Recombinant LPG3 Stimulates IFN-Γ and TNF-Α Secretion by Human NK Cells
-
Sanaz RASOLZADEH
,
Mostafa HAJI FATAHALIHA
,
Maryam HOSSEINI
,
Reza JAFARI
,
Abolfazl MIAHIPOUR
,
Ali Akbar MOVASSAGHPOUR
,
Zohreh BABALO
,
Sima RAFATI
,
Mehdi YOUSEFI
Abstract
Background: Natural killer (NK) cells play an important role in early stages of innate immune responses against viral and tumoral attacks. Activation of NK cells by leishmaniasis results in secretion of cytokines such as interferon (IFN)-γ and tumor necrosis factor (TNF)-α, which enhances the phagocytosis and clearance of parasite. Lipophosphoglycan 3 (LPG3), the Leishmania homologous with GRP94 (glucose regulated protein 94), a member of HSP90 family, contributes to LPG assembly as the most abundant macromolecule on the surface of Leishmania promastigotes.
Methods: We purified NK cells from healthy individuals (n=10) using magnetic-activated cell sorting (MACS) technology. Purified NK cells were co-incubated with different concentrations of recombinant LPG3 (rLPG3), and its N-terminal (NT) and C-terminal (CT) fragments. Finally, the production of IFN-γ and TNF-α by NK cells were measured by ELISA.
Results: Recombinant LPG3 but not its fragments (CT and NT), can significantly enhance the production of TNF-α by NK cells (P<0.05). Moreover, rLPG3, CT, and NT fragments were markedly stimulated the secretion of IFN-γ by NK cells (P<0.001).
Conclusion: The Leishmania LPG3 antigen can effectively activate NK cells, in vitro. Leishmania LPG3 participates in the innate immunity against leishmaniasis and thereby improves the effective parasite destruction. However, its efficiency should be tested in vivo.Alvar J, Velez ID, Bern C, Herrero M, Desjeux P, Cano J, Jannin J, den Boer M, Team WLC. Leishmaniasis worldwide and global estimates of its incidence. PloS One. 2012;7:e35671.
Descoteaux A, Avila HA, Zhang K, Turco SJ, Beverley SM. Leishmania lpg3 encodes a grp94 homolog required for phosphoglycan synthesis implicated in parasite virulence but not viability. The EMBO J. 2002;21:4458-4469.
Franco LH, Beverley SM, Zamboni DS. Innate immune activation and subversion of mammalian functions by Leishmania lipophosphoglycan. J Parasitol Res. 2012;2012. Article ID 165126, 11 pages.
Joshi S, Rawat K, Yadav NK, Kumar V, Siddiqi MI, Dube A. Visceral leishmaniasis: Advanceme-nts in vaccine development via classical and molecular approaches. Frontiers Immunol. 2014;5.
Seyed N, Taheri T, Vauchy C, Dosset M, Godet Y, Eslamifar A, Sharifi I, Adotevi O, Borg C, Rohrlich PS. Immunogenicity evaluation of a rationally designed polytope construct encoding hla-a* 0201 restricted epitopes derived from leishmania major related proteins in hla-a2/dr1 transgenic mice: Steps toward polytope vaccine. PloS One. 2014;9:e108848.
Reithinger R, Dujardin J-C, Louzir H, Pirmez C, Alexander B, Brooker S. Cutaneous leishmaniasis. The Lancet Infect Dis. 2007;7:581-596.
Beverley SM, Turco SJ. Lipophosphoglycan (lpg) and the identification of virulence genes in the protozoan parasite Leishmania. Trends Microbiol. 1998;6:35-40.
Ilg T. Lipophosphoglycan is not required for infection of macrophages or mice by Leishmania mexicana. EMBO J. 2000;19:1953-1962.
McConville M, Schnur L, Jaffe C, Schneider P. Structure of Leishmania lipophosphoglycan: Inter-and intra-specific polymorphism in old world species. Biochem J. 1995;310:807-818.
Croft SL, Barrett MP, Urbina JA. Chemoth-erapy of trypanosomiases and leishmaniasis. Trends Parasitol. 2005;21:508-512.
Lanier LL. Turning on natural killer cells. J Exp Med. 2000;191:1259-1262.
Liese J, Schleicher U, Bogdan C. The innate immune response against Leishmania parasites. Immunobiology. 2008;213:377-387.
Scott P, Trinchieri G. The role of natural killer cells in host—parasite interactions. Current Opinion Immunol. 1995;7:34-40.
Akuffo H, Maasho K, Howe R. Natural and acquired resistance to Leishmania: Cellular activation by Leishmania aethiopica of mononuclear cells from unexposed individuals is through the stimulation of natural killer (NK) cells. Clin Exp Immunol. 1993;94:516-521.
Abdian N, Gholami E, Zahedifard F, Safaee N, Rafati S. Evaluation of DNA/DNA and prime-boost vaccination using lpg3 against Leishmania major infection in susceptible balb/c mice and its antigenic properties in human leishmaniasis. Exp Parasitol. 2011;127:627-636.
Laskay T, Diefenbach A, Röllinghoff M, Solbach W. Early parasite containment is decisive for resistance to Leishmania major infection. Eur J Immunol. 1995;25:2220-2227.
Maasho K, Sanchez F, Schurr E, Hailu A, Akuffo H. Indications of the protective role of natural killer cells in human cutaneous leishmaniasis in an area of endemicity. Infect Immun. 1998;66:2698-2704.
Vester B, Müller K, Solbach W, Laskay T. Early gene expression of nk cell-activating chemokines in mice resistant to Leishmania major. Infect Immun. 1999;67:3155-3159.
Salaiza‐Suazo N, Volkow P, Tamayo R, Moll H et al.. Treatment of two patients with diffuse cutaneous leishmaniasis caused by Leishmania mexicana modifies the immunoh-istological profile but not the disease outcome. Trop Med Int Health. 1999;4:801-811.
Pinheiro RO, Rossi-Bergmann B. Interferon-gamma is required for the late but not early control of Leishmania amazonensis infection in c57bl/6 mice. Memórias do Instituto Oswaldo Cruz. 2007;102:79-82.
Swihart K, Fruth U, Messmer N, Hug K, Behin R, Huang S, Del Giudice G, Aguet M, Louis JA. Mice from a genetically resistant background lacking the interferon γ receptor are susceptible to infection with Leishmania major but mount a polarized t helper cell 1-type cd4+ t cell response. J Exp Med. 1995;181:961-971.
Moretta L, Biassoni R, Bottino C, Mingari MC, Moretta A. Human NK-cell receptors. Immunol Today. 2000;21:420-422.
Stepp SE, Mathew PA, Bennett M, de Saint Basile G, Kumar V. Perforin: More than just an effector molecule. Immunol Today. 2000;21:254-256.
Theodos C, Povinelli L, Molina R, Sherry B, Titus R. Role of tumor necrosis factor in macrophage leishmanicidal activity in vitro and resistance to cutaneous leishmaniasis in vivo. Infect Immun. 1991;59:2839-2842.
Liew F, Li Y, Millott S. Tumor necrosis factor-alpha synergizes with IFN-gamma in mediating killing of Leishmania major through the induction of nitric oxide. J Immunol. 1990;145:4306-4310.
Liew F, Parkinson C, Millott S, Severn A, Carrier M. Tumour necrosis factor (TNF alpha) in leishmaniasis. I. TNF alpha mediates host protection against cutaneous leishmaniasis. Immunology. 1990;69:570.
De Kossodo S, Grau GE, Louis JA, Müller I. Tumor necrosis factor alpha (TNF-alpha) and tnf-beta and their receptors in experimental cutaneous leishmaniasis. Infect Immun. 1994;62:1414-1420.
Garcia I, Miyazaki Y, Araki K, Araki M, Lucas R, Grau GE, Milon G, Belkaid Y, Montixi C, Lesslauer W. Transgenic mice expressing high levels of soluble TNF‐r1 fusion protein are protected from lethal septic shock and cerebral malaria, and are highly sensitive to Listeria monocytogenes and Leishmania major infections. Eur J Immunol. 1995;25:2401-2407.
Bogdan C, Moll H, Solbach W, Röllinghoff M. Tumor necrosis factor‐α in combination with interferon‐γ, but not with interleukin 4 activates murine macrophages for elimination of Leishmania major amastigotes. Eur J Immunol. 1990;20:1131-1135.
Bogdan C, Röllinghoff M, Diefenbach A. Reactive oxygen and reactive nitrogen intermediates in innate and specific immunity. Current Opinion Immunol. 2000;12:64-76.
Bogdan C, Röllinghoff M, Diefenbach A. The role of nitric oxide in innate immunity. Immunol Rev. 2000;173:17-26.
Reiner SL, Locksley RM. The regulation of immunity to Leishmania major. Ann Rev Immunol. 1995;13:151-177.
Pirdel L, Zavaran Hosseini A, Rasouli M. Immune response in susceptible balb/c mice immunized with DNA encoding lipophospho-glycan 3 of Leishmania infantum. Parasite Immunol. 2014;36:700-707.
| Files | ||
| Issue | Vol 10 No 4 (2015) | |
| Section | Original Article(s) | |
| Keywords | ||
| Leishmania LPG3 Natural killer cell TLR | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
