Expression Cloning of a Protective
            <i>Leishmania</i>
            Antigen

Mougneau, Evelyne; Altare, Frédéric; Wakil, Adil E.; Zheng, Shichun; Coppola, Thierry; Wang, Zhi-En; Waldmann, Rainer; Locksley, Richard M.; Glaichenhaus, Nicholas

doi:10.1126/science.7725103

Cited by 333 publications

(306 citation statements)

References 33 publications

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“…Upon adoptive transfer into naive recipients, LC loaded with a mixture of the recombinant Leishmania Ag LACK, KMP-11, gp63, and PSA were shown to mediate significant protection against a challenge with L. major parasites. The significance of LC as the vaccine carrier is emphasized by the previous observation that gp63, LACK, and KMP-11 are unable to induce protective responses when administered as proteins without adjuvant (23,28,31). Most importantly, we identified LeIF as a single leishmanial Ag that, upon loading into LC, protected mice from uncontrolled parasite infection.…”

Section: Discussionmentioning

confidence: 85%

Dendritic Cell (DC)-Based Protection Against an Intracellular Pathogen Is Dependent Upon DC-Derived IL-12 and Can Be Induced by Molecularly Defined Antigens

Berberich

Ramírez-Pineda

Hambrecht

et al. 2003

The Journal of Immunology

110

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Upon loading with microbial Ag and adoptive transfer, dendritic cells (DC) are able to induce immunity to infections. This offers encouragement for the development of DC-based vaccination strategies. However, the mechanisms underlying the adjuvant effect of DC are not fully understood, and there is a need to identify Ag with which to arm DC. In the present study, we analyzed the role of DC-derived IL-12 in the induction of resistance to Leishmania major, and we evaluated the protective efficacy of DC loaded with individual Leishmania Ag. Using Ag-pulsed Langerhans cells (LC) from IL-12-deficient or wild-type mice for immunization of susceptible animals, we showed that the inability to release IL-12 completely abrogated the capacity of LC to mediate protection against leishmaniasis. This suggests that the availability of donor LC-derived IL-12 is a requirement for the development of protective immunity. In addition, we tested the protective effect of LC loaded with Leishmania homolog of receptor for activated C kinase, gp63, promastigote surface Ag, kinetoplastid membrane protein-11, or Leishmania homolog of eukaryotic ribosomal elongation and initiation factor 4a. The results show that mice vaccinated with LC that had been pulsed with selected molecularly defined parasite proteins are capable of controlling infection with L. major. Moreover, the protective potential of DC pulsed with a given Leishmania Ag correlated with the level of their IL-12 expression. Analysis of the cytokine profile of mice after DC-based vaccination revealed that protection was associated with a shift toward a Th1-type response. Together, these findings emphasize the critical role of IL-12 produced by the sensitizing DC and suggest that the development of a DC-based subunit vaccine is feasible.

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Section: Discussionmentioning

confidence: 85%

Dendritic Cell (DC)-Based Protection Against an Intracellular Pathogen Is Dependent Upon DC-Derived IL-12 and Can Be Induced by Molecularly Defined Antigens

Berberich

Ramírez-Pineda

Hambrecht

et al. 2003

The Journal of Immunology

110

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“…The use of a proven protective T cell line as the readout of the T cell expression cloning approach is an attractive alternative. This approach, initially developed for the identification of a leishmanial and a listerial gene encoding T cell Ags (45,46), was recently described as a powerful strategy for the direct screening and cloning of genes from a M. tuberculosis genomic expression library (36) using human M. tuberculosis reactive T cell lines as readouts. Here, the use of this technology, employing a proven protective murine CD4 ϩ T cell line, resulted in the cloning of several potentially protective genes.…”

Section: Discussionmentioning

confidence: 99%

T Cell Expression Cloning of aMycobacterium tuberculosisGene Encoding a Protective Antigen Associated with the Early Control of Infection

Skeiky

Ovendale

Jen

et al. 2000

The Journal of Immunology

112

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Infection of C57BL/6 mice with Mycobacterium tuberculosis results in the development of a progressive disease during the first 2 wk after challenge. Thereafter, the disease is controlled by the emergence of protective T cells. We have used this infection model in conjunction with direct T cell expression cloning to identify Ags involved with the early control of the disease. A protective M. tuberculosis-specific CD4 T cell line derived from mice at 3 wk postchallenge was used to directly screen an M. tuberculosis genomic expression library. This screen resulted in the identification of a genomic clone comprising two putative adjacent genes with predicted open reading frames of 10 and 41 kDa, MTB10 and MTB41, respectively (the products of Rv0916c and Rv0915c, respectively, in the TubercuList H37Rv database). MTB10 and MTB41 belong to the PE and PPE family of proteins recently identified to comprise 10% of the M. tuberculosis genome. Evaluation of the recombinant proteins revealed that MTB41, but not MTB10, is the Ag recognized by the cell line and by M. tuberculosis-sensitized human PBMC. Moreover, C57BL/6 mice immunized with MTB41 DNA developed both CD4- (predominantly Th1) and CD8-specific T cell responses to rMTB41 protein. More importantly, immunization of C57BL/6 mice with MTB41 DNA induced protection against infection with M. tuberculosis comparable to that induced by bacillus Calmette-Guérin. Thus, the use of a proven protective T cell line in conjunction with the T cell expression cloning approach resulted in the identification of a candidate Ag for a subunit vaccine against tuberculosis.

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“…This protein has been shown to be highly conserved among Leishmania species evaluated to date and has been shown to confer protection in susceptible mice when administered together with IL-12 [8]. However, the expansion of LACK-reactive CD4 1 T cells have been shown to be the focus of the early response in mouse strains susceptible to L. major infection [9].…”

Section: Introductionmentioning

confidence: 99%

Induction and abrogation of LACK reactive cells in the evolution of human leishmaniasis

Maasho

Wolday

Edjigu

et al. 2001

Clinical and Experimental Immunology

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SUMMARY Peripheral blood mononuclear cells (PBMC) from cutaneous leishmaniasis patients with ongoingLeishmania aethiopica infection and individuals cured/under treatment from L. infantum or L. donovani infection were stimulated in vitro with LACK, the Leishmania homologue of receptors for activated C kinase. The LACK protein is conserved in related leishmanial species and is expressed both in the promastigote and amastigote stages of Leishmania. Our results show that LACK induced marked NK and some CD8 1 cell proliferation in PBMC from cutaneous leishmaniasis patients with active disease. These responses were coupled with high levels of IFN-g and IL-10 production. At the concentration tested, the proliferative responses to freeze-thawed Leishmania antigen (Ft-Leish) were higher, while the levels of IFN-g were consistently lower than that of LACK. Although cells from individuals cured of leishmaniasis could respond to whole Leishmania lysate by proliferation and IFN-g production, there was no evident response to LACK. Ethiopian controls tested at the same time also showed LACK induced proliferation with IFN-g and IL-10 responses. Thus LACK reactivity in terms of proliferation and cytokine induction were present in cells from some healthy donors and most of the patients with active lesions, while this response was absent in individuals cured of L. infantum or L. donovani leishmaniasis. Since cure from leishmaniasis often results in life-long protection, and active but not cured patients showed in vitro responses to LACK stimulation, questions arose as to how this highly immunodominant molecule functions during human leishmanisasis. Some possible mechanisms are discussed.

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Expression Cloning of a Protective Leishmania Antigen

Cited by 333 publications

References 33 publications

Dendritic Cell (DC)-Based Protection Against an Intracellular Pathogen Is Dependent Upon DC-Derived IL-12 and Can Be Induced by Molecularly Defined Antigens

Dendritic Cell (DC)-Based Protection Against an Intracellular Pathogen Is Dependent Upon DC-Derived IL-12 and Can Be Induced by Molecularly Defined Antigens

T Cell Expression Cloning of aMycobacterium tuberculosisGene Encoding a Protective Antigen Associated with the Early Control of Infection

Induction and abrogation of LACK reactive cells in the evolution of human leishmaniasis

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