Membrane Proteins and Enzymes of Leishmania

Schneider, Pascal; Bordier, Clément; Etges, Robert

doi:10.1007/978-1-4899-1651-8_2

Cited by 13 publications

(10 citation statements)

References 151 publications

(114 reference statements)

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“…These enzymes are encoded by the lmcpb gene family (24,25,38,39). The second was a zinc metalloproteinase, a glycoprotein of 63 kDa (GP63) carrying a glycosylphosphatidylinositol (GPI) membrane anchor, which is the major surface protein of promastigotes (0.5 to 1% of the cellular protein corresponding to about 5 ϫ 10 5 molecules/cell); this protein is present in promastigotes of all Leishmania species investigated (53). In contrast, L. mexicana amastigotes from mouse lesion tissue express only very small amounts of GP63 on their surface (32).…”

Section: Resultsmentioning

confidence: 99%

Subunit Vaccination of Mice against New World Cutaneous Leishmaniasis: Comparison of Three Proteins Expressed in Amastigotes and Six Adjuvants

et al. 2000

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A mixture of well-defined recombinant antigens together with an adjuvant that preferentially stimulates specific gamma interferon (IFN-␥)-secreting helper type 1 CD4 ؉ T cells (Th1 cells) presents a rational option for a vaccine against leishmaniasis. The potential of this approach was investigated in murine infections with Leishmania mexicana, which are characterized by the absence of a parasite-specific Th1 response and uncontrolled parasite proliferation. A mixture of three antigens (glycoprotein 63, cysteine proteinases, and a membrane-bound acid phosphatase), which are all expressed in amastigotes, the mammalian stage of the parasite, were used for the immunization of C57BL/6 mice in combination with six adjuvants (interleukin 12 [IL-12], Detox, 4-monophosphoryl lipid A, QS-21, Mycobacterium bovis BCG, and Corynebacterium parvum). All six vaccine formulations containing the mixture of recombinant antigens were protective against challenge infections with promastigotes, the insect stage of the parasite, in that mice controlled and healed infections but developed transient and, in certain cases, accentuated disease. The most effective adjuvants were IL-12 followed by Detox. Further studies using these two adjuvants showed that a similar protective effect was observed with a mixture of the corresponding native proteins, and mice which had controlled the infection showed a preponderance of IFN-␥-secreting CD4 ؉ T cells in the lymph nodes draining the lesion. Using the recombinant proteins individually, it is shown that the relatively abundant cysteine proteinases and glycoprotein 63, but not the acid phosphatase, are able to elicit a protective response. The results are discussed in comparison to previous studies with subunit vaccines and with respect to cell biological aspects of antigen presentation in Leishmania-infected macrophages.The solid immunity observed following recovery from cutaneous leishmaniasis in humans has stimulated numerous attempts for the development of a prophylactic vaccination against this widespread (sub)tropical disease (12,16,19,35,36). Leishmania major, the etiological agent of Old World cutaneous leishmaniasis, has been the most popular species both in studies of murine infections and in human trials. In the Middle East, the deliberate infection with L. major was a common and effective practice for immunization against subsequent infections, but a fraction of the vaccinated persons produced lesions that required medical treatment. A vaccine based on killed promastigotes, the insect stage of the parasite, and Mycobacterium bovis BCG was recently shown to be ineffective in a controlled trial with several thousand volunteers in Iran (37). Mice have been used in a range of vaccination protocols against infection by L. major. Representative examples include: attenuated but live parasites (23,29,34,48,59); subunit vaccines delivered by live carriers such as BCG expressing the surface proteinase GP63 of L. major (11); vaccinia virus expressing the glycoprotein GP46/M-2 (20, 30); GP63 expr...

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Section: Resultsmentioning

confidence: 99%

Subunit Vaccination of Mice against New World Cutaneous Leishmaniasis: Comparison of Three Proteins Expressed in Amastigotes and Six Adjuvants

et al. 2000

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“…Membrane-active fungicidal peptides, devoid of bactericidal activity, have been described [15]. NO production and apoptosis in the monocytic cell line Raw is induced by the cecropin Amelittin hybrid peptide CA(1-8)M (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18), but not by other shorter analogues with similar bactericidal activity [16].…”

Section: Introductionmentioning

confidence: 99%

“…Leishmania is a protozoan parasite that affects humans as well as other mammalian hosts. The insect form of the parasite is the Abbreviations used : BCECF-AM, 2h,7h-bis-(2-carboxyethyl)-5(6)-carboxyfluorescein triacetoxymethyl ester ; Bisoxonol, bis-(1,3-diethylthiobarbituric) trimethine oxonol ; Boc, t-butyloxycarbonyl ; CA(1-8)M (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18), KWKLFKKIGIGAVLKVLTTGLPALIS-NH 2 ; FCCP, carbonyl cyanide p-trifluoromethoxyphenylhydrazone ; LPG, lipophosphoglycan ; MALDI-TOF, matrix-assisted laser desorption/ionization time-of-flight spectroscopy ; MTT, 3-(4,5dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide ; NMGCl, N-methylglucamide chloride ; PG, phosphoglycan. 1 To whom correspondence should be addressed.…”

Section: Introductionmentioning

confidence: 99%

The plasma membrane of Leishmania donovani promastigotes is the main target for CA(1–8)M(1–18), a synthetic cecropin A–melittin hybrid peptide

Díaz‐Achirica

Ubach

Guinea

et al. 1998

Biochemical Journal

102

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Reports on the lethal activity of animal antibiotic peptides have largely focused on bacterial rather than eukaryotic targets. In these, involvement of internal organelles as well as mechanisms different from those of prokaryotic cells have been described. CA(1-8)M(1-18) is a synthetic cecropin A-melittin hybrid peptide with leishmanicidal activity. Using Leishmania donovani promastigotes as a model system we have studied the mechanism of action of CA(1-8)M(1-18), its two parental peptides and two analogues. At micromolar concentration CA(1-8)M(1-18) induces a fast permeability to H+/OH-, collapse of membrane potential and morphological damage to the plasma membrane. Effects on other organelles are related to the loss of internal homeostasis of the parasite rather than to a direct effect of the peptide. Despite the fast kinetics of the process, the parasite is able to deactivate in part the effect of the peptide, as shown by the higher activity of the d-enantiomer of CA(1-8)M(1-18). Electrostatic interaction between the peptide and the promastigote membrane, the first event in the lethal sequence, is inhibited by polyanionic polysaccharides, including its own lipophosphoglycan. Thus, in common with bacteria, the action of CA(1-8)M(1-18) on Leishmania promastigotes has the same plasma membrane as target, but is unique in that different peptides show patterns of activity that resemble those observed on eukaryotic cells.

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“…The striking modifications concerned the parasite surface protease Gp63 (Medina-Acosta et al 1989;Frommel et al 1990) (Sacks & Da Silva, 1987;Sacks, Brodin & Turco, 1990). Moreover, studies on the basic cellular function of these organisms led to the identification of several membrane proteins and enzymes which are essential for the parasite survival in its hosts (Schneider, Bordier & Etges, 1993). Some of these membrane enzymes play a role in the degradation of nutrients which could be taken as simplified components from the external milieu.…”

Section: Introductionmentioning

confidence: 99%

Characterization of aLeishmaniaantigen associated with cytoplasmic vesicles resembling endosomal-like structure

et al. 1993

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In the present study we have used antibodies to Leishmania major promastigote antigens which were eluted from a glutathione-agarose column (LmGbp) and could identify several parasite components among different Leishmania species by using immunoprecipitation and Western blot techniques. The results also showed that some of LmGbp are present among the molecules released into the culture medium. Moreover, immunofluorescence assays clearly demonstrated that LmGbp are expressed by intracellular amastigotes. The electron micrographs of thawed cryosections of L. major-infected cells revealed that the antigens were associated with the membrane of the phagocytic vacuole. Moreover, the Western blot technique allowed us to identify, using other Leishmania species extracts and anti-LmGbp antibodies, a major polypeptide of an apparent molecular mass of 66 kDa. Immunofluorescence studies suggested that the 66 kDa polypeptide is associated with intracytoplasmic vesicles. Cryosections of Leishmania promastigotes improved the fine structure preservation of the organelles and enabled a number of features to be seen, particularly the structures considered as vesicles, which appeared as a complex tubulo-vesicular structure resembling mammalian cell endosomes and Leishmania organelles previously named 'megasomes'. Further studies using antibodies against the native 66 kDa protein will be needed to investigate the localization of the protein at the ultrastructural level and to follow its intracellular vesicular traffic.

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Membrane Proteins and Enzymes of Leishmania

Cited by 13 publications

References 151 publications

Subunit Vaccination of Mice against New World Cutaneous Leishmaniasis: Comparison of Three Proteins Expressed in Amastigotes and Six Adjuvants

Subunit Vaccination of Mice against New World Cutaneous Leishmaniasis: Comparison of Three Proteins Expressed in Amastigotes and Six Adjuvants

The plasma membrane of Leishmania donovani promastigotes is the main target for CA(1–8)M(1–18), a synthetic cecropin A–melittin hybrid peptide

Characterization of aLeishmaniaantigen associated with cytoplasmic vesicles resembling endosomal-like structure

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