Early Ontogeny of Thymocytes in Pigs: Sequential Colonization of the Thymus by T Cell Progenitors

Šinkora, Marek; Sinkora, J; Rehakova, Zuzana; Butler, John E.

doi:10.4049/jimmunol.165.4.1832

Cited by 68 publications

(120 citation statements)

References 35 publications

(44 reference statements)

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“…Thymic gd T cells are characterized by the occurrence of CD1 + immature cells (13) that are always TCRgd med . Finding that CD1 + gd thymocytes are composed exclusively of CD2 + CD8 2 and CD2 + CD8 2 subsets agrees with previous work showing that the earliest precursors are CD1 + CD2 + CD8 2 gd T cells whereas CD1 + CD2 + CD8 + gd T cells are their progeny (5,11).…”

Section: Discussionmentioning

confidence: 99%

“…5A), resembling the periphery. However, thymic gd T cells contain developing precursors that can be identified by expression of CD1 (5,11,13). Whereas CD1 + gd thymocytes are strictly TCRgd med , CD1 2 gd thymocytes can be TCRgd med or TCRgd hi (Fig.…”

Section: Thymic Gd T Cells Can Be Divided Into Three Subsets Accordinmentioning

confidence: 99%

“…Thus, separation of CDR3 regions for TCRd on polyacrylamide sequencing gels provides a clonotypic analysis of porcine gd T cells showing their level of diversification (19). This procedure for measuring CDR3 polymorphism is called spectratyping and was performed essentially as described previously (5,12,13,19). Briefly, 50,000 or 100,000 sorted cells were immediately after sort dissolved in 0.5 ml TRI Reagent (Sigma-Aldrich).…”

Section: Pcr Amplification and Cdr3 Spectratypingmentioning

confidence: 99%

“…It is generally accepted that unlike ab T cells, gd T lymphocytes develop in the thymus without any selection for preantigen receptors (such as pre-TCRab) and also without any CD3 lo or TCR lo transitional stage (5,11,13,14). However, it is known that TCR can be expressed on gd T cells in two densities: medium and high (15,16).…”

mentioning

confidence: 99%

“…Our previous studies revealed the basic distribution and phenotypic profile of porcine gd T cells (3,12,18), their ontogeny (3,11), their development in the thymus (5,11,13), and the repertoire diversification of their TCR (19). In this study, we used high-speed flow cytometry sorting and cultivation techniques to characterize different subpopulation of gd T cells defined by TCRgd, CD2, and CD8 expression.…”

mentioning

confidence: 99%

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Porcine γδ T Lymphocytes Can Be Categorized into Two Functionally and Developmentally Distinct Subsets according to Expression of CD2 and Level of TCR

Štěpánová

Šinkora

2013

The Journal of Immunology

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Porcine γδ T cells have two levels of TCRγδ expression. Whereas TCRγδmed cells are mostly CD2+CD8− and CD2+CD8+, TCRγδhi cells are highly enriched for CD2−CD8−. This distribution is independent of bacterial colonization and it is already established in the thymus prior to export of γδ cells to the periphery. Sorting and cultivation experiments revealed that CD2−CD8− γδ cells are unable to acquire CD2 and CD8, whereas CD2+ subsets can gain or loose CD8. There is also differential susceptibility for proliferation between CD2+ and CD2− γδ cells. Although CD2−CD8− almost do not proliferate, proliferation of CD2+CD8− and CD2+CD8+ is substantial. Population of CD2− γδ cells is also absent in CD1+ immature thymocytes. Additionally, subpopulations of CD2+ and CD2− γδ cells in the thymus differ in expression of auxiliary surface molecules such as CD25, CD45RA/RC, and MHC class II. Moreover, TCRγδhi cells can generate TCRγδmed cells but never the opposite. The only exception is the thymus, where a few TCRγδmed cells can be induced to TCRγδhi but only under IL-2 influence. The repertoire of TCRδ is polyclonal in all subsets, indicating that there is the same extent of diversification and equal capability of immune responses. Results collectively indicate that CD2 expression determines two lineages of γδ cells that differ in many aspects. Because CD2− γδ cells are missing in the blood of humans and mice but are obvious in other members of γδ-high species such as ruminants and birds, our findings support the idea that circulating CD2− γδ T cells are a specific lineage.

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Section: Discussionmentioning

confidence: 99%

Section: Thymic Gd T Cells Can Be Divided Into Three Subsets Accordinmentioning

confidence: 99%

Section: Pcr Amplification and Cdr3 Spectratypingmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Porcine γδ T Lymphocytes Can Be Categorized into Two Functionally and Developmentally Distinct Subsets according to Expression of CD2 and Level of TCR

Štěpánová

Šinkora

2013

The Journal of Immunology

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Prenatal development of the porcine TCR δ repertoire: dominant expression of an invariant T cell receptor Vδ3‐Jδ3 chain

et al. 2004

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The prenatal development of the porcine + /ˇTCR repertoire was studied by complementarity-determining region 3 (CDR3) spectratyping and sequencing of TRDV1-DV5 transcripts. Specimens from the small and large intestine, spleen, thymus, liver, bone marrow and PBMC from fetal piglets between 38 and 114 days of gestation (DG) were examined. The TCRˇrepertoire was highly restricted early in gestation (DG38-DG57) and an invariant TRDV3 transcript, lacking the N/D region, was found in different fetuses throughout gestation and dominated the TRDV3 repertoires of all organs at mid gestation (˚DG55). Near the end of gestation, this invariant TRDV3 transcript was absent from the thymus but was still present, in a less dominant manner, in the intestine and spleen. The average CDR3 length of all Vˇsubgroups increased with ontogeny, suggesting an increase in activity of TdT. Thus, the persistence of fetal + /ˇT cells expressing an invariant TRDV3 chain throughout development is especially surprising since TdT is active early in gestation in swine. We speculate that these + /ˇT cells might have been selectively expanded by (self)-ligands and may have an important function throughout fetal development. IntroductionThe swine is one of the most important large-animal models belonging to " + /ˇhigh" species. They have an abundance of + /ˇT lymphocytes in their peripheral lymphoid pool [1][2][3][4], which makes them an attractive and useful model for the study of + /ˇT cells. However, only limited molecular data are available about porcine + /ˇT cells. Previously we have characterized the development and compartmentalization of the porcine TCRˇreper-toire at mucosal and extraintestinal sites after birth [5]. We have also demonstrated that the TCRˇrepertoire is highly polyclonal in young pigs and becomes increasingly oligoclonal and compartmentalized with age. These findings resemble those in humans [6] and non-human primates [7, 8]. In this study we characterized the prenatal development of the porcine TCRˇrepertoire and compared it with published data from human [6, 9, 10] and murine fetuses [11][12][13][14][15].Flow cytometric studies showed that + /ˇT cells are the earliest detectable T cell population in swine, developing first in the thymus around day of gestation (DG)40 and subsequently populating the periphery at about DG45 [2,[16][17][18]. This is also the case for mice [19], chicken [20,21] and humans [10,22,23] but not for sheep [24] and rats [25]. The proportion of + /ˇT cells decreases later in ontogeny (DG50-55) as § / g T cells expand and become the predominant CD3 + lymphocyte subset. This results in a predominance of § / g T cells in the second half of gestation in both the thymus and periphery [2,16,18]. Until DG90, peripheral + /ˇT cells account for only 3-5% of all lymphocytes [2,16]. However, near the end of gestation (DG90-114), there is a rapid expansion of + /ˇT cells in the periphery [2,16], so that + /ˇT cells in newborn and young piglets account for about 20% of all peripheral lymphocytes [2]. The prop...

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In utero transplanted human hepatocytes allow postnatal engraftment of human hepatocytes in pigs

et al. 2013

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In utero cell transplantation (IUCT) can lead to postnatal engraftment of human cells in the xenogeneic recipient. Most reports of IUCTs have involved hematopoietic stem cells. It is unknown if human hepatocytes used for IUCT in fetal pigs will lead to engraftment of these same cells in the postnatal environment. In this study, fetal pigs received direct liver injections of 1×107 human hepatocytes in utero and were delivered by cesarean-section at term. Piglets received a second direct liver injection of 5×107 human hepatocytes 1 week postnatally. Serum was analyzed for human albumin at 2, 4, and 6 weeks post-engraftment. Piglet livers were harvested 6 weeks after transplantation and examined by immunohistochemistry, PCR and fluorescence in situ hybridization for human specific sequences. Piglets receiving IUCT with human hepatocytes that were postnatally engrafted with human hepatocytes showed significant levels of human albumin production in their serum at all post-engraftment time points. Human albumin gene expression, the presence of human hepatocytes and the presence of human beta-2 microglobulin were all confirmed 6 weeks post-engraftment. IUCT in fetal pigs using human hepatocytes early in gestation allowed for engraftment of human hepatocytes, which remained viable and functional for weeks after transplantation. IUCT followed by postnatal engraftment may provide a future means for large scale expansion of human hepatocytes in genetically-engineered pigs.

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Early Ontogeny of Thymocytes in Pigs: Sequential Colonization of the Thymus by T Cell Progenitors

Cited by 68 publications

References 35 publications

Porcine γδ T Lymphocytes Can Be Categorized into Two Functionally and Developmentally Distinct Subsets according to Expression of CD2 and Level of TCR

Porcine γδ T Lymphocytes Can Be Categorized into Two Functionally and Developmentally Distinct Subsets according to Expression of CD2 and Level of TCR

Prenatal development of the porcine TCR δ repertoire: dominant expression of an invariant T cell receptor Vδ3‐Jδ3 chain

In utero transplanted human hepatocytes allow postnatal engraftment of human hepatocytes in pigs

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