Sequencing and Gene Expression Analysis of Leishmania tropica LACK Gene.

  • Nour Hammoudeh Dept. of Biotechnology, Faculty of Agriculture, Damascus University, Damascus, Syria.
  • Mahmoud Kweider Dept. of Animal Biology, Faculty of Sciences, Damascus University, Damascus, Syria.
  • Abdul-Qader Abbady Dept. of Molecular Biology and Biotechnology, Atomic Energy Commission of Syria(AECS), Damascus, Syria.
  • Chadi Soukkarieh Dept. of Animal Biology, Faculty of Sciences, Damascus University, Damascus, Syria.
Keywords: Amastigotes, Cutaneous leishmaniasis, LACK gene, Leishmania tropica, Promastigotes


Background: Leishmania Homologue of receptors for Activated C Kinase(LACK) antigen is a 36-kDa protein, which provokes a very early immune response against Leishmania infection. There are several reports on the expression of LACK through different life-cycle stages of genus Leishmania, but only a few of them have focused on L.tropica.Methods: The present study provides details of the cloning, DNA sequencing and gene expression of LACK in this parasite species. First,several local isolates of Leishmania parasites were typed in our laboratory using PCR technique to verify of Leishmania parasite species. After that,LACK gene was amplified and cloned into a vector for sequencing. Finally,the expression of this molecule in logarithmic and stationary growth phase promastigotes, as well as in amastigotes, was evaluated by Reverse Transcription-PCR (RT-PCR) technique.Results: The typing result confirmed that all our local isolates belong to L.tropica. LACK gene sequence was determined and high similarity was observed with the sequences of other Leishmania species. Furthermore,the expression of LACK gene in both promastigotes and amastigotesforms was confirmed.Conclusion: Overall, the data set the stage for future studies of the properties and immune role of LACK gene products.


WHO. Report of the consultative meeting on cutaneous leishmaniasis word health organization. 2007:1-36.

Dunning N. Leishmania vaccines: From leishmanization to the era of DNA technology. Bioscience Horizons. 2009;2:73-82.

Pearson RD, Sousa AQ. Clinical spectrum of leishmaniasis. Clin Infect Dis. 1996;22:1-13.

Chappuis F, Sundar S, Hailu A, Ghalib H, Rijal S, Peeling RW, Alvar J, Boelaert M. Visceral leishmaniasis: What are the needs for diagnosis, treatment and control? Nat Rev Microbiol. 2007;5:873-882.

Odiwuor SO, Saad AA, De Doncker S, Maes I, Laurent T, El Safi S, Mbuchi M, Buscher P, Dujardin JC, Van der Auwera G. Universal PCR assays for the differential detection of all old world Leishmania species. Eur J Clin Microbiol Infect Dis. 2011;30:209-218.

Olivier M, Gregory DJ, Forget G. Subversion mechanisms by which Leishmania parasites can escape the host immune response: A signaling point of view. Clin Microbiol Rev. 2005;18:293-305.

Coelho EA, Tavares CA, Carvalho FA, Chaves KF, Teixeira KN, Rodrigues RC, Charest H, Matlashewski G, Gazzinelli RT, Fernandes AP. Immune responses induced by the Leishmania(Leishmania) donovani A2 antigen, but not by the LACK antigen, are protective against experimental Leishmania (Leishmania) amazonensis infection. Infect Immun. 2003;71:3988-3994.

Kelly BL, Stetson DB, Locksley RM. Leishmania major LACK antigen is required for efficient vertebrate parasitization. J Exp Med. 2003;198:1689-1698.

Gomez-Arreaza A, Acosta H, Barros-Alvarez X, Concepcion JL, Albericio F, Avilan L. Leishmania mexicana: LACK (leishmania homolog of receptors for activated c-kinase) is a plasminogen binding protein. Exp Parasitol. 2011;127:752-761.

Melby PC, Yang J, Zhao W, Perez LE, Cheng J. Leishmania donovani p36(LACK) DNA vaccine is highly immunogenic but not protective against experimental visceral leishmaniasis. Infect Immun. 2001;69:4719-4725.

Okuno T, Takeuchi M, Matsumoto Y, Otsuka H. Pretreatment of Leishmania homologue of receptors for activated c kinase (LACK) promotes disease progression caused by Leishmania amazonensis. Exp Anim. 2002;51:335-341.

Kubar J, Fragaki K. Leishmania proteins derived from recombinant DNA: Current status and next steps. Trends Parasitol. 2006;22:111-116.

Mugasa CM, Laurent T, Schoone GJ, Basiye FL, Saad AA, El Safi S, Kager PA, Schallig HD. Simplified molecular detection of Leishmania parasites in various clinical samples from patients with leishmaniasis. Parasit Vectors. 2010;3:13.

Chulay JD, Bryceson AD. Quantitation of amastigotes of Leishmania donovani in smears of splenic aspirates from patients with visceral leishmaniasis. Am J Trop Med Hyg. 1983; 32:475-479.

Sereno D, Lemesre JL. In vitro life cycle of pentamidine-resistant amastigotes: Stability of the chemoresistant phenotypes is dependent on the level of resistance induced. Antimicrob Agents Chemother. 1997;41:1898-1903.

El Fakhry Y, Ouellette M, Papadopoulou B. A proteomic approach to identify developmentally regulated proteins in Leishmania infantum. Proteomics. 2002;2:1007-1017.

Barak E, Amin-Spector S, Gerliak E, Goyard S, Holland N, Zilberstein D. Differentiation of Leishmania donovani in host-free system: Analysis of signal perception and response. Mol Biochem Parasitol. 2005;141:99-108.

Saar Y, Ransford A, Waldman E, Mazareb S, Amin-Spector S, Plumblee J, Turco SJ, Zilberstein D. Characterization of developmentally-regulated activities in axenic amastigotes of Leishmania donovani. Mol Biochem Parasitol. 1998;95:9-20.

Ray DS. Conserved sequence blocks in kinetoplast minicircles from diverse species of trypanosomes. Mol Cell Biol. 1989;9:1365-1367.

Yurchenko VY, Merzlyak EM, Kolesnikov AA, Martinkina LP, Vengerov YY. Structure of Leishmania minicircle kinetoplast DNA classes. J Clin Microbiol. 1999;37:1656-1657.

Gomes AH, Ferreira IM, Lima ML, Cunha EA, Garcia AS, Araujo MF, Pereira-Chioccola VL. Pcr identification of Leishmania in diagnosis and control of canine leishmaniasis. Vet Parasitol. 2007;144:234-241.

Bensoussan E, Nasereddin A, Jonas F, Schnur LF, Jaffe CL. Comparison of pcr assays for diagnosis of cutaneous leishmaniasis. J Clin Microbiol. 2006;44:1435-1439.

Courret N, Prina E, Mougneau E, Saraiva EM, Sacks DL, Glaichenhaus N, Antoine JC. Presentation of the Leishmania antigen LACK by infected macrophages is dependent upon the virulence of the phagocytosed parasites. Eur J Immunol. 1999;29:762-773.

Hajjaran H, Azarian B, Mohebali M, Hadighi R, Assareh A, Vaziri B. Comparative proteomics study on meglumine antimoniate sensitive and resistant Leishmania tropica isolated from iranian anthroponotic cutaneous leishmaniasis patients. Eastern Mediterranean health journal = La revue de sante de la Mediterranee orientale = al-Majallah al-sihhiyah li-sharq al-mutawassit. 2012;18:165-171.

Maasho K, Satti I, Nylen S, Guzman G, Koning F, Akuffo H. A Leishmania homologue of receptors for activated c-kinase (LACK) induces both interferon-gamma and interleukin-10 in natural killer cells of healthy blood donors. J Infect Dis. 2000;182:570-578.

Mougneau E, Altare F, Wakil AE, Zheng S, Coppola T, Wang ZE, Waldmann R, Locksley RM, Glaichenhaus N. Expression cloning of a protective Leishmania antigen. Science. 1995;268:563-566.

Gurunathan S, Sacks DL, Brown DR, Reiner SL, Charest H, Glaichenhaus N, Seder RA. Vaccination with DNA encoding the immunodominant LACK parasite antigen confers protective immunity to mice infected with Leishmania major. J Exp Med. 1997;186:1137-1147.

Kramer S. Developmental regulation of gene expression in the absence of transcriptional control: The case of kinetoplastids. Mol Biochem Parasitol. 2012;181:61-72.

How to Cite
Hammoudeh N, Kweider M, Abbady A-Q, Soukkarieh C. Sequencing and Gene Expression Analysis of Leishmania tropica LACK Gene. IJPA. 9(4):574-83.