Étude typo-chronologique et stylistique des boucles d'oreilles en or de Macédoine : de l'époque archaïque à la fin de la période hellénistique

Jusqu’à présent, il n’existe aucun répertoire dédié exclusivement aux boucles d’oreilles macédoniennes et encore moins à celles uniquement en or. Ce mémoire veut remédier à cette pénurie en recensant, sous forme de catalogue, les boucles d'oreilles macédoniennes en or et en dégageant les tendances stylistiques de ces dernières à travers les périodes archaïque, classique et hellénistique. L’élaboration d’un catalogue aura été possible grâce à l’expertise de Vokotopoulou (Sindos), Ninou, Jackson, Despoinē, Grammenos, Kypraiou, Amandry et Descamps-Lequime sur les bijoux antiques. Leur analyse typologique et leur souci d’intégrer des exemplaires jusqu’alors non publiés auront certainement inspiré la forme de ce catalogue. Il conviendra d’abord d’y présenter une section portant sur les orfèvres et leur atelier d'orfèvrerie, les présentant comme des artisans mobiles dont le travail est généralement contractuel. Ces orfèvres pouvaient être indépendants de leur clientèle, tels des travailleurs autonomes, mais pouvaient à l’occasion être engagés par l’élite macédonienne pour confectionner des bijoux plus personnalisés et donc plus cher. Ensuite, nous présenterons un volet sur les techniques d’orfèvrerie afin de comprendre les termes techniques du domaine et apprécier adéquatement les niveaux de détails des boucles d’oreilles. Parmi les 147 paires de boucles d’oreilles recensées, nous avons déterminé 23 types dont 32 variantes se trouvent à travers 10 de ces types. Les types les plus dominants s'avèrent être les boucles d’oreilles «lion», les «bandes macédoniennes», les boucles d'oreilles «oméga» et les boucles d’oreilles «Éros». Ce contenu fut soumis à des analyses morphologiques, stylistiques, technologiques et même iconographiques pour certains exemplaires. Dans certains cas, il fut difficile de localiser le lieu exact de découverte en Macédoine alors que dans d’autres cas, le peu d’exemplaires d’un même type rend difficile l’interprétation de la représentation et son évolution stylistique.Until now, there are no exclusive repertoires of Macedonian earrings, especially those crafted out of gold. Thus, this thesis is meant to be the first to list the golden Macedonian earrings and their stylistic tendencies that emerged through the archaic, classical and hellenistic periods. The main inventories of jewellery established by Vokotopoulou (Sindos), Ninou, Jackson, Despoinē, Grammenos, Kypraiou, Amandry and Descamps-Lequime have certainly made this study possible due to their expertise on antique jewelry and by their concern to integrate unpublished artefacts. Inspired by these publications, a catalog based on typological analyses was established. Firstly, a section on goldsmiths and their workshops presents them as wandering craftsmen whose work is generally contractual. These goldsmiths could be at the same time independent of their clientele, such as self-employed and temporarily serving higher social castes to craft expensive and personalized jewellery. Finally, an extensive chapter on ancient goldsmithing techniques was unavoidable so that the reader can understand the technical terms of the domain and adequately visualize the levels of detail of the earrings. Out of the 147 pairs of earrings that have been identified, there are 23 distinct types of which 32 variants are found in 10 of these types. The most dominant types are lion earrings, Macedonian bands, omega earrings and Eros earrings. This content was subjected to morphological, stylistic, technological and even iconographic analyses for some. Certain of the attested types were confronted to an origin problem in Macedonia while others revealed an eminent lack of examples making both representation and stylistic evolution interpretation difficult

A nonintegrative lentiviral vector-based vaccine provides long-term sterile protection against malaria.

Trials testing the RTS,S candidate malaria vaccine and radiation-attenuated sporozoites (RAS) have shown that protective immunity against malaria can be induced and that an effective vaccine is not out of reach. However, longer-term protection and higher protection rates are required to eradicate malaria from the endemic regions. It implies that there is still a need to explore new vaccine strategies. Lentiviral vectors are very potent at inducing strong immunological memory. However their integrative status challenges their safety profile. Eliminating the integration step obviates the risk of insertional oncogenesis. Providing they confer sterile immunity, nonintegrative lentiviral vectors (NILV) hold promise as mass pediatric vaccine by meeting high safety standards. In this study, we have assessed the protective efficacy of NILV against malaria in a robust pre-clinical model. Mice were immunized with NILV encoding Plasmodium yoelii Circumsporozoite Protein (Py CSP) and challenged with sporozoites one month later. In two independent protective efficacy studies, 50% (37.5-62.5) of the animals were fully protected (p = 0.0072 and p = 0.0008 respectively when compared to naive mice). The remaining mice with detectable parasitized red blood cells exhibited a prolonged patency and reduced parasitemia. Moreover, protection was long-lasting with 42.8% sterile protection six months after the last immunization (p = 0.0042). Post-challenge CD8+ T cells to CSP, in contrast to anti-CSP antibodies, were associated with protection (r = -0.6615 and p = 0.0004 between the frequency of IFN-g secreting specific T cells in spleen and parasitemia). However, while NILV and RAS immunizations elicited comparable immunity to CSP, only RAS conferred 100% of sterile protection. Given that a better protection can be anticipated from a multi-antigen vaccine and an optimized vector design, NILV appear as a promising malaria vaccine

The protective efficacy of NILV was confirmed in a second independent study.

<p>Groups of BALB/c mice (n = 8/group) were immunized 3 times with NILV by intraperitoneal injections (in red) or not (in black). They were challenged with 500 spz injected IV one month after the last immunization. The % of parasitized red blood cells was followed every other day from day 5 to day 16 post-challenge by Giemsa-stained blood smears. Individual parasitemia are shown (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g007" target="_blank"><b>Figure 7A</b></a>) as well as means + SD (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g007" target="_blank"><b>Figure 7B</b></a>) and parasitemia at day 10 post-challenge (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g007" target="_blank"><b>Figure 7C</b></a>). Among the NILV-immunized mice (red circles), fully (•) <i>versus</i> partially (○) protected animals were further distinguished (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g007" target="_blank"><b>Figure 7D</b></a>). The Mann-Whitney test was used to compare NILV and naive and the Kruskal-Wallis test followed by a Dunn’s post-test were used to compare fully, partially and naive. The gross morphology of spleens and livers from NILV-immunized and naive mice at necropsy were compared 3 weeks post-challenge (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g007" target="_blank"><b>Figure 7E</b></a>).</p

NILV immunizations elicit enduring protective memory responses against malaria.

<p>Groups of BALB/c mice were immunized 3 times with NILV by intraperitoneal injections (n = 7) (in red) or not (n = 9) (in black). They were challenged with 500 spz injected IV six months after the last immunization (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g008" target="_blank"><b>Figure 8A</b></a>). The % of parasitized red blood cells was followed by Giemsa-stained blood smears. Individual parasitemia are shown (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g008" target="_blank"><b>Figure 8B</b></a>) as well as means + SD (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g008" target="_blank"><b>Figure 8C</b></a>) and parasitemia at day 9 post-challenge (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g008" target="_blank"><b>Figure 8D</b></a>). Among the NILV-immunized mice (in red), fully (•) <i>versus</i> partially (○) protected animals were further distinguished (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g008" target="_blank"><b>Figure 8E</b></a>).</p

Nonintegrative lentiviral vector encoding <i>Plasmodium yoelii</i> CSP used in the study.

<p>Lentiviral vector particles were produced by transient transfection of 293 T cells. The three plasmids used to generate particles are represented here (schematic representation not to scale). The vector expression plasmid pTRIP encodes the vaccine antigen, <i>Plasmodium yoelii</i> CSP. The encapsidation plasmid, p8.74 or pD64V for ILV or NILV respectively, codes for HIV-1 proteins required for particle formation and transduction. The envelope expression plasmid encodes non-crossreacting glycoproteins from Vesiculoviruses used in a specific order to circumvent anti-vector particles antibodies generated after each immunization (Vesicular Stomatitis Virus glycoprotein (VSV-G) Indiana (IND) serotype followed by VSV-G New Jersey (NJ) serotype followed by Cocal virus glycoprotein). Genes coding for structural/enzymatic and regulatory HIV-1 proteins are in dark and light blue respectively, while HIV-1 <i>cis</i>-acting sequences are in yellow and promoter sequences are in grey. The transferred gene, <i>Py</i> CSP, with a human codon-optimized sequence is in green. LTR, long terminal repeat; Ψ, encapsidation signal; cPPT/CTS, central polypurine tract/central termination sequence responsible for the formation of the DNA Flap structure during reverse-transcription which is a determinant of HIV-1 nuclear import; WPRE, Woodchuck hepatitis virus post-transcriptional response element to enhance mRNA nuclear export on a Rev/RRE independent fashion.</p

NILV are as immunogenic as ILV when 10-times more particles are injected.

<p>BALB/c mice (n = 5/group) were immunized by IM injection with various doses (expressed as TU/mouse) of lentiviral vector particles encoding Py CSP, either NILV (□) or ILV (▪). Ten days later, specific cellular immune responses were assessed (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g002" target="_blank"><b>Figure 2A</b></a>). The frequency of S9I-specific blood CD8+ cells was assessed by S9I/K^d tetramer staining, and the frequency of IFNg secreting splenocytes in response to overnight restimulation with S9I or I10L peptides was measured by IFNg elispot assay (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g002" target="_blank"><b>Figure 2B</b></a>). Means + SD are shown. BALB/c mice (n = 3/group) were IM immunized with NILV (□) or ILV (▪) at the dose of 5E+08 or 5E+07 TU/mouse respectively (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g002" target="_blank"><b>Figure 2C</b></a>). The frequency of S9I-specific blood CD8+ cells was followed over time by tetramer staining (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g002" target="_blank"><b>Figure 2D</b></a>). At day 24 post-immunization, spleen cellular response was analyzed by S9I/K^d tetramer staining (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g002" target="_blank"><b>Figure 2E</b></a>), by IFNg elispot in response to S9I and I10L peptides (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g002" target="_blank"><b>Figure 2F</b></a>), and by intracellular staining of 3 cytokines, IFNg, IL2 and TNFa, in response to S9I (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g002" target="_blank"><b>Figure 2G</b></a>). Cells secreting individual (green), 2 (blue) or 3 (red) cytokines are shown. Anti-(QGPGAP)₂-specific IgG at day 21 post-immunization were quantified by ELISA and expressed as titers (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g002" target="_blank"><b>Figure 2H</b></a>). Medians + range are shown.</p

NILV elicit as frequent blood CSP-specific T cells as RAS after 3 injections.

<p>BALB/c mice (n = 6/group) were immunized 3 times by IP injections of NILV. They were primed by administration of NILV particles encoding Py CSP and pseudotyped with VSV-G IND at the dose of 100 or 1500 ng p24/mouse. They were boosted 2 months later with 1500 ng p24 of NILV particles pseudotyped with VSV-G NJ, and boosted again 5 months later with 1500 ng p24 of NILV particles pseudotyped with the glycoprotein from Cocal virus. Additionally, mice (n = 6) from the same batch were immunized 3 times by IV injection with RAS at monthly intervals (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g003" target="_blank"><b>Figure 3A</b></a>). The frequency of S9I-specific blood CD8+ cells was followed over time by S9I/K^d tetramer staining after NILV (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g003" target="_blank"><b>Figure 3B</b></a>) and RAS immunizations (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g003" target="_blank"><b>Figure 3C</b></a>). Data from individual mice and means are shown. The Y-axis uses a logarithmic scale.</p

Protection is associated with CSP-specific CD8+ T cells responses.

<p>Immune correlates of protection against malaria were studied by plotting day 28 post-challenge immunity and day 9 post-challenge parasitemia as X and Y variables and using the Spearman test (the r_s and p values are shown) and linear regression (r² is shown) (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g006" target="_blank"><b>Figure 6A</b></a>). Immune responses in challenged mice were compared 28 days post-challenged between the vaccine candidates and their level of protection (fully (•) or partially (○) protected NILV immunized animals in red) by S9I/K^d tetramer staining and IFNg elispot assay with splenocytes and liver cells and elisa (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g006" target="_blank"><b>Figure 6B</b></a>). Means and SD are shown. The Kruskal-Wallis test was used to compare 3 or 4 groups, followed by a Dunn’s post-test.</p

Three immunizations with NILV and RAS induce comparable pre-challenge CSP-specific immune responses.

<p>Groups of BALB/c mice (n = 6/group) were immunized 3 times with NILV by IP injections (in red) or with RAS by IV injections (in black). Immune responses, both cellular and humoral, were compared 28 days after the last immunization. The frequency of IFNg secreting splenocytes in response to restimulation with S9I, I10L or S16I peptides was measured by IFNg elispot assay (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g004" target="_blank"><b>Figure 4A</b></a>). The quality of the S9I specific response was further studied by intracellular staining of 3 cytokines, IFNg, IL2 and TNFa, in response to S9I (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g004" target="_blank"><b>Figure 4B</b></a>). The frequency of IFNg secreting liver cells after restimulation with S9I was analyzed by IFNg elispot (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g004" target="_blank"><b>Figure 4C</b></a>). Additional mice (n = 6/group) were immunized to compare the vaccine-induced <i>in vivo</i> killing capacity of S9I-pulsed target cells (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g004" target="_blank"><b>Figure 4D</b></a>). The presence of IgG directed against (QGPGAP)₂ was assessed by ELISA (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048644#pone-0048644-g004" target="_blank"><b>Figure 4E</b></a>). Individual responses, means and SD are shown.</p