Table 2. Table 1 Biomarker of immunogenicity in vaccine kennel trial after L chagasi/L. infantum challenge

Vaccines / Study design / Experimental
challenge / Biological specimens / Immunogecity biomarkers/clinical signs / Tissue parasitism / Refs.
L. braziliensis sound- disrupted promastigotes + BCG / Mongrel dog, 4 month old (n=9); 3 id/21-days interval / 2.3x106
L. chagasi promastigotes iv/60dai / Serum, PBMC, bone marrow / Before challenge: ­a Cell proliferation in vaccinees (8/9) and unresponsiveness in controls (10/10)
After challenge: IFAT positive in vaccinees (1/9) and controls (9/10) / bone marrow culture positive in vaccinees (1/9) and control (9/10) / [60]
Q + BCG / Beagle, 8-9 months old (n=10); 3 ip/21-days interval / 5x105
L. infantum promastigotes iv/66dai / Serum, LN, spleen, liver / After challenge: ­ anti-Leishmania IgG in all vaccinees (10/10) and controls (0/10); DTH positive in vaccinees (8/10) and controls (1/10)
Presence of clinical signs in vaccinees (1/10) and controls (9/10) / LN culture positive in vaccinees (5/10-150dpi, 3/10 - 450dpi, 1/10 - 634 dpi) and controls (10/10-150dpi, 9/10 - 450dpi, 9/10 - 634 dpi)
spleen imprint negative in all vaccines and positive in controls (3/10) / [61]
LiESAp / Beagle,1-6 years old (n=12); 2 sc/21-days interval / 108
L. infantum promastigotes iv/240dai / Serum and CM-DM / Before and after challenge:­ IgG2; ­ LiESAp-specific PBMC proliferation;­ in vitro leishmanicidal effect, ­ NO levels and ­ IFN-g in vaccinees
Absence of clinical signs in vaccines / bone marrow culture positive in LiESAp (1/3 - 50mg, 0/3 - 100mg , 0/3 - 200mg) and controls (3/3) / [53]
H1; HASPB1; HASPB1 + H1; MML / Beagle, 8-36 months old (n=8/vaccine); 3 id/30-days interval / 108
L. infantum promastigotes iv/45dai / Serum, PBMC LN, whole peripheral blood, and bone marrow / Before challenge: - ­ specific IgG in vaccinees
After challenge: ­ specific IgG in vaccinees and controls, ¯b leukocytes in, ¯ lymphocytes , ¯ platelets in vaccinees
Absent Leishmania-specific cell proliferation before and after challenge
Presence of clinical signs in, HASPB1 (4/8), H1(3/8); HASPB1+H1 (4/8), MML (5/7) and in controls (6/8) / bone marrow culture and PCR positive 448 dpi in vaccinees (3/8, 5/8- HASPB1; 3/8, 3/8 - H1; 5/8, 6/8 – HASPB1+H1; 3/7, 4/7 - MML) and in controls (3/8, 2/8)
LN culture and PCR positive 448dpi in vaccinees (6/8, 5/8- HASPB1; 4/8, 5/8 - H1; 3/8, 5/8 – HASPB1+H1; 5/7, 5/7 - MML) and in controls (5/8, 2/8) / [38]
A2 / Beagle, 3-9 months old (n=14); 3sc/21-days interval / 5x107
L. chagasi promastigotes iv/28dai / Serum, whole peripheral blood, PBMC, bone marrow / Before and after challenge: ­anti-A2 IgG and IgG2 in vaccinees
After challenge: ­ anti-L. chagasi IgG and IgG2 in infected vaccinees and controls); ­ IFN-g in infected vaccinees
Presence of clinical signs in infected vaccinees (2/7) and controls (5/7) / bone marrow culture positive in vaccinees (4/7) and controls (7/7)
peripheral blood PCR positive in vaccinees (2/7) and controls (5/7) / [67]

a­ = increase in the biomarker levels when compared to controls groups

b¯ = decrease in the biomarker levels when compared to controls groups

Abbreviations: A2, purified recombinant protein of L. donovani plus saponin; CM-DM, canine monocyte-derived macrophages co-cultured with autologous lymphocytes; dpi, days post infection; DTH, delayed type hypersensitivity; H1, L. infantum recombinant histone H1 plus MontanideTM ISA 720 (used only in first and second immunization); HASPB1, L. donovani hydrophilic acylated surface protein B1 plus MontanideTM ISA 720 (used only in first and second immunization); ID, intradermic route; IFAT, Anti-Leishmania immunofluorescent antibody test; IV, intravenous route; LiESAp vaccine, purified naturally excreted-secreted antigens of L. infantum (MHOM/MA/67/ITMAP-263, clone 2) promastigotes plus muramyl dipeptide (MDP); LN, popliteous lymph node; MML, mixture of TSA, LmSTI1 and LeIF Leishmania antigens plus MPL-SEÒ; NO, nitric oxide evaluated by nitrite/nitrate levels, PBMC, peripheral blood mononuclear cells; Q + BCG, quimeric multi component antigenic protein formed by genetic fusion of fragments from the acid ribosomal proteins Lip2a, Lip2b, P0 and histone H2A protein associated with BCG (Bacillus Calmete Guérin); SC, subcutaneous route.