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).
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).
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 10^6 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).
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 10^4 and 10^6 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).
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).
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).
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).
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 × 10^5 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).
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 10^6 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).
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).
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 × 10^5 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).
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).
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 10^3, 10^4, 10^5, and 10^6 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 10^4 and 10^6 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. Duarte et al., 2017: Duarte MC, Lage DP, Martins VT, Costa LE, Carvalho AMRS, Ludolf F, Santos TTO, Vale DL, Roatt BM, Menezes-Souza D, Fernandes AP, Tavares CAP, Coelho EAF. A vaccine composed of a hypothetical protein and the eukaryotic initiation factor 5a from Leishmania braziliensis cross-protection against Leishmania amazonensis infection. Immunobiology. 2017; 222(2); 251-260. [PubMed: 27693018].
5. 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].
6. 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].
