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  1. Article ; Online: Short-term culture adaptation of Toxoplasma gondii archetypal II and III field isolates affects cystogenic capabilities and modifies virulence in mice.

    Colos-Arango, Alicia / Largo-de la Torre, Andrea / Calero-Bernal, Rafael / Ortega-Mora, Luis-Miguel / Regidor-Cerrillo, Javier

    International journal for parasitology

    2023  Volume 53, Issue 9, Page(s) 491–504

    Abstract: Most Toxoplasma gondii research has been carried out using strains maintained in the laboratory for long periods of time. Long-term passage in mice or cell culture influences T. gondii phenotypic traits such as the capability to produce oocysts in cats ... ...

    Abstract Most Toxoplasma gondii research has been carried out using strains maintained in the laboratory for long periods of time. Long-term passage in mice or cell culture influences T. gondii phenotypic traits such as the capability to produce oocysts in cats and virulence in mice. In this work, we investigated the effect of cell culture adaptation in the short term for recently obtained type II (TgShSp1 (Genotype ToxoDB#3), TgShSp2 (#1), TgShSp3 (#3) and TgShSp16 (#3)) and type III (#2) isolates (TgShSp24 and TgPigSp1). With this purpose, spontaneous and alkaline stress-induced cyst formation in Vero cells during 40 passages, from passage 10 (p10) to 50 (p50), and isolate virulence at p10 versus p50 were studied using a harmonized bioassay method in Swiss/CD1 mice. T. gondii cell culture maintenance showed a drastic loss of spontaneous and induced production of mature cysts after ≈25-30 passages. The TgShSp1, TgShSp16 and TgShSp24 isolates failed to generate spontaneously formed mature cysts at p50. Limited cyst formation was associated with an increase in parasite growth and a shorter lytic cycle. In vitro maintenance also modified T. gondii virulence in mice at p50 with events of exacerbation, increasing cumulative morbidity for TgShSp2 and TgShSp3 isolates and mortality for TgShSp24 and TgPigSp1 isolates, or attenuation, with absence of mortality and severe clinical signs for TgShSp16, and better control of the infection with the lowest parasite and cyst burdens in lungs and brain for the TgShSp1 isolate. The present findings show deep changes in relevant phenotypic traits in laboratory-adapted T. gondii isolates and open new discussion about their use for inferring keys to parasite biology and virulence.
    MeSH term(s) Chlorocebus aethiops ; Animals ; Mice ; Cats ; Toxoplasma ; Toxoplasmosis, Animal/parasitology ; Virulence ; Vero Cells ; Genotype ; Antibodies, Protozoan
    Chemical Substances Antibodies, Protozoan
    Language English
    Publishing date 2023-04-12
    Publishing country England
    Document type Journal Article
    ZDB-ID 120518-3
    ISSN 1879-0135 ; 0020-7519
    ISSN (online) 1879-0135
    ISSN 0020-7519
    DOI 10.1016/j.ijpara.2023.03.004
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Short-term culture adaptation of Toxoplasma gondii archetypal II and III field isolates affects cystogenic capabilities and modifies virulence in mice

    Colos-Arango, Alicia / Largo-de la Torre, Andrea / Calero-Bernal, Rafael / Miguel Ortega-Mora, Luis / Regidor-Cerrillo, Javier

    International Journal for Parasitology. 2023 Apr. 12,

    2023  

    Abstract: Most Toxoplasma gondii research has been carried out using strains maintained in the laboratory for long periods of time. Long-term passage in mice or cell culture influences T. gondii phenotypic traits such as the capability to produce oocysts in cats ... ...

    Abstract Most Toxoplasma gondii research has been carried out using strains maintained in the laboratory for long periods of time. Long-term passage in mice or cell culture influences T. gondii phenotypic traits such as the capability to produce oocysts in cats and virulence in mice. In this work, we investigated the effect of cell culture adaptation in the short term for recently obtained type II (TgShSp1 (Genotype ToxoDB#3), TgShSp2 (#1), TgShSp3 (#3) and TgShSp16 (#3)) and type III (#2) isolates (TgShSp24 and TgPigSp1). With this purpose, spontaneous and alkaline stress-induced cyst formation in Vero cells during 40 passages, from passage 10 (p10) to 50 (p50), and isolate virulence at p10 versus p50 were studied using a harmonized bioassay method in Swiss/CD1 mice. Toxoplasma gondii cell culture maintenance showed a drastic loss of spontaneous and induced production of mature cysts after ≈ 25-30 passages. The TgShSp1, TgShSp16 and TgShSp24 isolates failed to generate spontaneously formed mature cysts at p50. Limited cyst formation was associated with an increase in parasite growth and a shorter lytic cycle. In vitro maintenance also modified T. gondii virulence in mice at p50 with events of exacerbation, increasing cumulative morbidity for TgShSp2 and TgShSp3 isolates and mortality for TgShSp24 and TgPigSp1 isolates, or attenuation, with absence of mortality and severe clinical signs for TgShSp16, and better control of the infection with the lowest parasite and cyst burdens in lungs and brain for the TgShSp1 isolate. The present findings show deep changes in relevant phenotypic traits in laboratory-adapted T. gondii isolates and open new discussion about their use for inferring keys to parasite biology and virulence.
    Keywords Toxoplasma gondii ; bioassays ; brain ; cell culture ; genotype ; morbidity ; mortality ; oocysts ; parasites ; parasitology ; phenotype ; virulence ; Type II and III genotypes ; Cell culture adaptation ; Phenotypic traits ; Cyst formation ; Mouse virulence
    Language English
    Dates of publication 2023-0412
    Publishing place Elsevier Ltd
    Document type Article ; Online
    Note Pre-press version ; Use and reproduction
    ZDB-ID 120518-3
    ISSN 1879-0135 ; 0020-7519
    ISSN (online) 1879-0135
    ISSN 0020-7519
    DOI 10.1016/j.ijpara.2023.03.004
    Database NAL-Catalogue (AGRICOLA)

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  3. Article ; Online: Archetypal type II and III <i>Toxoplasma gondii</i> oocysts induce different immune responses and clinical outcomes in experimentally infected piglets.

    Largo-de la Torre, Andrea / Diezma-Díaz, Carlos / Calero-Bernal, Rafael / Atencia-Cibreiro, Gabriela / Sánchez-Sánchez, Roberto / Ferre, Ignacio / Regidor-Cerrillo, Javier / Ortega-Mora, Luis Miguel

    Frontiers in immunology

    2022  Volume 13, Page(s) 1021556

    Abstract: Livestock animals, such as swine, are an important source of ... Toxoplasma gondii ... in the human population. Currently, there is limited knowledge regarding the potential influence that the ... T. gondii ... genotype might exert on establishing ... ...

    Abstract Livestock animals, such as swine, are an important source of Toxoplasma gondii in the human population. Currently, there is limited knowledge regarding the potential influence that the T. gondii genotype might exert on establishing infection in swine. Herein, we investigated the role of 2 T. gondii isolates, type II and III, representative of the genotypes circulating in Europe, in the immune responses and infection dynamics in piglets. Recently obtained oocysts (10<sup>3</sup>) from the T. gondii field isolates TgShSp1 (type II, ToxoDB genotype #3) and TgShSp24 (type III, #2) were used for oral infection. Thirteen 50-day-old female piglets of the Landrace-Large White crossbreed were randomly allocated into three different groups: Group 1 (G1, n=5), inoculated with TgShSp1; Group 2 (G2, n=5), inoculated with TgShSp24; and Group 3 (G3, n=3), a non-infected control group. Clinical signs were monitored daily until 42 days post-infection (dpi) when piglets were euthanized. Blood samples were collected weekly to test the cellular immune response in parasite-stimulated peripheral blood and specific IgG, IgG1 and IgG2, responses in sera. Parasite distribution and burden were evaluated in target tissues using a mouse bioassay and quantitative RT-PCR (qPCR). Apathy and a moderate decrease in feed consumption were observed in G1 and G2 piglets between 5 and 8 dpi, coinciding with fever (>40°C). G2 piglets had higher temperatures for a longer duration. Using mouse bioassay and qPCR, the detection frequency was higher in G2 vs. G1, and the highest parasite burdens in target tissues were also found in G2. Seroconversion was detected at 14 dpi in both infected groups, but higher antibody levels were observed in G2 piglets. Cytokine analyses revealed the production of IL-8, IL-1β and IFN-ɤ from 7 dpi in both infected groups. Moreover, IL-12 was produced from 7 dpi in G1 and from 14 dpi in G2. Levels of IL-8 were higher in G2, but IL-1β, IL-12 and IFN-ɤ were higher in G1 at 14 dpi. This cytokine profile reveals a predominant proinflammatory response that could be involved in limiting T. gondii infection in piglets, although it is more efficient against TgShSp1 type II-driven infection.
    MeSH term(s) Animals ; Female ; Immunity ; Immunoglobulin G ; Interleukin-12 ; Interleukin-8 ; Oocysts ; Swine ; Toxoplasma ; Toxoplasmosis, Animal
    Chemical Substances Immunoglobulin G ; Interleukin-12 (187348-17-0) ; Interleukin-8
    Language English
    Publishing date 2022-10-20
    Publishing country Switzerland
    Document type Journal Article ; Randomized Controlled Trial, Veterinary ; Research Support, Non-U.S. Gov't
    ZDB-ID 2606827-8
    ISSN 1664-3224 ; 1664-3224
    ISSN (online) 1664-3224
    ISSN 1664-3224
    DOI 10.3389/fimmu.2022.1021556
    Database MEDical Literature Analysis and Retrieval System OnLINE

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