Skip to main content Accessibility help
×
Home

Antigenicity, immunogenicity and protective efficacy of a conserved Leishmania hypothetical protein against visceral leishmaniasis

  • Daniel S. Dias (a1), Vívian T. Martins (a1), Patrícia A. F. Ribeiro (a1), Fernanda F. Ramos (a1), Daniela P. Lage (a1), Grasiele S. V. Tavares (a1), Débora V. C. Mendonça (a1), Miguel A. Chávez-Fumagalli (a1), Jamil S. Oliveira (a2), Eduardo S. Silva (a3), Dawidson A. Gomes (a2), Michele A. Rodrigues (a4), Mariana C. Duarte (a1) (a5), Alexsandro S. Galdino (a3), Daniel Menezes-Souza (a1) (a5) and Eduardo A. F. Coelho (a1) (a5)...

Abstract

In this study, a Leishmania hypothetical protein, LiHyS, was evaluated regarding its antigenicity, immunogenicity and protective efficacy against visceral leishmaniasis (VL). Regarding antigenicity, immunoblottings and an enzyme-linked immunosorbent assay using human and canine sera showed high sensitivity and specificity values for the recombinant protein (rLiHyS) in the diagnosis of VL. When evaluating the immunogenicity of LiHyS, which is possibly located in the parasite's flagellar pocket, proliferative assays using peripheral blood mononuclear cells from healthy subjects or VL patients showed a high proliferative index in both individuals, when compared to the results obtained using rA2 or unstimulated cultures. Later, rLiHyS/saponin was inoculated in BALB/c mice, which were then challenged with Leishmania infantum promastigotes. The vaccine induced an interferon-γ, interleukin (IL)-12 and granulocyte-macrophage colony-stimulating factor production, which was maintained after infection and which was associated with high nitrite and IgG2a antibody levels, as well as low IL-4 and IL-10 production. Significant reductions in the parasite load in liver, spleen, bone marrow and draining lymph nodes were found in these animals. In this context, the present study shows that the rLiHyS has the capacity to be evaluated as a diagnostic marker or vaccine candidate against VL.

Copyright

Corresponding author

Author for correspondence: Professor Dr Eduardo A. F. Coelho, E-mail: eduardoferrazcoelho@yahoo.com.br

References

Hide All
Abanadés, DR, Arruda, LV, Arruda, ES, Pinto, JR, Palma, MS, Aquino, D, Caldas, AJ, Soto, M, Barral, A and Barral-Netto, M (2012) Immunodominant antigens of Leishmania chagasi associated with protection against human visceral leishmaniasis. PLoS Neglected Tropical Diseases 6, e1687.
Achour, YB, Chenik, M, Louzir, H and Dellagi, K (2002) Identification of a disulfide isomerase protein of Leishmania major as a putative virulence factor. Infection and Immunity 70, 35763585.
Agallou, M, Smirlis, D, Soteriadou, KP and Karagouni, E (2012) Vaccination with Leishmania histone H1-pulsed dendritic cells confers protection in murine visceral leishmaniasis. Vaccine 30, 50865093.
Agallou, M, Athanasiou, E, Samiotaki, M, Panayotou, G and Karagouni, E (2016) Identification of immunoreactive Leishmania infantum protein antigens to asymptomatic dog sera through combined immunoproteomics and bioinformatics analysis. PLoS ONE 11, e0149894.
Agallou, M, Margaroni, M, Athanasiou, E, Toubanaki, DK, Kontonikola, K, Karidi, K, Kammona, O, Kiparissides, C and Karagouni, E (2017) Identification of BALB/c immune markers correlated with a partial protection to Leishmania infantum after vaccination with a rationally designed multi-epitope cysteine protease a peptide-based nanovaccine. PLoS Neglected Tropical Diseases 11, e0005311.
Alcolea, PJ, Tuñón, GI, Alonso, A, García-Tabares, F, Ciordia, S, Mena, MC, Campos, RN, Almeida, RP and Larraga, V (2016) Differential protein abundance in promastigotes of nitric oxide-sensitive and resistant Leishmania chagasi strains. Proteomics Clinical Applications 10, 11321146.
Aronson, NE (2017) Addressing a clinical challenge: guidelines for the diagnosis and treatment of leishmaniasis. BMC Medicine 15, 76.
Bayih, AG, Daifalla, NS and Gedamu, L (2014) DNA-protein immunization using Leishmania peroxidoxin-1 induces a strong CD4+ T cell response and partially protects mice from cutaneous leishmaniasis: role of fusion murine granulocyte-macrophage colony-stimulating factor DNA adjuvant. PLoS Neglected Tropical Diseases 8, e3391.
Bhowmick, S, Ravindran, R and Ali, N (2007) Leishmanial antigens in liposomes promote protective immunity and provide immunotherapy against visceral leishmaniasis via polarized Th1 response. Vaccine 25, 65446556.
Chamakh-Ayari, R, Bras-Gonçalves, R, Bahi-Jaber, N, Petitdidier, E, Markikou-Ouni, W, Aoun, K, Moreno, J, Carrillo, E, Salotra, P, Kaushal, H, Negi, NS, Arevalo, J, Falconi-Agapito, F, Privat, A, Cruz, M, Pagniez, J, Papierok, GM, Rhouma, FB, Torres, P, Lemesre, JL, Chenik, M and Meddeb-Garnaoui, A (2014) In vitro evaluation of a soluble Leishmania promastigote surface antigen as a potential vaccine candidate against human leishmaniasis. PLoS ONE 9, e92708.
Coelho, VT, Oliveira, JS, Valadares, DG, Chávez-Fumagalli, MA, Duarte, MC, Lage, PS, Soto, M, Santoro, MM, Tavares, CA, Fernandes, AP and Coelho, EA (2012) Identification of proteins in promastigote and amastigote-like Leishmania using an immunoproteomic approach. PLoS Neglected Tropical Diseases 6, e1430.
Costa, LE, Salles, BCS, Santos, TTO, Ramos, FF, Lima, MP, Lima, MIS, Portela, ASB, Chávez-Fumagalli, MA, Duarte, MC, Menezes-Souza, D, Machado-de-Ávila, RA, Silveira, JAG, Magalhães-Soares, DF, Goulart, LR and Coelho, EAF (2017) Antigenicity of phage clones and their synthetic peptides for the serodiagnosis of canine and human visceral leishmaniasis. Microbial Pathogenesis 110, 1422.
Costa, MM, Andrade, HM, Bartholomeu, DC, Freitas, LM, Pires, SF, Chapeaurouge, AD, Perales, J, Ferreira, AT, Giusta, MS, Melo, MN and Gazzinelli, RT (2011) Analysis of Leishmania chagasi by 2-D difference gel electrophoresis (2-D DIGE) and immunoproteomic: identification of novel candidate antigens for diagnostic tests and vaccine. Journal of Proteome Research 10, 21722184.
Duarte, MC, Lage, DP, Martins, VT, Costa, LE, Lage, LM, Carvalho, AM, Ludolf, F, Santos, TT, Roatt, BM, Menezes-Souza, D, Fernandes, AP, Tavares, CA and Coelho, EA (2016) A vaccine combining two Leishmania braziliensis proteins offers heterologous protection against Leishmania infantum infection. Molecular Immunology 76, 7079.
Duarte, MC, Lage, DP, Martins, VT, Costa, LE, Carvalho, AM, Ludolf, F, Santos, TT, Vale, DL, Roatt, BM, Menezes-Souza, D, Fernandes, AP, Tavares, CA and Coelho, EA (2017) A vaccine composed of a hypothetical protein and the eukaryotic initiation factor 5a from Leishmania braziliensis cross-protection against Leishmania amazonensis infection. Immunobiology 222, 251260.
Dupé, A, Dumas, C and Papadopoulou, B (2015) Differential subcellular localization of Leishmania alba-domain proteins throughout the parasite development. PLoS ONE 10, e0137243.
Eskandari, F, Talesh, GA, Parooie, M, Jaafari, MR, Khamesipour, A, Saberi, Z, Abbasi, A and Badiee, A (2014) Immunoliposomes containing soluble Leishmania antigens (SLA) as a novel antigen delivery system in murine model of leishmaniasis. Experimental Parasitology 146, 7886.
Faria, JA, Andrade, C, Goes, AM, Rodrigues, MA and Gomes, DA (2016) Effects of different ligands on epidermal growth factor receptor (EGFR) nuclear translocation. Biochemical and Biophysical Research Communication 478, 3945.
Fernandes, AP, Coelho, EAF, Machado-Coelho, GLL, Grimaldi, G Jr. and Gazzinelli, RT (2012) Making an anti-amastigote vaccine for visceral leishmaniasis: rational, update and perspectives. Current Opinion in Microbiology 15, 110.
Forestier, CL, Gao, Q and Boons, GJ (2015) Leishmania lipophosphoglycan: how to establish structure-activity relationships for this highly complex and multifunctional glycoconjugate? Frontiers in Cellular and Infection Microbiology 4, 193.
Gadisa, E, Custodio, E, Cañavate, C, Sordo, L, Abebe, Z, Nieto, J, Chicharro, C, Aseffa, A, Yamuah, L, Engers, H, Moreno, J and Cruz, I (2012) Usefulness of the rK39-immunochromatographic test, direct agglutination test, and leishmanin skin test for detecting asymptomatic Leishmania infection in children in a new visceral leishmaniasis focus in Amhara State, Ethiopia. The American Journal of Tropical Medicine and Hygiene 86, 792798.
Ghosh, P, Hasnain, MG, Ghosh, D, Hossain, F, Baker, J, Boelaert, M, Rijal, S and Mondal, D (2015) A comparative evaluation of the performance of commercially available rapid immunochromatographic tests for the diagnosis of visceral leishmaniasis in Bangladesh. Parasites & Vectors 8, 331.
Goto, Y, Bhatia, A, Raman, VS, Liang, H, Mohamath, R, Picone, AF, Vidal, SE, Vedvick, TS, Howard, RF and Reed, SG (2011) KSAC, the first defined polyprotein vaccine candidate for visceral leishmaniasis. Clinical and Vaccine Immunology 18, 11181124.
Green, LC, Wagner, DA, Glogowski, J, Skipper, PL, Wishnok, JS and Tannenbaum, SR (1982) Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids. Analytical Biochemistry 126, 131138.
Gupta, R, Kumar, V, Kushawaha, PK, Tripathi, CP, Joshi, S, Sahasrabuddhe, AA, Mitra, K, Sundar, S, Siddiqi, MI and Dube, A (2014) Characterization of glycolytic enzymes – rAldolase and rEnolase of Leishmania donovani, identified as Th1 stimulatory proteins, for their immunogenicity and immunoprophylactic efficacies against experimental visceral leishmaniasis. PLoS ONE 9, e86073.
Heung, LJ, Luberto, C and Del Poeta, M (2006) Role of sphingolipids in microbial pathogenesis. Infection and Immunity 74, 2839.
Iborra, S, Parody, N, Abanades, DR, Bonay, P, Prates, D, Novais, FO, Barral-Netto, M, Alonso, C and Soto, M (2008) Vaccination with the Leishmania major ribosomal proteins plus CpG oligodeoxynucleotides induces protection against experimental cutaneous leishmaniasis in mice. Microbes and Infection 10, 11331141.
Joshi, J and Kaur, S (2014) Studies on the protective efficacy of second-generation vaccine along with standard antileishmanial drug in Leishmania donovani infected BALB/c mice. Parasitology 141, 554562.
Kumari, S, Kumar, A, Samant, M, Sundar, S, Singh, N and Dube, A (2008) Proteomic approaches for discovery of new targets for vaccine and therapeutics against visceral leishmaniasis. Proteomics Clinical Applications 2, 372386.
Martins, VT, Lage, DP, Duarte, MC, Costa, LE, Garde, E, Rodrigues, MR, Chávez-Fumagalli, MA, Menezes-Souza, D, Roatt, BM, Tavares, CA, Soto, M and Coelho, EA (2016) A new Leishmania-specific hypothetical protein, LiHyT, used as a vaccine antigen against visceral leishmaniasis. Acta Tropica 154, 7381.
Martins, VT, Duarte, MC, Lage, DP, Costa, LE, Carvalho, AM, Mendes, TA, Roatt, BM, Menezes-Souza, D, Soto, M and Coelho, EA (2017) A recombinant chimeric protein composed of human and mice-specific CD4+ and CD8+ T-cell epitopes protects against visceral leishmaniasis. Parasite Immunology 39, e12359.
Matlashewski, G, Das, VN, Pandey, K, Singh, D, Das, S, Ghosh, AK, Pandey, RN and Das, P (2013) Diagnosis of visceral leishmaniasis in Bihar India: comparison of the rK39 rapid diagnostic test on whole blood versus serum. PLoS Neglected Tropical Diseases 7, e2233.
Miahipour, A, Haji-Fatahaliha, M, Keshavarz, H, Gharavi, MJ, Mohamadi, H, Babaloo, Z, Rafati, S, Younesi, V, Hosseini, M and Yousefi, M (2016) T helper 1 (Th1), Th2, and Th17 responses to Leishmania major lipophosphoglycan 3. Immunological Investigations 45, 692702.
Mizbani, A, Taheri, T, Zahedifard, F, Taslimi, Y, Azizi, H, Azadmanesh, K, Papadopoulou, B and Rafati, S (2009) Recombinant Leishmania tarentolae expressing the A2 virulence gene as a novel candidate vaccine against visceral leishmaniasis. Vaccine 28, 5362.
Mohamed-Ahmed, AH, Brocchini, S and Croft, SL (2012) Recent advances in development of amphotericin B formulations for the treatment of visceral leishmaniasis. Current Opinion in Infectious Diseases 25, 695702.
Mohammadiha, A, Mohebali, M, Haghighi, A, Mahdian, R, Abadi, AR, Zarei, Z, Yeganeh, F, Kazemi, B, Taghipour, N and Akhoundi, B (2013) Comparison of real-time PCR and conventional PCR with two DNA targets for detection of Leishmania (Leishmania) infantum infection in human and dog blood samples. Experimental Parasitology 133, 8994.
Peruhype-Magalhães, V, Martins-Filho, OA, Prata, A, Silva, LA, Rabello, A, Teixeira-Carvalho, A, Figueiredo, RM, Guimarães-Carvalho, SF, Ferrari, TC and Correa-Oliveira, R (2005) Immune response in human visceral leishmaniasis: analysis of the correlation between innate immunity cytokine profile and disease outcome. Scandinavian Journal of Immunology 62, 487495.
Petitdidier, E, Pagniez, J, Papierok, G, Vincendeau, P, Lemesre, JL and Bras-Gonçalves, R (2016) Recombinant forms of Leishmania amazonensis excreted/secreted promastigote surface antigen (PSA) induce protective immune responses in dogs. PLoS Neglected Tropical Diseases 10, e0004614.
Rafati, S, Kariminia, A, Seyde-Eslami, S, Narimani, M, Taheri, T and Lebbatard, M (2002) Recombinant cysteine proteinases-based vaccines against Leishmania major in BALB/c mice: the partial protection relies on interferon-gamma producing CD8(+) T lymphocyte activation. Vaccine 20, 24392447.
Reed, SG, Coler, RN, Mondal, D, Kamhawi, S and Valenzuela, JG (2016) Leishmania vaccine development: exploiting the host-vector-parasite interface. Expert Review of Vaccines 15, 8190.
Rodrigues, V, Cordeiro-da-Silva, A, Laforge, M, Silvestre, R and Estaquier, J (2016) Regulation of immunity during visceral Leishmania infection. Parasites & Vectors 9, 118.
Saldarriaga, OA, Travi, BL, Park, W, Perez, LE and Melby, PC (2006) Immunogenicity of a multicomponent DNA vaccine against visceral leishmaniasis in dogs. Vaccine 24, 19281940.
Saljoughian, N, Taheri, T, Zahedifard, F, Taslimi, Y, Doustdari, F, Bolhassani, A, Doroud, D, Azizi, H, Heidari, K, Vasei, M, Namvar, AN, Papadopoulou, B and Rafati, S (2013) Development of novel prime-boost strategies based on a tri-gene fusion recombinant L. tarentolae vaccine against experimental murine visceral leishmaniasis. PLoS Neglected Tropical Diseases 7, 115.
Singh, OP, Stober, CB, Singh, AK, Blackwell, JM and Sundar, S (2012) Cytokine responses to novel antigens in an Indian population living in an area endemic for visceral leishmaniasis. PLoS Neglected Tropical Diseases 6, e1874.
Srivastava, A, Singh, N, Mishra, M, Kumar, V, Gour, JK, Bajpai, S, Singh, S, Pandey, HP and Singh, RK (2012) Identification of TLR inducing Th1-responsive Leishmania donovani amastigote-specific antigens. Molecular and Cellular Biochemistry 359, 359368.
Stockdale, L and Newton, R (2013) A review of preventative methods against human leishmaniasis infection. PLoS Neglected Tropical Diseases 7, e2278.
Sundar, S and Chakravarty, J (2013) Leishmaniasis: an update of current pharmacotherapy. Expert Opinion on Pharmacotherapy 14, 5363.
Sundar, S and Singh, B (2014) Identifying vaccine targets for anti-leishmanial vaccine development. Expert Review of Vaccines 13, 489505.
Thakur, A, Kaur, H and Kaur, S (2015) Studies on the protective efficacy of freeze thawed promastigote antigen of Leishmania donovani along with various adjuvants against visceral leishmaniasis infection in mice. Immunobiology 220, 10311038.
Vieira, PM, Francisco, AF, Machado, EM, Nogueira, NC, Fonseca, KS, Reis, AB, Teixeira-Carvalho, A, Martins-Filho, OA, Tafuri, WL and Carneiro, CM (2012) Different infective forms trigger distinct immune response in experimental Chagas disease. PLoS ONE 7, e32912.
Yao, C, Li, Y, Donelson, JE and Wilson, ME (2010) Proteomic examination of Leishmania chagasi plasma membrane proteins: contrast between avirulent and virulent (metacyclic) parasite forms. Proteomics Clinical Applications 4, 416.
Zadeh-Vakili, A, Taheri, T, Taslimi, Y, Doustdari, F, Salmanian, AH and Rafati, S (2004) Immunization with the hybrid protein vaccine, consisting of Leishmania major cysteine proteinases Type I (CPB) and Type II (CPA), partially protects against leishmaniasis. Vaccine 22, 19301940.
Zhang, WW, Charest, H, Ghedin, E and Matlashewski, G (1996) Identification and overexpression of the A2 amastigote-specific protein in Leishmania donovani. Molecular and Biochemical Parasitology 78, 7990.
Werbovetz, KA, Brendle, JJ and Sackett, DL (1999) Purification, characterization, and drug susceptibility of tubulin from Leishmania. Molecular and Biochemical Parasitology 98, 5365.

Keywords

Type Description Title
WORD
Supplementary materials

Dias et al supplementary material 1
Supplementary Figure

 Word (358 KB)
358 KB

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed