Limits...
The single positive T cells found in CD3-zeta/eta-/- mice overtly react with self-major histocompatibility complex molecules upon restoration of normal surface density of T cell receptor-CD3 complex.

Lin SY, Ardouin L, Gillet A, Malissen M, Malissen B - J. Exp. Med. (1997)

Bottom Line: CD3-zeta/eta-deficient mice have small thymuses containing cells that show a profound reduction in the surface levels of T cell receptors and terminate their differentiation at the CD4+CD8+ stage.Fusion of these peripheral T cells with a CD3-zeta-positive derivative of the BW5147 TCR-alpha-/beta- thymoma resulted in hybridomas that do express an heterogeneous set of T cell receptor alpha/beta dimers at their surface and at density comparable to those found in hybridomas derived from wild-type peripheral T cells.These results demonstrate that by increasing the density and/or output of the T cell receptors expressed in peripheral T cells, one can confer them with the capacity to respond to normal density of self-MHC molecules.

View Article: PubMed Central - PubMed

Affiliation: Centre d'Immunologie, Institut National de la Sante et de la Recherche Medicale-Centre National de Recherche Scientifique de Marseille-Luminy, France.

ABSTRACT
CD3-zeta/eta-deficient mice have small thymuses containing cells that show a profound reduction in the surface levels of T cell receptors and terminate their differentiation at the CD4+CD8+ stage. Rather unexpectedly, CD3- or very low single positive T cells accumulate over time in the spleen and lymph nodes of CD3-zeta/eta-deficient mice after a process dependent on MHC expression. Fusion of these peripheral T cells with a CD3-zeta-positive derivative of the BW5147 TCR-alpha-/beta- thymoma resulted in hybridomas that do express an heterogeneous set of T cell receptor alpha/beta dimers at their surface and at density comparable to those found in hybridomas derived from wild-type peripheral T cells. We have investigated the specificities of these T cell receptors using spleen cells from congenic and mutant mouse strains, and showed that the majority of them readily recognized self-MHC class I or class II molecules. These results demonstrate that by increasing the density and/or output of the T cell receptors expressed in peripheral T cells, one can confer them with the capacity to respond to normal density of self-MHC molecules.

Show MeSH

Related in: MedlinePlus

CD4+CD8− and CD4−CD8+ cells accumulate over time in  the lymph nodes of CD3-ζ/η–deficient mice. Cells from thymus and  lymph nodes were isolated from 1- to 3-mo-old CD3-ζ/η–deficient mice  and analyzed by two-color flow cytometry for the expression of CD4  versus CD8. The percentage of cells found in each quadrant is indicated.  Most of the lymph node cells scoring as CD4−CD8− were brightly  stained with an anti-B220 antibody (RA3-6B2), and likely belong to the  B cell lineage.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2196153&req=5

Figure 1: CD4+CD8− and CD4−CD8+ cells accumulate over time in the lymph nodes of CD3-ζ/η–deficient mice. Cells from thymus and lymph nodes were isolated from 1- to 3-mo-old CD3-ζ/η–deficient mice and analyzed by two-color flow cytometry for the expression of CD4 versus CD8. The percentage of cells found in each quadrant is indicated. Most of the lymph node cells scoring as CD4−CD8− were brightly stained with an anti-B220 antibody (RA3-6B2), and likely belong to the B cell lineage.

Mentions: When analyzed by flow cytometry for the expression of CD4 and CD8 molecules, thymuses from 1- to 3-mo-old CD3-ζ/η−/− mice were found to contain dramatically fewer single positive cells than age-matched wild-type thymuses (Fig. 1). Moreover, most of these cells expressed residual levels of CD25 molecules and probably corresponded to “immature” single positive cells on the way to becoming double positive cells (20). Despite the near total absence of mature single positive thymocytes, Thy1+-single positive T cells were found to accumulate over time in lymph nodes of CD3-ζ/η−/− mice (Fig. 1, right), but in smaller numbers than those from age-matched wild-type littermates (CD3-ζ/η+/+, n = 5; average 13 × 106; CD3-ζ/η−/−, n = 7, average 5.8 × 106). Similar to CD3-ζ/η−/− thymocytes, the single positive cells found in CD3-ζ/η−/− lymph nodes expressed levels of TCR/CD3 complexes barely detectable by flow cytometry (Fig. 2; 8–10, 17). When the lymph node T cell populations found in 3-mo-old wild-type and CD3-ζ/η−/− mice were compared for the expression of various cell surface markers, they both showed identical staining profiles for CD25, CD62, CD69, CD122, and Fas molecules as well as a NK1.1− phenotype (Fig. 2 and data not shown). The most striking difference was seen in the level of expression of CD44 which was highly expressed by all the single positive cells found in CD3-ζ/η−/− lymph nodes (Fig. 2). A corresponding population of Thy1+, CD3− or very low single positive T cells was also present in the spleen of 3-mo-old CD3-ζ/η–deficient mice (data not shown). It should be noted that we have not been able to extend our analysis of CD3-ζ/η−/− over a longer time because mice die around 4 mo of age after developing a chronic intestinal inflammation, characterized by diarrhea and wasting, similar to that previously documented in cytokine or TCR mutant mice (27).


The single positive T cells found in CD3-zeta/eta-/- mice overtly react with self-major histocompatibility complex molecules upon restoration of normal surface density of T cell receptor-CD3 complex.

Lin SY, Ardouin L, Gillet A, Malissen M, Malissen B - J. Exp. Med. (1997)

CD4+CD8− and CD4−CD8+ cells accumulate over time in  the lymph nodes of CD3-ζ/η–deficient mice. Cells from thymus and  lymph nodes were isolated from 1- to 3-mo-old CD3-ζ/η–deficient mice  and analyzed by two-color flow cytometry for the expression of CD4  versus CD8. The percentage of cells found in each quadrant is indicated.  Most of the lymph node cells scoring as CD4−CD8− were brightly  stained with an anti-B220 antibody (RA3-6B2), and likely belong to the  B cell lineage.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196153&req=5

Figure 1: CD4+CD8− and CD4−CD8+ cells accumulate over time in the lymph nodes of CD3-ζ/η–deficient mice. Cells from thymus and lymph nodes were isolated from 1- to 3-mo-old CD3-ζ/η–deficient mice and analyzed by two-color flow cytometry for the expression of CD4 versus CD8. The percentage of cells found in each quadrant is indicated. Most of the lymph node cells scoring as CD4−CD8− were brightly stained with an anti-B220 antibody (RA3-6B2), and likely belong to the B cell lineage.
Mentions: When analyzed by flow cytometry for the expression of CD4 and CD8 molecules, thymuses from 1- to 3-mo-old CD3-ζ/η−/− mice were found to contain dramatically fewer single positive cells than age-matched wild-type thymuses (Fig. 1). Moreover, most of these cells expressed residual levels of CD25 molecules and probably corresponded to “immature” single positive cells on the way to becoming double positive cells (20). Despite the near total absence of mature single positive thymocytes, Thy1+-single positive T cells were found to accumulate over time in lymph nodes of CD3-ζ/η−/− mice (Fig. 1, right), but in smaller numbers than those from age-matched wild-type littermates (CD3-ζ/η+/+, n = 5; average 13 × 106; CD3-ζ/η−/−, n = 7, average 5.8 × 106). Similar to CD3-ζ/η−/− thymocytes, the single positive cells found in CD3-ζ/η−/− lymph nodes expressed levels of TCR/CD3 complexes barely detectable by flow cytometry (Fig. 2; 8–10, 17). When the lymph node T cell populations found in 3-mo-old wild-type and CD3-ζ/η−/− mice were compared for the expression of various cell surface markers, they both showed identical staining profiles for CD25, CD62, CD69, CD122, and Fas molecules as well as a NK1.1− phenotype (Fig. 2 and data not shown). The most striking difference was seen in the level of expression of CD44 which was highly expressed by all the single positive cells found in CD3-ζ/η−/− lymph nodes (Fig. 2). A corresponding population of Thy1+, CD3− or very low single positive T cells was also present in the spleen of 3-mo-old CD3-ζ/η–deficient mice (data not shown). It should be noted that we have not been able to extend our analysis of CD3-ζ/η−/− over a longer time because mice die around 4 mo of age after developing a chronic intestinal inflammation, characterized by diarrhea and wasting, similar to that previously documented in cytokine or TCR mutant mice (27).

Bottom Line: CD3-zeta/eta-deficient mice have small thymuses containing cells that show a profound reduction in the surface levels of T cell receptors and terminate their differentiation at the CD4+CD8+ stage.Fusion of these peripheral T cells with a CD3-zeta-positive derivative of the BW5147 TCR-alpha-/beta- thymoma resulted in hybridomas that do express an heterogeneous set of T cell receptor alpha/beta dimers at their surface and at density comparable to those found in hybridomas derived from wild-type peripheral T cells.These results demonstrate that by increasing the density and/or output of the T cell receptors expressed in peripheral T cells, one can confer them with the capacity to respond to normal density of self-MHC molecules.

View Article: PubMed Central - PubMed

Affiliation: Centre d'Immunologie, Institut National de la Sante et de la Recherche Medicale-Centre National de Recherche Scientifique de Marseille-Luminy, France.

ABSTRACT
CD3-zeta/eta-deficient mice have small thymuses containing cells that show a profound reduction in the surface levels of T cell receptors and terminate their differentiation at the CD4+CD8+ stage. Rather unexpectedly, CD3- or very low single positive T cells accumulate over time in the spleen and lymph nodes of CD3-zeta/eta-deficient mice after a process dependent on MHC expression. Fusion of these peripheral T cells with a CD3-zeta-positive derivative of the BW5147 TCR-alpha-/beta- thymoma resulted in hybridomas that do express an heterogeneous set of T cell receptor alpha/beta dimers at their surface and at density comparable to those found in hybridomas derived from wild-type peripheral T cells. We have investigated the specificities of these T cell receptors using spleen cells from congenic and mutant mouse strains, and showed that the majority of them readily recognized self-MHC class I or class II molecules. These results demonstrate that by increasing the density and/or output of the T cell receptors expressed in peripheral T cells, one can confer them with the capacity to respond to normal density of self-MHC molecules.

Show MeSH
Related in: MedlinePlus