Alteration of early haematopoietic development is thought to be responsible for the onset of immature leukemias and lymphomas. We have previously demonstrated that Tax_HTLV-1 interferes with ß-selection, an important checkpoint of early thymopoiesis, indicating that human T-cell leukemia virus type 1 (HTLV-1) infection has the potential to perturb thymic human αβ T-cell development. To verify that inference and to clarify the impact of HTLV-1 infection on human T-cell development, we investigated the in vivo effects of HTLV-1 infection in a “Human Immune System” (HIS) Rag2^-/-γ_c^-/- mouse model. These mice were infected with HTLV-1, at a time when the three main subpopulations of human thymocytes have been detected. In all but two inoculated mice, the HTLV-1 provirus was found integrated in thymocytes; the proviral load increased with the length of the infection period. In the HTLV-1-infected mice we observed alterations in human T-cell development, the extent of which correlated with the proviral load. Thus, in the thymus of HTLV-1-infected HIS Rag2^-/-γc^-/- mice, mature single-positive (SP) CD4⁺ and CD8⁺ cells were most numerous, at the expense of immature and double-positive (DP) thymocytes. These SP cells also accumulated in the spleen. Human lymphocytes from thymus and spleen were activated, as shown by the expression of CD25: this activation was correlated with the presence of tax mRNA and with increased expression of NF-kB dependent genes such as bfl-1, an anti-apoptotic gene, in thymocytes. Finally, hepato-splenomegaly, lymphadenopathy and lymphoma/thymoma, in which Tax was detected, were observed in HTLV-1-infected mice, several months after HTLV-1 infection. These results demonstrate the potential of the HIS Rag2^-/-γ_c^-/- animal model to elucidate the initial steps of the leukemogenic process induced by HTLV-1. Human T-cell leukemia virus type 1 (HTLV-1) is a retrovirus responsible for the development of Adult T cell Leukemia, an aggressive malignant lymphoproliferation that occurs several decades after viral exposure in some individuals infected when neonates via breastfeeding. Here, we used mice that were humanized by transplantation of human hematopoietic stem cells isolated from cord blood. Such an animal model may reproduce key aspects of in vivo infection with HTLV-1 and improve the understanding of the early steps of the leukemogenic process. These humanized mice when inoculated with HTLV-1 were indeed productively infected, as shown by the detection and the expression of the provirus in human cells. We observed that T-cell development in the thymus of these mice was profoundly altered, as ascertained by the increased proliferation of activated mature T lymphocytes that accumulated in the thymus as well as in the spleen. These alterations of human thymic T-cell development may be linked to the development of ATL–like pathologies. These results show that this humanized mouse model provides an invaluable tool not only to study the early stages of the infection leading to the development of malignant lymphoproliferations, but also to design new therapeutic approaches.

Human T-cell leukemia virus type 1 (HTLV-1) is the etiological agent of adult T-cell leukemia/lymphoma (ATLL), an aggressive malignant proliferation of activated CD4+ T lymphocytes. The viral Tax oncoprotein is critically involved in both HTLV-1-replication and T-cell proliferation, a prerequisite to the development of ATLL. In this study, we investigated the in vivo contribution of the Tax PDZ domain-binding motif (PBM) to the lymphoproliferative process. To that aim, we examined T-cell proliferation in humanized mice (hu-mice) carrying a human hemato-lymphoid system infected with either a wild type (WT) or a Tax PBM-deleted (ΔPBM) provirus. We observed that the frequency of CD4+ activated T-cells in the peripheral blood and in the spleen was significantly higher in WT than in ΔPBM hu-mice. Likewise, human T-cells collected from WT hu-mice and cultivated in vitro in presence of interleukin-2 were proliferating at a higher level than those from ΔPBM animals. We next examined the association of Tax with the Scribble PDZ protein, a prominent regulator of T-cell polarity, in human T-cells analyzed either after ex vivo isolation or after in vitro culture. We confirmed the interaction of Tax with Scribble only in T-cells from the WT hu-mice. This association correlated with the presence of both proteins in aggregates at the leading edge of the cells and with the formation of long actin filopods. Finally, data from a comparative genome-wide transcriptomic analysis suggested that the PBM-PDZ association is implicated in the expression of genes regulating proliferation, apoptosis and cytoskeletal organization. Collectively, our findings suggest that the Tax PBM is an auxiliary motif that contributes to the sustained growth of HTLV-1 infected T-cells in vivo and in vitro and is essential to T-cell immortalization. The viral Tax oncoprotein is a critical contributor to the development of adult T-cell leukemia/lymphoma, an aggressive malignant proliferation of T lymphocytes. Tax contains a PDZ domain-binding motif (PBM) that favors the interaction with several cellular PDZ proteins. Here, we compare the in vivo involvement of the Tax PBM in humanized mice infected with either a full-length provirus or a Tax PBM-deleted provirus. We observe that the establishment of the sustained lymphoproliferation in the peripheral blood of infected mice is dependent on the Tax PBM. Furthermore, binding of the Tax PBM to the PDZ Scribble protein correlated with perturbations of cytoskeletal organization and cell polarity. In addition, genome-wide transcriptomic analyses strongly suggest that the association of Tax PBM with cellular PDZ proteins results in the expression of several genes involved in proliferation, apoptosis and cytoskeletal organization. Collectively, these results indicate that the Tax PBM is an auxiliary motif that contributes to the growth of HTLV-1 infected T-cells. As a consequence, targeting the PBM/PDZ nodes using small peptides may have the potential to antagonize the Tax-induced lymphoproliferation, offering a novel strategy for the treatment of this disease.

Isolated and identified more than 30 years ago, Human T-cell Leukemia Virus type 1 (HTLV-1) is the etiological agent of adult T-cell leukemia/lymphoma (ATL), an aggressive lymphoproliferative disease of activated CD4+ T cells, and other inflammatory disorders such as HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). A variety of animal models have contributed to the fundamental knowledge of HTLV-1 transmission, pathogenesis and to the design of novel therapies to treat HTLV-1 associated diseases. Small animal models (rabbits, rats, mice) as well as large animal models (monkeys) have been utilized to significantly advance characterization of the viral proteins and of virus-infected cells in the early steps of infection, as well as in the development of leukemogenic and immunopathogenic processes. Over the past two decades, the creation of new immuno-compromised mouse strains that are robustly reconstituted with a functional human immune system (HIS) after being transplanted with human tissues or progenitor cells has revolutionized the in vivo investigation of viral infection and pathogenesis. Recent observations obtained in HTLV-1-infected humanized HIS mice that develop lymphomas provide the opportunity to study the evolution of the proviral clonality in human T cells present in different lymphoid organs. Current progress in the improvement of those humanized models will favor the testing of drugs and the development of targeted therapies against HTLV-1-associated diseases.

The first discovered human retrovirus, Human T-Lymphotropic Virus type 1 (HTLV-1), is responsible for an aggressive form of T cell leukemia/lymphoma. Mouse models recapitulating the leukemogenesis process have been helpful for understanding the mechanisms underlying the pathogenesis of this retroviral-induced disease. This review will focus on the recent advances in the generation of immunodeficient and human hemato-lymphoid system mice with a particular emphasis on the development of mouse models for HTLV-1-mediated pathogenesis, their present limitations and the challenges yet to be addressed.