Prenatal ontogeny of lymphocyte subpopulations in pigs

Šinkora, Marek; Sinkora, J; Rehakova, Zuzana; Šplı́chal, Igor; Yang, Huan; Parkhouse, R. M. E.; Trebichavsk, I

doi:10.1046/j.1365-2567.1998.00641.x

Cited by 63 publications

(114 citation statements)

References 26 publications

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“…Animals used in the study were Minnesota miniature/Vietnam-AsianMalaysian crossbred piglets bred in Novy Hradek (3,20). All pigs were healthy and normal at slaughter.…”

Section: Experimental Animalsmentioning

confidence: 99%

“…Cell suspensions were prepared essentially as previously described (3,21). Briefly, heparinized (20 U/ml; LECIVA-Zentiva) blood was obtained by intracardial puncture.…”

Section: Preparation Of Cell Suspensionsmentioning

confidence: 99%

“…The following mouse anti-pig mAbs, whose source and specificity were described earlier (3,4,21), were used as primary immunoreagents: antiTCRgd (PPT26, IgG1; PPT16, IgG2b; or PGBL22A, IgG1), anti-CD1 (76-7-4, IgG2a), anti-CD2 (MSA4, IgG2a; or 1038H-5-37, IgM), anti-CD8 (76-2-11, IgG2a; or PT36B, IgG1; VMRD, Pullman, WA), anti-CD25 (K231-3B2, IgG1), anti-CD45RA (FG2F9, IgG1), anti-CD45RC (MIL5, IgG1), and anti-swine MHC class II (MHC-II) leukocyte Ag type (SLA-DR; 1038H-12-34, IgM). Goat polyclonal Abs (pAbs) specific for mouse Ig subclasses labeled with FITC, PE, PE/Cy5 (alternatively PE/Cy7), or allophycocyanin were used as secondary immunoreagents (SouthernBiotech, Birmingham, AL).…”

Section: Immunoreagentsmentioning

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%

“…Traditionally, gd T cells in swine are subdivided into three subsets based on their expression of CD2 and CD8 and include CD2 2 CD8 2 , CD2 + CD8 2 , and CD2 + CD8 + cells (3)(4)(5)(6)(7). These individual subsets differ in their homing characteristic (8) and cytotoxic activities (7,9).…”

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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“…Animals used in the study were Minnesota miniature/Vietnam-AsianMalaysian crossbred piglets bred in Novy Hradek (3,20). All pigs were healthy and normal at slaughter.…”

Section: Experimental Animalsmentioning

confidence: 99%

“…Cell suspensions were prepared essentially as previously described (3,21). Briefly, heparinized (20 U/ml; LECIVA-Zentiva) blood was obtained by intracardial puncture.…”

Section: Preparation Of Cell Suspensionsmentioning

confidence: 99%

Section: Immunoreagentsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

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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