : Protective antigen
  • Molecule Role Annotation : DNA vaccination in BALB/c mice with the LAWD (for Leishmania antigenic WD protein) and IL-12 genes significantly delayed lesion development from challenge with Leishmania amazonensis, which correlated with a dramatic reduction in parasite burdens (Campbell et al., 2004).
  • Related Vaccine(s): L. amazonensis DNA vaccine encoding WD protein
III. Vaccine Information
1. L. amazonensis CP Protein Vaccine
a. Vaccine Ontology ID:
VO_0004199
b. Type:
Subunit vaccine
c. Status:
Research
d. Antigen
A 500 bp fragment encoding an isoform of cysteine proteinase (CP) from Leishmania (Leishmania) amazonensis was subcloned and expressed in the pHis vector, resulting in a recombinant protein of 24 kDa, rLacys24 (Fedeli et al., 2010).
e. Gene Engineering of CP cysteine proteinase
  • Type: Recombinant protein preparation
  • Description:
  • Detailed Gene Information: Click here.
f. Adjuvant: complete Freunds adjuvant
g. Immunization Route
subcutaneous injection
h. Mouse Response
  • Host Strain: BALB/c
  • Vaccination Protocol: Female BALB/c mice (six per group) were immunized thrice with a 2 week interval with 25 μg of rLacys24 plus complete Freund’s adjuvant (CFA) by the subcutaneous route in the base of the tail. Control animals received PBS or only adjuvant (Fedeli et al., 2010).
  • Challenge Protocol: Two weeks after the last dose animals were challenged with 2.5 × 105 L. (L.) amazonensis amastigotes in PBS in the hind footpad (Fedeli et al., 2010).
  • Efficacy: Immunization of BALB/c mice with rLacys24 plus CFA adjuvant resulted in a low but significant decrease of foot lesions after challenge with L. (L.) amazonensis compared to those exhibited by control mice (Fedeli et al., 2010).
2. L. amazonensis DNA vaccine encoding GP46
a. Vaccine Ontology ID:
VO_0011354
b. Type:
DNA vaccine
c. Status:
Research
d. Antigen
L. amazonensis gp46
e. Gene Engineering of Gp46
  • Type: DNA vaccine construction
  • Description: cDNAs encoding L. m. amazonensis gp46 antigen was subcloned into the VR1012 plasmid (Dumonteil et al., 2000)
  • Detailed Gene Information: Click here.
f. Vector:
VR1012 plasmid
g. Immunization Route
Intramuscular injection (i.m.)
h. Mouse Response
  • Host Strain: BALB/c
  • Vaccination Protocol: Susceptible BALB/c mice were immunised with two i.m. injections of 100 microg of plasmid DNA (Dumonteil et al., 2000).
  • Challenge Protocol: Mice were challenged by the injection of 4 x 106 L. m. mexicana parasites in the foot pad to evaluate protection (Dumonteil et al., 2000).
  • Efficacy: Measurement of lesion size indicated that mice immunised with VR012-GP46 were partially protected against infection (Dumonteil et al., 2000).
3. L. amazonensis DNA vaccine encoding WD protein
a. Vaccine Ontology ID:
VO_0011349
b. Type:
DNA vaccine
c. Status:
Research
d. Antigen
L. amazonensis WD
e. Gene Engineering of WD
  • Type: DNA vaccine construction
  • Description: The whole gene insert from the pBK-CMV clone containing the LAWD gene was subcloned into the pET32a plasmid (Invitrogen) for His tag protein expression. To ensure that the ORF would be read in frame, the plasmid was first digested with BamHI and then blunt-ended by a Klenow reaction (Invitrogen) (Campbell et al., 2004).
  • Detailed Gene Information: Click here.
f. Vector:
pcDNA3.1/Hygro(−) (Invitrogen)
g. Immunization Route
Subcutaneous injection
h. Mouse Response
  • Host Strain: BALB/c
  • Vaccination Protocol: Mice (five per group) were immunized in five locations with a total of 100 μg of DNA (50 μg of LAWD and 50 μg of IL-12) per mouse: four injections in both sides of the inner and outer thigh muscles of the hind legs (∼50 μl/site) and one subcutaneous injection in the left hind foot (∼5 μl/site) (Campbell et al., 2004).
  • Challenge Protocol: Mice were boosted twice at 3-week intervals and then challenged 3 weeks after the last immunization with 2 × 105 metacyclic promastigotes in the right hind foot (Campbell et al., 2004).
  • Efficacy: DNA vaccination in BALB/c mice with the LAWD (for Leishmania antigenic WD protein) and IL-12 genes significantly delayed lesion development from challenge with Leishmania amazonensis, which correlated with a dramatic reduction in parasite burdens (Campbell et al., 2004).
4. L. amazonensis M2 protein vaccine
a. Vaccine Ontology ID:
VO_0011355
b. Type:
Subunit vaccine
c. Status:
Research
d. Antigen
L. amazonensis M2
e. Gene Engineering of M2
  • Type: Recombinant protein preparation
  • Description: M-2, a 46-kDa promastigote-specific glycoprotein was isolated. The protein was further purified by removal of detergent with an anionexchange column(Champsi and McMahon-Pratt, 1988).
  • Detailed Gene Information: Click here.
f. Adjuvant: killed Corynebacterium parvum vaccine adjuvant
g. Immunization Route
Intraperitoneal injection (i.p.)
h. Mouse Response
  • Host Strain: CBA
  • Vaccination Protocol: BALB/c, CBA, and C57BL/6 mice were immunized intraperitoneally with M-2 at a final concentration of 0.03 mg/ml. The amount of C. parvum used in immunizations 2 and 3 was reduced to 0.05 mg per immunization (Champsi and McMahon-Pratt, 1988).
  • Challenge Protocol: Animals were rested for 2 to 4 weeks after final immunization and challenged in the right hindfoot with late-log-phase promastigotes. Parasites used for infections were passaged a maximum of four times. Challenge doses of 103, 104, 105, and 106 were used (Champsi and McMahon-Pratt, 1988).
  • Efficacy: Immunization of CBA mice with the M-2 glycoprotein of L. amazonensis and C. parvum adjuvant resulted in complete protection against a challenge infection of 104 and 106 late log-phase promastigotes of L. amazonensis. In the BALB/c strain, complete protection was observed in some of the immunized animals (28 to 50%); in the rest of the mice the onset of infection was significantly delayed. Protective immunity for C57BL/6 mice was observed only at the low infecting dose (10(4) L. amazonensis organisms) (Champsi and McMahon-Pratt, 1988).
IV. References
1. Campbell et al., 2003: Campbell K, Diao H, Ji J, Soong L. DNA immunization with the gene encoding P4 nuclease of Leishmania amazonensis protects mice against cutaneous Leishmaniasis. Infection and immunity. 2003; 71(11); 6270-6278. [PubMed: 14573646].
2. Campbell et al., 2004: Campbell K, Popov V, Soong L. Identification and molecular characterization of a gene encoding a protective Leishmania amazonensis Trp-Asp (WD) protein. Infection and immunity. 2004; 72(4); 2194-2202. [PubMed: 15039343].
3. Champsi and McMahon-Pratt, 1988: Champsi J, McMahon-Pratt D. Membrane glycoprotein M-2 protects against Leishmania amazonensis infection. Infection and immunity. 1988; 56(12); 3272-3279. [PubMed: 3182080].
4. Dumonteil et al., 2000: Dumonteil E, Andrade-Narvarez F, Escobedo-Ortegon J, Ramirez-Sierra MJ, Valencia-Pacheco G, Flores-Serrano A, Canto-Lara S, Arjona-Torres A. Comparative study of DNA vaccines encoding various antigens against Leishmania mexicana. Developments in biologicals. 2000; 104; 135-141. [PubMed: 11713811].
5. Fedeli et al., 2010: Fedeli CE, Ferreira JH, Mussalem JS, Longo-Maugéri IM, Gentil LG, dos Santos MR, Katz S, Barbiéri CL. Partial protective responses induced by a recombinant cysteine proteinase from Leishmania (Leishmania) amazonensis in a murine model of cutaneous leishmaniasis. Experimental parasitology. 2010; 124(2); 153-158. [PubMed: 19735658].