Skip to main content

Advertisement

SpringerLink
Log in

Immunization against leishmaniasis by PLGA nanospheres encapsulated with autoclaved Leishmania major (ALM) and CpG-ODN

  • Original Paper
  • Published:
Parasitology Research Aims and scope Submit manuscript

Abstract

Various adjuvants and delivery systems have been evaluated for increasing the protective immune responses against leishmaniasis and mostly have been shown not to be effective enough. In this study, poly(d,l-lactide-co-glycolide) (PLGA) nanospheres as an antigen delivery system and CpG-ODN as an immunoadjuvant have been used for the first time to enhance the immune response against autoclaved Leishmania major (ALM). PLGA nanospheres were prepared by a double-emulsion (W/O/W) technique. Particulate characteristics were studied by scanning electron microscopy and particle size analysis. Mean diameter of ALM + CpG-ODN-loaded nanospheres was 300 ± 128 nm. BALB/c mice were immunized three times in 3-week intervals using ALM plus CpG-ODN-loaded nanospheres [(ALM + CpG-ODN)PLGA], ALM encapsulated PLGA nanospheres [(ALM)PLGA], (ALM)PLGA + CpG, ALM + CpG, ALM alone, or phosphate buffer solution (PBS). The intensity of infection induced by L. major challenge was assessed by measuring size of footpad swelling. The strongest protection, showed by significantly (P < 0.05) smaller footpad, was observed in mice immunized with (ALM + CpG-ODN)PLGA. The (ALM)PLGA, (ALM)PLGA + CpG, and ALM + CpG were also showed a significantly (P < 0.05) smaller footpad swelling compared to the groups received either PBS or ALM alone. The mice immunized with (ALM + CpG-ODN)PLGA, (ALM)PLGA + CpG, and ALM + CpG showed the highest IgG2a/IgG1 ratio, interferon-γ production, and lowest interleukin-4 production compared to the other groups. It is concluded that when both PLGA nanospheres and CpG-ODN adjuvants were used simultaneously, it induce stronger immune response and enhance protection rate against Leishmania infection.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Ahmad-Nejad P, Hacker H, Rutz M, Bauer S, Vabulas RM, Wagner H (2002) Bacterial CPG-DNA and lipopolysaccharides activate toll-like receptors at distinct cellular compartments. Eur J Immunol 32(7):1958–1968

    Article  PubMed  CAS  Google Scholar 

  • Alimohammadian MH, Khamesipour A, Darabi H, Firooz A, Malekzadeh S, Bahonar A, Dowlati Y, Modabber F (2002) The role of BCG in human immune responses induced by multiple injections of autoclaved Leishmania major as a candidate vaccine against leishmaniasis. Vaccine 21(3–4):174–180

    Article  PubMed  CAS  Google Scholar 

  • Armijos RX, Weigel MM, Calvopina M, Hidalgo A, Cevallos W, Correa J (2004) Safety, immunogenecity, and efficacy of an autoclaved Leishmania amazonensis vaccine plus BCG adjuvant against new world cutaneous leishmaniasis. Vaccine 22(9–10):1320–1326

    Article  PubMed  CAS  Google Scholar 

  • Badiee A, Jaafari MR, Khamesipour A (2007) Leishmania major: immune response in BALB/c mice immunized with stress-inducible protein 1 encapsulated in liposomes. Exp Parasitol 115(2):127–134

    Article  PubMed  CAS  Google Scholar 

  • Bahar K, Dowlati Y, Shidani B, Alimohammadian MH, Khamesipour A, Ehsasi S, Hashemi-Fesharki R, Ale-Agha S, Modabber F (1996) Comparative safety and immunogenicity trial of two killed Leishmania major vaccines with or without BCG in human volunteers. Clin Dermatol 14(5):489–495

    Article  PubMed  CAS  Google Scholar 

  • Barman SP, Lunsford L, Chambers P, Hedley ML (2000) Two methods for quantifying DNA extracted from poly(lactide-co-glycolide) microspheres. J Control Release 69(3):337–344

    Article  PubMed  CAS  Google Scholar 

  • Bhowmick S, Ravindran R, Ali N (2007) Leishmanial antigens in liposomes promote protective immunity and provide immunotherapy against visceral leishmaniasis via polarized th1 response. Vaccine 25(35):6544–6556

    Article  PubMed  CAS  Google Scholar 

  • Cabrera M, Blackwell JM, Castes M, Trujillo D, Convit J, Shaw MA (2000) Immunotherapy with live BCG plus heat killed Leishmania induces a T helper 1-like response in American cutaneous leishmaniasis patients. Parasite Immunol 22(2):73–79

    Article  PubMed  CAS  Google Scholar 

  • Coelho EA, Tavares CA, Lima Kde M, Silva CL, Rodrigues JM Jr, Fernandes AP (2006) Mycobacterium HSP65 DNA entrapped into TDM-loaded PLGA microspheres induces protection in mice against Leishmania (leishmania) major infection. Parasitol Res 98(6):568–575

    Article  PubMed  Google Scholar 

  • De Luca PM, Mayrink W, Alves CR, Coutinho SG, Oliveira MP, Bertho AL, Toledo VP, Costa CA, Genaro O, Mendonca SCF (1999) Evaluation of the stability and immunogenicity of autoclaved and nonautoclaved preparations of a vaccine against American tegumentary leishmaniasis. Vaccine 17(9–10):1179–1185

    PubMed  Google Scholar 

  • Diwan M, Tafaghodi M, Samuel J (2002) Enhancement of immune responses by co-delivery of a CPG oligodeoxynucleotide and tetanus toxoid in biodegradable nanospheres. J Control Release 85(1–3):247–262

    Article  PubMed  CAS  Google Scholar 

  • Flynn B, Wang V, Sacks DL, Seder RA, Verthelyi D (2005) Prevention and treatment of cutaneous leishmaniasis in primates by using synthetic type D/A oligodeoxynucleotides expressing CPG motifs. Infect Immun 73(8):4948–4954

    Article  PubMed  CAS  Google Scholar 

  • Follador I, Araujo C, Orge G, Cheng LH, de Carvalho LP, Bacellar O, Almeida RP, Carvalho EM (2002) Immune responses to an inactive vaccine against American cutaneous leishmaniasis together with granulocyte-macrophage colony-stimulating factor. Vaccine 20(9–10):1365–1368

    Article  PubMed  CAS  Google Scholar 

  • Gupta RK, Siber GR (1995) Adjuvants for human vaccines—current status, problems and future prospects. Vaccine 13(14):1263–1276

    Article  PubMed  CAS  Google Scholar 

  • Hemmi H, Takeuchi O, Kawai T, Kaisho T, Sato S, Sanjo H, Matsumoto M, Hoshino K, Wagner H, Takeda K et al (2000) A toll-like receptor recognizes bacterial DNA. Nature 408(6813):740–745

    Article  PubMed  CAS  Google Scholar 

  • Iborra S, Carrion J, Anderson C, Alonso C, Sacks D, Soto M (2005) Vaccination with the Leishmania infantum acidic ribosomal p0 protein plus CPG oligodeoxynucleotides induces protection against cutaneous leishmaniasis in c57bl/6 mice but does not prevent progressive disease in BALB/c mice. Infect Immun 73(9):5842–5852

    Article  PubMed  CAS  Google Scholar 

  • Iborra S, Parody N, Abánades DR, Bonay P, Prates D, Novais FO, Barral-Netto M, Alonso C, Soto M (2008) Vaccination with the Leishmania major ribosomal proteins plus CPG oligodeoxynucleotides induces protection against experimental cutaneous leishmaniasis in mice. Microbes Infect 10:1133–1141

    Article  PubMed  CAS  Google Scholar 

  • Jaafari MR, Ghafarian A, Farrokh-Gisour A, Samiei A, Kheiri MT, Mahboudi F, Barkhordari F, Khamesipour A, McMaster WR (2006) Immune response and protection assay of recombinant major surface glycoprotein of Leishmania (rgp63) reconstituted with liposomes in BALB/c mice. Vaccine 24(29–30):5708–5717

    Article  PubMed  CAS  Google Scholar 

  • Jaafari MR, Badiee A, Khamesipour A, Samiei A, Soroush D, Kheiri MT, Barkhordari F, McMaster WR, Mahboudi F (2007) The role of CpG ODN in enhancement of immune response and protection in BALB/c mice immunized with recombinant major surface glycoprotein of Leishmania (rgp63) encapsulated in cationic liposome. Vaccine 25(32):6107–6117

    Article  PubMed  CAS  Google Scholar 

  • Javadian E, Nadim A, Tahvildare-Bidruni G, Assefi V (1976) Epidemiology of cutaneous leishmaniasis in Iran: B. Khorassan part V: report on a focus of zoonotic cutaneous leishmaniasis in Esferayen. Bull Soc Pathol Exot Fil 69(2):140–143

    CAS  Google Scholar 

  • Johansen P, Men Y, Merkle HP, Gander B (2000) Revisiting PLA/PLGA microspheres: an analysis of their potential in parenteral vaccination. Eur J Pharm Biopharm 50(1):129–146

    Article  PubMed  CAS  Google Scholar 

  • Kamil AA, Khalil EA, Musa AM, Modabber F, Mukhtar MM, Ibrahim ME, Zijlstra EE, Sacks D, Smith PG, Zicker F, El-Hassan AM (2003) Alum-precipitated autoclaved Leishmania major plus Bacille Calmette-Guerrin, a candidate vaccine for visceral leishmaniasis: safety, skin-delayed type hypersensitivity response and dose finding in healthy volunteers. Trans R Soc Trop Med Hyg 97(3):365–368

    Article  PubMed  CAS  Google Scholar 

  • Kedzierski L, Zhu Y, Handman E (2006) Leishmania vaccines: progress and problems. Parasitology 133(Suppl):S87–S112

    Article  PubMed  CAS  Google Scholar 

  • Keshavarz Valian H, Khoshabe Abdollah Kenedy L, Nateghi Rostami M, Miramin Mohammadi A, Khamesipour A (2008) Role of Mycobacterium vaccae in the protection induced by first generation Leishmania vaccine against murine model of leishmaniasis. Parasitol Res 103:21–28

    Article  PubMed  Google Scholar 

  • Khalil EA, El Hassan AM, Zijlstra EE, Mukhtar MM, Ghalib HW, Musa B, Ibrahim ME, Kamil AA, Elsheikh M, Babiker A et al (2000) Autoclaved Leishmania major vaccine for prevention of visceral leishmaniasis: a randomised, double-blind, BCG-controlled trial in Sudan. Lancet 356(9241):1565–1569

    Article  PubMed  CAS  Google Scholar 

  • Khamesipour A, Dowlati Y, Asilian A, Hashemi-Fesharki R, Javadi A, Noazin S, Modabber F (2005) Leishmanization: use of an old method for evaluation of candidate vaccines against leishmaniasis. Vaccine 23(28):3642–3648

    Article  PubMed  CAS  Google Scholar 

  • Khamesipour A, Rafati S, Davoudi N, Maboudi F, Modabber F (2006) Leishmaniasis vaccine candidates for development: a global overview. Indian J Med Res 123(3):423–438

    PubMed  Google Scholar 

  • Krieg AM (1999) Mechanisms and applications of immune stimulatory CPG oligodeoxynucleotides. Biochim Biophys Acta BBA Gene Struct Expr 1489(1):107–116

    CAS  Google Scholar 

  • Krieg AM (2002) CPG motifs in bacterial DNA and their immune effects. Annu Rev Immunol 20:709–760

    Article  PubMed  CAS  Google Scholar 

  • Mauel J (2002) Vaccination against Leishmania infections. Curr Drug Targets Immune Endocr Metabol Disord 2(3):201–226

    Article  PubMed  CAS  Google Scholar 

  • McCluskie MJ, Davis HL (1998) CPG DNA is a potent enhancer of systemic and mucosal immune responses against hepatitis B surface antigen with intranasal administration to mice. J Immunol (1950) 161(9):4463–4466

    CAS  Google Scholar 

  • Mendez S, Tabbara K, Belkaid Y, Bertholet S, Verthelyi D, Klinman D, Seder RA, Sacks DL (2003) Coinjection with CPG-containing immunostimulatory oligodeoxynucleotides reduces the pathogenicity of a live vaccine against cutaneous leishmaniasis but maintains its potency and durability. Infect Immun 71(9):5121–5129

    Article  PubMed  CAS  Google Scholar 

  • Misra A, Dube A, Srivastava B, Sharma P, Srivastava JK, Katiyar JC, Naik S (2001) Successful vaccination against Leishmania donovani infection in Indian langur using alum-precipitated autoclaved Leishmania major with BCG. Vaccine 19(25–26):3485–3492

    Article  PubMed  CAS  Google Scholar 

  • Modabber F (1995) Vaccines against leishmaniasis. Ann Trop Med Parasitol 89(Supplement 1):83–88

    PubMed  CAS  Google Scholar 

  • Mohebali M, Khamesipour A, Mobedi I, Zarei Z, Hashemi-Fesharki R (2004) Double-blind randomized efficacy field trial of alum precipitated autoclaved Leishmania major vaccine mixed with BCG against canine visceral leishmaniasis in Meshkin-Shahr District, I.R. Iran. Vaccine 22(29–30):4097–4100

    Article  PubMed  CAS  Google Scholar 

  • Newman KD, Sosnowski DL, Kwon GS, Samuel J (1998) Delivery of muc1 mucin peptide by poly(d, l-lactic-co-glycolic acid) microspheres induces type 1 T helper immune responses. J Pharm Sci 87(11):1421–1427

    Article  PubMed  CAS  Google Scholar 

  • Newman KD, Elamanchili P, Kwon GS, Samuel J (2002) Uptake of poly(d, l-lactic-co-glycolic acid) microspheres by antigen-presenting cells in vivo. J Biomed Mater Res 60(3):480–486

    Article  PubMed  CAS  Google Scholar 

  • Noazin S, Modabber F, Khamesipour A, Smith PG, Moulton LH, Nasseri K, Sharifi I, Khalil EA, Bernal ID, Antunes CM, Kieny MP, Tanner M (2008) First generation leishmaniasis vaccines: a review of field efficacy trials. Vaccine 26(52):6759–6767

    Article  PubMed  CAS  Google Scholar 

  • Rebelatto MC, Guimond P, Bowersock TL, HogenEsch H (2001) Induction of systemic and mucosal immune response in cattle by intranasal administration of pig serum albumin in alginate microparticles. Vet Immunol Immunopathol 83(1–2):93–105

    Article  PubMed  CAS  Google Scholar 

  • Reiner SL, Locksley RM (1995) The regulation of immunity to Leishmania major. Annu Rev Immunol 13:151–177

    Article  PubMed  CAS  Google Scholar 

  • Rhee EG, Mendez S, Shah JA, Wu CY, Kirman JR, Turon TN, Davey DF, Davis H, Klinman DM, Coler RN, Sacks DL, Seder RA (2002) Vaccination with heat-killed Leishmania antigen or recombinant leishmanial protein and CPG oligodeoxynucleotides induces long-term memory cd4+ and cd8+ T cell responses and protection against Leishmania major infection. J Exp Med 195(12):1565–1573

    Article  PubMed  CAS  Google Scholar 

  • Sacks D, Noben-Trauth N (2002) The immunology of susceptibility and resistance to Leishmania major in mice. Nat Rev Immunol 2(11):845–858

    Article  PubMed  CAS  Google Scholar 

  • Satti IN, Osman HY, Daifalla NS, Younis SA, Khalil EA, Zijlstra EE, El Hassan AM, Ghalib HW (2001) Immunogenicity and safety of autoclaved Leishmania major plus BCG vaccine in healthy Sudanese volunteers. Vaccine 19(15–16):2100–2106

    Article  PubMed  CAS  Google Scholar 

  • Scott P, Pearce E, Natovitz P, Sher A (1987) Vaccination against cutaneous leishmaniasis in a murine model. I. Induction of protective immunity with a soluble extract of promastigotes. J Immunol 139(1):221–227

    PubMed  CAS  Google Scholar 

  • Shimizu Y, Yamakami K, Gomi T, Nakata M, Asanuma H, Tadakuma T, Kojima N (2003) Protection against Leishmania major infection by oligomannose-coated liposomes. Bioorg Med Chem 11(7):1191–1195

    Article  PubMed  CAS  Google Scholar 

  • Shimizu Y, Takagi H, Nakayama T, Yamakami K, Tadakuma T, Yokoyama N, Kojima N (2007) Intraperitoneal immunization with oligomannose-coated liposome-entrapped soluble leishmanial antigen induces antigen-specific T-helper type immune response in BALB/c mice through uptake by peritoneal macrophages. Parasite Immunol 29(5):229–239

    Article  PubMed  CAS  Google Scholar 

  • Sinha VR, Trehan A (2003) Biodegradable microspheres for protein delivery. J Control Release 90(3):261–280

    Article  PubMed  CAS  Google Scholar 

  • Tafaghodi M, Sajadi Tabassi SA, Jaafari M-R, Zakavi SR, Momen Nejad M (2004) Evaluation of the clearance characteristics of various microspheres in the human nose by gamma-scintigraphy. Int J Pharm 280:125–135

    Article  PubMed  CAS  Google Scholar 

  • Tewary P, Pandya J, Mehta J, Sukumaran B, Madhubala R (2004) Vaccination with Leishmania soluble antigen and immunostimulatory oligodeoxynucleotides induces specific immunity and protection against Leishmania donovani infection. FEMS Immunol Med Microbiol 42(2):241–248

    Article  PubMed  CAS  Google Scholar 

  • Venkataprasad N, Coombes AGA, Singh M, Rohde M, Wilkinson K, Hudecz F, Davis SS, Vordermeier HM (1999) Induction of cellular immunity to a mycobacterial antigen adsorbed on lamellar particles of lactide polymers. Vaccine 17(15–16):1814–1819

    Article  PubMed  CAS  Google Scholar 

  • Verthelyi D, Kenney RT, Seder RA, Gam AA, Friedag B, Klinman DM (2002) Cpg oligodeoxynucleotides as vaccine adjuvants in primates. J Immunol (1950) 168(4):1659–1663

    CAS  Google Scholar 

  • Waterborg JH (2002) Quantitation of proteins. In: Walker J (ed) The protein protocols handbook, 2nd edn. Humana, New Jersey, pp 3–36

    Google Scholar 

  • Wu W, Weigand L, Belkaid Y, Mendez S (2006) Immunomodulatory effects associated with a live vaccine against Leishmania major containing CPG oligodeoxynucleotides. Eur J Immunol 36(12):3238–3247

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

This study was part of a research project which was financially supported by the vice chancellor for research of Mashhad University of Medical Sciences. We acknowledge the excellent technical assistance of Afshin Samiei and Dina Soroush.

Author information

Authors and Affiliations

  1. Nanotechnology and Pharmaceutical Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

    Mohsen Tafaghodi

  2. Biotechnology and Nanotechnology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

    Mahmoud R. Jaafari

  3. School of Pharmacy, Mashhad University of Medical Sciences, P.O. Box 91775-1365, Mashhad, Iran

    Mohsen Tafaghodi & Mahmoud R. Jaafari

  4. Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran, Iran

    Ali Khamesipour

Authors
  1. Mohsen Tafaghodi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ali Khamesipour
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mahmoud R. Jaafari
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Mahmoud R. Jaafari.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tafaghodi, M., Khamesipour, A. & Jaafari, M.R. Immunization against leishmaniasis by PLGA nanospheres encapsulated with autoclaved Leishmania major (ALM) and CpG-ODN. Parasitol Res 108, 1265–1273 (2011). https://doi.org/10.1007/s00436-010-2176-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00436-010-2176-4

Keywords

Search

Navigation