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CD3-ε Overexpressed in Prothymocytes Acts as an Oncogene

Abstract

Background

Upon engagement of the T cell receptor for antigen, its associated CD3 proteins recruit signal transduction molecules, which in turn regulate T lymphocyte proliferation, apoptosis, and thymocyte development. Because some signal transducing molecules recruited by CD3-ε, i.e., p56lck and p59fyn, are oncogenic and since we previously found that overexpression of CD3-ε transgenes causes a block in T lymphocyte and NK cell development, we tested the hypothesis that aberrant CD3-ε signaling leads both to abnormal T lymphocyte death and lymphomagenesis.

Materials and Methods

Ten independently derived transgenic mouse lines were generated with four different genomic CD3-ε constructs. Mice either homozygous or hemizygous for each transgene were analyzed for an arrest in T lymphocyte development and for the occurrence of T cell lymphomas.

Results

Aggressive clonal T cell lymphomas developed at very high frequencies in seven mouse lines with intermediate levels of copies of CD3-ε derived transgenes. However, these lymphomas were not found when high copy numbers of CD3-ε transgenes caused a complete block in early thymic development or when a transgene was used in which the exons coding for the CD3-ε protein were deleted. Analyses of a series of double mutant mice, tgCD3-ε × RAG-2null, indicated that lymphomagenesis was initiated in lineage-committed prothymocytes, i.e., before rearrangement of the T cell receptor genes. In addition, the transgene coding for the CD3-ε cytoplasmic domain and its transmembrane region induced a T cell differentiation signal in premalignant tgCD3-ε × RAG-2null mice.

Conclusion

The nonenzymatic CD3-ε protein acted as a potent oncogene when overexpressed early in T lymphocyte development. Lymphomagenesis was dependent on signal transduction events initiated by the cytoplasmic domain of CD3-ε.

Introduction

Tumorigenesis involves perturbation of multiple signal transduction cascades resulting in a profound disturbance of normal control of cell cycling, cell growth, and/or cell death (13). Whereas mutated oncogenes and tumor suppressor genes play a pivotal role in these processes (49), adapters of signal transduction pathways can also be tumorigenic (1015). To investigate whether a surface receptor-associated protein that functions primarily by recruiting signal transduction molecules could also be tumorigenic, we analyzed large numbers of independently derived CD3-ε transgenic mice for spontaneous development of T cell tumors. CD3-ε is a T cell receptor (TCR)-associated membrane protein that plays a role in TCR/CD3 complex assembly and signal transduction (1620). Upon engagement of the T cell receptor with antigen, CD3-ε directed signal transduction pathways are initiated that regulate T cell proliferation, apoptosis, and thymocyte development (2125). Because some signal transducing molecules recruited by CD3-ε, i.e., p56lck and; p59fyn, are oncogenic (26,27), a disturbance of CD3-ε signaling could potentially lead to abnormal T lymphocyte death and lymphomagenesis.

As previously reported in a number of independently derived homozygous CD3-ε transgenic mice (i.e., tgε26+/+), a very early block in thymocyte and natural killer (NK) cell development is observed. This block in development is caused by overexpression of the CD3-ε proteins derived from the transgenes that are regulated by their own cis-regulatory elements (28,29). Here we demonstrate that in hemizygous transgenic mouse lines, overexpression of CD3-ε resulted in an extremely high incidence of very aggressive T cell lymphomas. The lymphomas were observed in seven mouse lines carrying relatively high copy numbers of transgenes encoding CD3-ε-derived proteins with the capacity to recruit signal transduction pathways. However, lymphomas were not observed in transgenic lines in which CD3-ε expression was low or absent. These data indicate that a small nonenzymatic protein with a 55 amino acid cytoplasmic tail is an oncogene that can induce lymphomas in T lymphocyte precursors.

Materials and Methods

Mice

Generation and screening of transgenic mice carrying all DNA constructs were carried out as described previously (28,29). For reasons of simplicity, we renamed the constructs in this report: pL12 as ε, pL12Δ1 as εΔl, pL12Δ2 as εΔ2, and pL16 as εm (see ref. 28,29). RAG-2null mice were obtained from GenPharm International (Palo Alto, CA). lcknull mice were kindly provided by Dr. T.W. Mak. Double mutant RAG-2null × tgε26, RAG-2null × tgεΔl, and lcknull(ref. 30) × tg826 mice were obtained by breeding. The mice were housed in virus antibody-free (V.A.F.) conditions at the Animal Research Facility of Beth Israel Deaconess Medical Center.

Detection of Thymic Lymphomas

The frequency of lymphomas is a summary of mice with thymomas when sacrificed at 8 months of age and mice that died of lymphomas during that 8-month period, divided by the total number of mice monitored (40 or more unless otherwise stated). The frequency is an underestimation because the thymomas were scored by visual inspection of the thymuses and not by pathologic examination.

Flow Cytometry

Flow cytometric analysis of thymocytes and tumor cells for surface antigen expression was performed by three-color analysis as described previously (29).

Antibody Treatment of RAG-2null mice

Antibody treatment of RAG-2null mice was performed as described previously (31). Briefly, young RAG-2null mice (8 to 20 days after birth) were injected i.p. with 10 µg of anti-CD3ε mAb per gram body weight, and sacrificed for analysis 4 weeks later.

Histology

Tissues were frozen at −20°C and mounted for cryostat sectioning. Sections 5 µm thick were fixed in 4% paraformaldehyde, phosphate-buffered saline (PBS) (pH 7.4). The sections were stained with hematoxylin and eosin, or used for immunohistochemistry as described below. The sections were blocked with 2% normal rat serum, PBS (pH 7.4) for 20 min, stained with biotinylated primary antibodies for 1 hr, followed by ABC reagent (Vector Labs, Burlington, CA) for 45 min. The sections were counterstained with hematoxylin, dehydrated, and mounted. Biotinylated rat anti-mouse Thy-1.2 antibody was purchased from PharMingen (San Diego, CA).

Results

High Incidence of T cell Lymphomas in Hemizygous tgε26 mice

When 40 homozygous and hemizygous tgε26 mice were monitored over a period of 1 year, almost all hemizygous tgε26+/− mice died of T cell lymphomas during that time period. The earliest tumor-induced death occurred at 3 months of age and the lymphomas resulted in 85% mortality by 8 months in these animals (Fig. 1, Table 1). In contrast, no tumors were detectable in homozygous tgε26+/+ mice (Fig. 1), which is consistent with the fact that tgε26+/+ mice did not develop any T lymphocytes. The lymphomas observed in tgε26+/− mice were always found first in the thymus, with the enlargement of one lobe visible from 8 weeks of age and older. In the later stages of the disease, the enlarged thymuses were up to 20 times their respective normal sizes, and enlarged spleens and lymph nodes were also frequently observed (Fig. 2A). Histological studies of these animals revealed metastases of the lymphomas in the trachea, lungs, liver, kidney, testis, and brain (Fig. 2B, and data not shown).

Fig. 1
figure 1

Survival curve for tgε26+/+ and tgε26+/− mice.

Forty tgε26+/+ and 40 tgε26+/− mice were monitored for lymphoma-induced death. The data illustrate the high incidence of T cell lymphomas in tgε26+/− mice.

Fig. 2
figure 2

Visual and histological examination of lymphomas in tgε26+/− mice.

(A) Comparison of the thymus, lymph nodes, and spleen from a tgε26+/− mouse at a late stage of lymphoma with the counterpart organs from a wild-type mouse. (B) Immunohistology of tumor metastasis to nonlymphoid organs. Thy-1 stained tissue sections from (a) trachea (25×), (b) liver (160×), and (c) kidney (160×) of a hemizygous tgε26 mouse in a late stage of the disease.

Table 1 Frequency of T cell lymphomas in CD3-ε transgenic mice

Flow cytometric analyses revealed that the thymic lymphomas were of T cell origin representing different stages of thymocyte development, the majority being CD4+8+ (Table 2). Southern blotting with a TCR-Cβ probe indicated that the tumors were generally clonal, as 28 of 39 tumors (72%) had one or two rearranged TCR-β bands (data not shown, see Discussion). Upon transfer of these tumor cells into immunodeficient animals, i.e., nude, RAG-2null, and homozygous tgε26 mice, or in syngeneic wild-type mice, death occurred within 3 to 4 weeks. Stable cell lines could be derived from approximately of the tumors by in vitro tissue culture of the tumor cells without added growth factors. Generally, these cell lines resembled their parental tumors phenotypically (data not shown). Together, these observations support the notion that CD3-ε transgenes were involved in the generation of T lymphomas that were initiated early in T lymphocyte development.

Table 2 Frequency of major T cell lymphoma phenotypes in transgenic mice

T cell Lymphomas Are Caused by Transgenic CD3-ε Protein

To exclude the possibility that the T cell lymphomas in tgε26+/− mice were induced by an insertional mutation of the transgene, nine additional CD3-ε transgenic mouse lines (hemizygous and homozygous) were monitored for eight months (Table 1). These mice were generated with three different CD3-ε genomic constructs: transgenes εΔ1 and εΔ2 representing the human CD3-ε gene with two different deletions, and transgene εm, representing a chimeric human-murine CD3-ε gene, coding for the murine CD3-ε protein (28,29). During a period of 8 months, high frequencies of T cell lymphomas were found in six of the nine lines. Timing of onset and phenotypes of these T lymphomas were similar to those in tgε26+/− (Tables 1 and 2). Thus, a position effect of the CD3-ε transgene in tumorigenesis was ruled out. Moreover, these data indicated that either the human or murine CD3-ε protein could be oncogenic.

All seven lymphoma-prone transgenic lines had two important features in common: they carried relatively high copy numbers of different transgenes (>20×), and they were partially, but not completely, T cell deficient (Table 1). The frequency of tumors observed in several groups of hemizygous transgenic mice (e.g., tgεΔ1 or tgεm) increased with an increase in the number of transgene copies in those lines (Table 1). Importantly, in the same group of mice, the level of CD3-ε protein expression in thymocytes also increased approximately with the increase in the number of the transgene copies (29). We conclude, therefore, that this transgene-induced lymphomagenesis was dependent upon overexpression of the CD3-ε protein. This notion was strongly supported by the absence of tumors in animals with a wild-type phenotype owing to low transgene copy numbers (e.g., tgεΔ1-2966L) and in animals with high numbers of copies of transgene εΔ2 which did not contain the majority of CD3-ε coding sequences (Table 1).

Whereas no tumors were found in homozygous mice with a complete block in T cell development e.g., tgε26+/+ and tgεΔ1-2978+/+, T cell tumors were found in homozygous mice with partial T cell development (Table 1). For example, the thymic cellularity of the homozygous tgεΔ1-2966H mice was on average 13% of that in wild-type litter mates, and 50% of these tgεΔ1-2966H+/+ mice developed T lymphomas in 8 months (Table 1). The frequency of tumors in the homozygous tgεΔ1-2966H mice was even higher than that of the hemizygous mice of the same strain (50% versus 15%; see Table 1). Since the levels of transgene expression in homozygous mice were higher than those in hemizygous mice (29, and data not shown), this observation also supported the notion that overexpression of CD3-ε was involved in tumorigenesis.

As shown in Table 1, high frequencies of lymphomas were observed in tgεΔ1-2978+/− and 2966H mice. In the εΔ1 construct, the region encoding the ectodomain of CD3-ε was deleted from the original human genomic fragment, resulting in a truncated CD3-ε polypeptide that consisted of the 55 amino acid cytoplasmic domain and its membrane-anchor. This result suggests that the oncogenic signal could be initiated by this small nonenzymatic polypeptide.

CD3-ε-induced Tumorigenesis Occurs in Lineage-committed Prothymocytes

CD3-ε-induced tumor cells isolated from the thymus often represented T lymphocytes that were more immature than tumor cells derived from the spleen or lymph nodes of the same mice (Fig. 3, and data not shown). This finding, together with the observation that lymphomas were always first detected in the thymuses of tumor-bearing animals, indicate that cell transformation occurred in thymocytes. Since our previous experiments demonstrated that overexpression of the CD3-ε-derived transgenes began at or prior to Day 13 of gestation (29), i.e., in prothymocytes, we examined whether the lymphomas were initiated in prothymocytes. To this end, two of the CD3-ε transgenic lines were bred with RAG-2null mice that have a block in T cell development at the CD44CD25+CD4CD8 control point (32). Indeed, as shown in Table 3, T cell lymphomas were frequently found in RAG-2null × tgε26+/−and RAG-2null × tgεΔ1-2966H+/− mice, but not in mice that were RAG-2null × tgε26+/+ or RAG-2null without the transgene. These results indicate that the CD3-ε-induced tumorigenesis occurred prior to the rearrangement and expression of the T cell receptor.

Fig. 3
figure 3

Two-color fluorescent profiles of lymphocytes isolated from thymuses and lymph nodes of 5 tgε26+/ mice.

These data show the differentiation of lymphocytes after cell transformation.

Table 3 Expression of CD3-ε-derived transgenes induces transition from DN to DP thymocytes in young RAG-2null and lcknull mice and lymphomagenesis in older mice

All tgε/tgεΔ1 × RAG-2null lymphoma cells represented immature thymocytes, as they were surface CD3 (data not shown). Some tumors had a phenotype of Thy-1+CD44CD25+CD4 CD8 (Fig. 4A, tumor #1, and data not shown), representing very immature thymocytes (33,34). Interestingly, most tumors from the double mutant mice were CD4+CD8+CD3 (Fig. 4A, tumor #2, and data not shown), suggesting that the CD 3-ε transgene might have signaled by itself to cause a partial progression of the tumor cell past the CD44CD25CD4CD8 control point (32).

Fig. 4
figure 4

Phenotypes of thymic lymphoid tumor cells isolated from double mutant mice.

(A) Flow cytometric analyses of tumor cells derived from RAG-2null × tgε26+/− mice. The cells were stained with antibodies against CD4 and CD8, and CD25 and CD44. Most of the tumors from these mice bear the phenotype represented by tumor #2.

(B) Flow cytometric analyses of tumor cells derived from lcknull × tgε26+/− mice. The cells were stained with antibodies against CD3, CD4, and CD8.

Signal Transduction by CD3-ε Cytoplasmic Tail in Double-negative Thymocytes

Several investigators have shown that CD 3-ε could be triggered in CD48 (DN) RAGnull thymocytes by in vivo treatment with anti-CD3-ε antibodies (23,31,35). The anti-CD3-ε effect on DN thymocytes from RAG-1null or RAG-2null mice can be measured in terms of thymocyte proliferation, induction of the CD4 and CD8α and CD8β genes, as well as the abrogation of RAG-1 or RAG-2 transcription (23,36). To examine whether the transgene coding for the CD3-ε cytoplasmic tail could initiate signal transduction pathways in lineage-committed pretumor thymocytes, double mutant mice, i.e., RAG-2null × tgε26+/− and RAG-2null × tgεΔ1-2966H+/−, were analyzed. In young (4 to 6 weeks) RAG-2null × tgεΔ1-2966H+/− mice, the transgene induced a transition from DN to DP (CD4+8+) thymocytes, along with down-regulation of CD25, and a moderate increase in cellularity (Fig. 5, Table 3). In RAG-2null × tgε26+/− and RAG-2null × tgεΔ1-2966H+/+ mice, a similar transition of DN to DP thymocytes and the down-regulation of CD25 were observed (Table 3). In the latter mice, however, the thymocyte cellularity was lower than in the antibody-induced transition, most likely because of the apoptosis of early thymocytes (Table 3). These observations indicate that overexpression of the CD3-ε cytoplasmic domain in prethymocytes mimicked the effects of anti-CD3-ε activation.

Fig. 5
figure 5

Expression of CD3-ε derived transgenes induces transition from DN to DP thymocytes in young RAG-2null mice.

Thymocytes were analyzed by three-color staining with combinations of antibodies against Thy-1, CD44, and CD25; and CD4, CD8, and TCR-αβ. Thymocytes were derived from a young RAG-2null × tgεΔ1-2966H+/− mouse, from a RAG-2null mouse 1 month after i.p injection of an anti-CD3-ε antibody, and from a RAG-2null control mouse. Most of the thymocytes (>90%) from all of these mice were Thy-l+ and TCR-αβ (not shown).

Previously, we showed that in RAG-1null × lcknull mice, transition from DN to DP thymocytes can be only partially induced by anti-CD3 activation, which suggests that lck is an important element in signal transduction through the TCR/CD 3 complex during the early stages of T cell development (23). However, lcknull × tgε26+/− double mutant mice developed lymphomas at a high frequency (Table 3 and Fig. 4b). Taken together, these analyses on double mutant mice shown in Table 3 and Figures 4 and 5 demonstrate that the cytoplasmic domain of CD3-ε could signal in DN thymocytes and that lck was not the only protein kinase involved in this signaling.

Discussion

In this report we have shown that CD3-ε could function as an oncogene, since overexpression of the CD3-ε protein in prothymocytes was essential for the induction of T cell tumors. This conclusion is based on the following evidence: (1) tumors were observed in transgenic mice expressing functional CD3-ε-derived proteins, but not in transgenic mice made with a genomic construct that did not have any protein expression (8A2); (2) the frequency of tumor incidence increased with an increase in the copy number of transgenes; (3) the level of transgene expression in thymocytes and peripheral T cells of these mice also increased with the copy number of transgenes (29); and (4) transgenes were overex-pressed in immature thymocytes (from embryonic day 13 onwards) in mice carrying high copy numbers (29). By contrast, mice overexpressing human CD3-δ do not have an arrest in T lymphocyte development, nor do they develop lymphomas (29; B. Wang et al., unpublished data).

Overexpression of CD3-ε-induced cell transformation may be initiated in very immature thymocytes, for instance, in Thy-1+CD44+CD25 thymocytes, since high frequencies of T cell lymphomas were observed in double mutant RAG-2null × tgε26+/− and RAG-2null × tgεΔ1-2966H+/− mice. The transformed prothymocytes could continue to differentiate to give more mature phenotypes. Since tumors isolated from the transgenic mice (on RAG-2+/+ background) were generally clonal (one or two rearranged TCR-β genes on Southern blots), an unknown secondary event(s) may occur in the transformed. TCR-rearranged thymocytes, leading to the formation of clonal lymphomas. This notion is consistent with the theory of multistep tumorigenesis (2). One may predict that if the secondary event(s) occurs in prothymocytes prior to TCR gene rearrangement, lymphoma cells derived from one progenitor cell could continue to differentiate to display polyclonal TCR gene rearrangements. Indeed, no dominant rearranged TCR-β band(s) was detected in 28% of the lymphomas (11 of 39 examined) from the transgenic mice on a RAG-2+/+ background. It is noteworthy that timing and clonality of T cell lymphomagenesis in tgε26+/− mice were similar to those found in transgenic mice overexpressing lck, in transgenic mice expressing the chimeric homeobox gene E2A-PBX1 (37), and in hemizygous Ikaros mutant mice (38). However, there are some differences between these systems. We demonstrated that lck is not essential for CD3-ε induced T cell lymphomas, because both CD3+ and CD3 tumor cells were observed in the double mutant lcknull × tgε26+/− mice. In contrast, only CD3 tumors were found in the transgenic mice overexpressing lck (26). In hemizygous Ikaros mutant mice, T cell lymphomagenesis is preceded by a lymphoproliferation in thymocytes and peripheral T cells (38). This phenomenon was not observed in thymocytes and peripheral T cells derived from prelymphoma tgε26+/− mice.

We propose that overexpression of CD3-ε generates a signal to induce cell transformation, and we further postulate that the signal is related to normal signal transduction through CD3-ε. This hypothesis is supported by the observation that overexpression of CD3-ε mimics the effects induced by anti-CD3-ε treatment of thymocytes in vivo (23,31). For instance, overexpression of CD 3-ε in thymocytes prevents intracellular TCR-β expression, induces apoptosis (29; D. Zheng et al., unpublished observations), and induces a transition from DN to DP in RAG-2null mice. Moreover, this hypothesis is consistent with earlier reports suggesting that murine leukemia virus-induced leukemogenesis is mediated through a signal(s) triggered by the binding of the virus with T cell receptors (39,40). Signal transduction through CD 3-ε involves lck and other c-src-like kinases (20,41,42). Although lck is dispensable for CD3-ε-induced T cell lymphomas, it is still conceivable that induction of T cell tumorigenesis by CD 3-ε and lck share common downstream pathways. Importantly, unlike lck, the 55 amino acid cytoplasmic tail of CD3-ε is not a protein tyrosine kinase, nor does it have any known enzymatic activity. The role of CD3-ε in oncogenesis may therefore lie in its function as a provider of docking sites for kinases and other signal transduction enzymes. There may be some similarity with nonenzymatic adapter proteins. This notion is consistent with recent observations that She, one of the nonenzymatic adapter proteins involved in a variety of signal transduction pathways, can induce cell transformation in in vitro culture systems (1015). Nonenzymatic receptor proteins other than CD3-ε may therefore be potential oncogenes under some circumstances.

References

  1. Korsmeyer SJ. (1993) Programed cell death: BcI-2. Important Adv. Oncol. 93: 19–28.

    Google Scholar 

  2. Vogelstein B, Kinzler KW. (1993) The multistep nature of cancer. Trends Genet. 9: 138–141.

    Article  CAS  PubMed  Google Scholar 

  3. Boettiger D. (1989) Interaction of oncogenes with differentiation programs. Curr. Top. Microbiol. Immunol. 147: 31–78.

    PubMed  CAS  Google Scholar 

  4. Hall M, Peters G. (1996) Genetic alterations of cychns, cyclin-dependent kinases, and cdk inhibitors in human cancer. Adv. Cancer Res. 68: 67–109.

    Article  CAS  PubMed  Google Scholar 

  5. Seiivanova G, Wiman KG. (1995) p53: A cell cycle regulator activated by DNA damage. Adv. Cancer Res. 66: 143–180.

    Article  Google Scholar 

  6. Leonard CJ, Canman CE, Kastan MB. (1995) The role of p53 in cell-cycle control and apoptosis: Implications for cancer. Important Adv. Oncol. 95: 33–42.

    Google Scholar 

  7. Wang JY, Knudsen ES, Welch PJ. (1994) The retinoblastoma tumor suppressor protein. Adv. Cancer Res. 64: 25–85.

    Article  CAS  PubMed  Google Scholar 

  8. Fisher RJ, Bader JP, Papas TS. (1989) Oncogenes and the mitogenic signal pathway. Important Adv. Oncol. 89: 3–27.

    Google Scholar 

  9. Balmain A, Brown K. (1988) Oncogene activation in chemical carcinogenesis. Adv. Cancer Res. 51: 147–182.

    Article  CAS  PubMed  Google Scholar 

  10. Dilworth SM, Brewster CE, Jones MD, Lanfrancone L, Pelicci G, Pelicci PG. (1994) Transformation by polyoma virus middle Tantigen involves the binding and tyrosine phosphorylation of She. Nature 367: 87–90.

    Article  CAS  PubMed  Google Scholar 

  11. Crowe AJ, McGlade J, Pawson T, Hayman MJ. (1994) Phosphorylation of the SHC proteins on tyrosine correlates with the transformation of fibroblasts and erythroblasts by the v-sea tyrosine kinase. Oncogene 9: 537–544.

    PubMed  CAS  Google Scholar 

  12. Harrison-Pindik D, Susa M, Varticovski L. (1995) Association of phosphatidylinositol 3-kinase with SHC in chronic myelogeneous leukemia cells. Oncogene 10: 1385–1391.

    Google Scholar 

  13. Pelicci G, Lanfrancone L, Grignani F, McGlade J, Cavallo F, Forni G, Nicoletti I, Grignani F, Pawson T, Pelicci PG. (1992) A novel transforming protein (SHC) with an SH2 domain is implicated in mitogenic signal transduction. Cell 70: 93–104.

    Article  CAS  PubMed  Google Scholar 

  14. Pelicci G, Lanfrancone L, Salcini AE, Romano A, Mele S, Grazia Borrello M, Segatto O, Di Fiore PP, Pelicci PG. (1995) Constitutive phosphorylation of Shc proteins in human tumors. Oncogene 11: 899–907.

    PubMed  CAS  Google Scholar 

  15. Salcini AE, McGlade J, Pelicci G, Nicoletti I, Pawson T, Pelicci PG. (1994) Formation of Shc-Grb2 complexes is necessary to induce neoplastic transformation by overexpression of Shc proteins. Oncogene 9: 2827–2836.

    PubMed  CAS  Google Scholar 

  16. Clevers H, Alarcon B, Wilwman T, Terhorst C. (1988) The T cell receptor/CD3 complex: A dynamic protein ensembe. Annu. Rev. Immunol. 6: 629–662.

    Article  CAS  PubMed  Google Scholar 

  17. Ashwell JD, Klusner RD. (1990) Genetic and mutational analysis of the T-cell antigen receptor. Annu. Rev. Immunol. 8: 139–167.

    Article  CAS  PubMed  Google Scholar 

  18. Letourneur F, Klausner RD. (1992) Activation of T cells by a tyrosine kinase activation domain in the cytoplasmic tail of CD3 ε. Science 255: 79–82.

    Article  CAS  PubMed  Google Scholar 

  19. Wegener A-MK, Letourneur F, Hoeveler A, Brocker T, Luton F, Malissen B. (1992) The T cell receptor/CD3 complex is composed of at least two autonomous transduction modules. Cell 68: 83–95.

    Article  CAS  PubMed  Google Scholar 

  20. Terhorst C, Regueiro JR. (1992) T cell activation. In: Lachmann PJ et al. (eds) Clinical Aspects of Immunology Blackwell Science, Inc. Oxford, UK. Vol 1, pp. 447–466.

    Google Scholar 

  21. Owen JJT, Owen MJ, Williams GT, Kingston R, Jenkinson EJ. (1988) The effects of anti-CD3 antibodies on the development T-cell receptor αβ lymphocytes in embryonic thymus organ cultures. Immunology 63: 639–642.

    PubMed  PubMed Central  CAS  Google Scholar 

  22. Bentin J, Vaughan JH, Tsoukas CD. (1988) T cell proliferation induced by anti-CD3 antibodies: Requirement for a T-T cell interaction. Eur. J. Immunol. 18: 627–632.

    Article  CAS  PubMed  Google Scholar 

  23. Levelt CN, Mombaerts P, Wang B, Kohler H, Tonegawa S, Eichmann K, Terhorst C. (1995) Regulation of thymocyte development through CD3: Functional dissociation between p56lck and CD3 sigma in early thymic selection. Immunity 3: 215–222.

    Article  CAS  PubMed  Google Scholar 

  24. Malissen M, Gillet A, Ardouin L, Bouvier G, Trucy J, Ferrier P, Vivier E, Malissen B. (1995) Altered T cell development in mice with a targeted mutation of the CD3-ε gene. EMBO J. 14: 4641–4653.

    PubMed  PubMed Central  CAS  Article  Google Scholar 

  25. Shinkai Y, Ma A, Cheng HL, Alt FW. (1995) CD3 epsilon and CD3 zeta cytoplasmic domains can independently generate signals for T cell development and function. Immunity 2: 401–411.

    Article  CAS  PubMed  Google Scholar 

  26. Abraham KM, Levin SD, Marth JD, Forbush KA, Perlmutter RM. (1991) Thymic tumori- genesis induced by overexpression of p56lck. Proc. Natl Acad. Sci. U.S.A. 88: 3977–3981.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Anderson SJ, Levin SD, Perlmutter RM. (1994) Involvement of the protein tyrosine kinase p56lck in T cell signaling and thymocyte development. Adv. Immunol. 56: 151–178.

    Article  CAS  PubMed  Google Scholar 

  28. Wang B, Biron C, She J, Higgins K, Sunshine MJ, Lacy E, Lonberg N, Terhorst C. (1994) A block in both early T lymphocyte and natural killer cell development in transgenic mice with high-copy numbers of the human CD3E gene. Proc. Natl. Acad. Sci. U.S.A. 91: 9402–9406.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Wang B, Levelt CN, Salio M, Zheng D-X, Sancho J, Liu C-P, She J, Huang M, Higgins K, Sunshine M-J, Eichmann K, Lacy L, Lonberg N, Terhorst C. (1995) Over-expression of CD36ε transgenes blocks T lymphocyte development. Int. Immunol. 7: 435–448.

    Article  CAS  PubMed  Google Scholar 

  30. Molina TJ, Kishihara K, Siderovski DP, et al. (1992) Profound block in thymocyte development in mice lacking p56lck. Nature 357: 161–164.

    Article  CAS  PubMed  Google Scholar 

  31. Levelt CN, Mombaerts P, Iglesias A, Tonegawa S, Eichmann K, (1993) Restoration of early thymocyte differentiation in T-cell receptor beta-chain-deficient mutant mice by transmembrane signaling through CD3 epsilon. Proc. Natl. Acad. Sci. U.S.A. 90: 11401–11405.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Shinkai Y, Rathbun G, Lam K-P, Oltz EM, Stewart V, Mendelsohn M, Charron J, Datta M, Young F, Stall AM, Alt FW. (1992) Rag-2-deficient mice lack mature lymphocytes owing to inability to initiate V(D)J rearrangement. Cell 68: 855–867.

    Article  CAS  PubMed  Google Scholar 

  33. Godfrey DI, Zlotnik A. (1993) Control points in early T-cell development. Immunol. Today 14: 547–553.

    Article  CAS  PubMed  Google Scholar 

  34. Scollay R, Wilson A, D’Amico A, Kelly K, Egerton M, Pearse M, Wu L, Shortman K. (1988) Developmental status and reconstitution potential of subpopulations of murine thymocytes. Immunol. Rev. 104: 81–120.

    Article  CAS  PubMed  Google Scholar 

  35. Shinkai Y, Alt FW. (1994) CD3 epsilon-mediated signals rescue the development of CD4+CD8+ thymocytes in RAG-2-/- mice in the absence of TCR beta chain expression. Int. Immunol. 6: 995–1001.

    Article  CAS  PubMed  Google Scholar 

  36. Levelt CN, Wang B, Ehrfeld A, Terhorst C, Eichmann K. (1995) Regulation of T cell receptor (TCR)-beta locus allelic exclusion and initiation of TCR-alpha locus rearrangement in immature thymocytes by signaling through the CD3 complex. Eur. J. Immunol. 25:1257–1261.

    Article  CAS  PubMed  Google Scholar 

  37. Dedera DA, Waller EK, LeBrun DP, Sen-Majumdar A, Stevens ME, Barsh GS, Cleary ML. (1993) Chimeric homeobox gene E2A-PBXI induces proliferation, apoptosis, and malignant lymphomas in transgenic mice. Cell 74: 833–843.

    Article  CAS  PubMed  Google Scholar 

  38. Winandy S, Wu P, Georgopoulos K. (1995) A dominant mutation in the Ikaros gene leads to rapid development of leukemia and lymphoma. Cell 83: 289–299.

    Article  CAS  PubMed  Google Scholar 

  39. O’Neill HC, McGrath MS, Allison JP, Weissman IL. (1987) A subset of T cell receptors associated with L3T4 molecules mediates C6VL leukemia cell binding of its cognate retrovirus. Cell 49: 143–151.

    Article  PubMed  Google Scholar 

  40. McGrath MS, Pillemer E, Weissman IL. (1980) Murine leukaemogenesis: Monoclonal antibodies to T-cell determinants arrest T-lymphoma cell proliferation. Nature 285: 259–261.

    Article  CAS  PubMed  Google Scholar 

  41. Anderson SJ, Perlmutter RM. (1995) A signaling pathway governing early thymocyte maturation. Immunol. Today 16: 99–105.

    Article  CAS  PubMed  Google Scholar 

  42. Weiss A, Littman DR. (1994) Signal transduction by lymphocyte antigen receptors. Cell 76: 263–274.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

We thank Key Higgins for technical assistance, and Drs. Clyde Dawe, Tom Benjamin and Geoffrey Cooper for a critical review of the manuscript. This work was supported by a grant from the National Institutes of Health (PO1-AI 35714).

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Wang, B., She, J., Salio, M. et al. CD3-ε Overexpressed in Prothymocytes Acts as an Oncogene. Mol Med 3, 72–81 (1997). https://doi.org/10.1007/BF03401669

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Keywords

  • Prothymocytes
  • Double Mutant Mice
  • Recruit Signal Transduction
  • Thymocyte Development
  • Lymphocyte Death