by Prabal Banerjee, Adam Tripp, Michael D

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Adult T-cell leukemia/lymphoma development in HTLV-1–infected humanized SCID mice by Prabal Banerjee, Adam Tripp, Michael D. Lairmore, Lindsey Crawford, Michelle Sieburg, Juan Carlos Ramos, William Harrington, Mark A. Beilke, and Gerold Feuer Blood Volume 115(13):2640-2648 April 1, 2010 ©2010 by American Society of Hematology

Lymphomagenesis in HTLV-1 HU-NOD/SCID mice. Lymphomagenesis in HTLV-1 HU-NOD/SCID mice. Representative histologic analysis of HTLV-1-HU-NOD/SCID mice and mock mice killed 14 to 16 weeks after reconstitution. Development of lymphomas localized to the mesenteric (MLN) and lymph nodes surrounding the pancreas (PLN; red arrows) and spleen (A-B,D) in HTLV-1-HU-NOD/SCID mice in comparison with mock-infected HU-NOD/SCID mice (C,E). H&E staining of MLN (F), pancreatic lymph nodes (G), and infiltrating lymphocytes in spleen (I) of HTLV-1-HU-NOD/SCID mice in contrast to lymph nodes (H) and spleen (J) of mock-infected HU-NOD/SCID mice. H&E staining shows diffuse large scale lymphomas in the kidney, pancreas, liver, and lungs (K,M,O,Q) of HTLV-1-HU-NOD/SCID mice in comparison with kidney, pancreas, liver, and lungs (L,N,P,R) of mock-infected HU-NOD/SCID mice. Insets (K,M,O,Q) show immunohistochemical analysis for human CD45. Prabal Banerjee et al. Blood 2010;115:2640-2648 ©2010 by American Society of Hematology

Phenotype analysis of lymphomas from HTLV-1-HU-NOD/SCID and Tax1-HU-NOD/SCID mice. Phenotype analysis of lymphomas from HTLV-1-HU-NOD/SCID and Tax1-HU-NOD/SCID mice. Phenotype analysis of lymphocytes from the MLN (A,E,I,L) and the spleen (B,F,J,M) of HTLV-1-HU-NOD/SCID mouse (#7762, 16 weeks after injection) and MLN of Tax1-HU-NOD/SCID (#7749, 14 weeks after injection) mouse (C,G,K,N). Human CD3+ cells are gated and analyzed in panels I, J, and K. CD4+/CD8− are the predominant populations in HTLV-1-HU-NOD/SCID mice MLN (I) and spleen (J) and Tax1-HU-NOD/SCID mice MLN (K). Human lymphocytes in the mesenteric lymph nodes (D) of mock-infected donor-tissue matched HU-NOD/SCID (#7762, 16 weeks after injection) mice are predominantly of CD19+ (B-cell) lineage (H). HTLV-1 p19gag expression in CD4+ T cells from the MLN (L) and spleen (M) of the HTLV-1-HU-NOD/SCID mouse. Expression of GFP in the CD4+ T cells from (N) the MLN of the Tax1-HU-NOD/SCID mouse and (O) in cells from the MLN of mock-infected mouse. (P) Proliferation of CD4+/CD8− subpopulation among gated human lymphocytes in the thymus of the HTLV-1-HU-NOD/SCID mouse (#7241, 9 weeks after injection; iii) in comparison with mock HU-NOD/SCID mouse (#7241, 8 weeks after injection; ii). Comparative Giemsa staining of lymphoma cells derived from HTLV-1-HU-NOD/SCID mouse (Q) and RV-ATL tumor cell line (R).28 Prabal Banerjee et al. Blood 2010;115:2640-2648 ©2010 by American Society of Hematology

Hyperproliferation of infected HSCs in the BM of HTLV-1-HU-NOD/SCID mice. Hyperproliferation of infected HSCs in the BM of HTLV-1-HU-NOD/SCID mice. (A) Predominance of CD34+/CD38− human stem cell subpopulation among human lymphocytes in the BM of the HTLV-1-HU-NOD/SCID mouse (iii) in comparison with mock-infected HU-NOD/SCID mouse (ii) when gated on human CD45 subpopulation (i). (B) Cumulative quantization of CD34+/CD38− and CD34+/CD38+ subpopulations in BM of HTLV-1 infected (n = 7) and mock-infected HU-NOD/SCID mice (n = 8). Prabal Banerjee et al. Blood 2010;115:2640-2648 ©2010 by American Society of Hematology

HTLV-1 infection skews hematopoiesis to the T-cell lineage in HU-NSG mice. HTLV-1 infection skews hematopoiesis to the T-cell lineage in HU-NSG mice. Expression of CD19 and CD3 on lymphocytes (gated on human CD45) from the BM (A-B), thymus (C-D), spleen (E-F), and MLN (G-H) of HTLV-1-HU-NSG mouse (14 weeks after injection) in comparison with mock-infected HU-NSG mouse reconstituted with tissue-matched CD34+ donor cells. Human CD3+ cells were then gated and subsequently analyzed for CD4 and CD8 expression (i-viii). CD4+/CD8− single-positive T cells are the predominant population in HTLV-1-HU-NSG mice BM (ii), thymus (iv), spleen (vi), and MLN (viii) in comparison with mock-infected HU-NSG mice. Human lymphocytes in mock-infected donor tissue–matched HU-NSG mice (14 weeks after injection) mice show broader and more diverse hematopoietic lineage development, including the presence of B cells (CD19+), mature single CD8+ T cells, and immature CD4+/CD8+ T cells (i,iii,v,vii). Prabal Banerjee et al. Blood 2010;115:2640-2648 ©2010 by American Society of Hematology

Hyperproliferation of CD3+ T cells in HTLV-1/HU-NSG mice. Hyperproliferation of CD3+ T cells in HTLV-1/HU-NSG mice. Representative histologic analysis of the spleen of HTLV-1/HU-NSG mice and mock HU-NSG mice killed 14 weeks after reconstitution. (A-B) Immunohistochemical analysis of human CD3 expression in the spleen of HTLV-1-HU-NSG mice in comparison with mock-infected HU-NSG mice (×40 magnification). (C) Quantification of hyperproliferation of CD3+ T cells in the BM, thymus, spleen, and MLN of HTLV-1-HU-NSG mice in comparison with mock mice reconstituted with the same donor CD34+ cell preparation (HTLV-1, n = 6; mock, n = 4). Prabal Banerjee et al. Blood 2010;115:2640-2648 ©2010 by American Society of Hematology

Serial transplantation and engraftment of lymphomas cells from HTLV-1-HU-NOD/SCID mice. Serial transplantation and engraftment of lymphomas cells from HTLV-1-HU-NOD/SCID mice. (A) Pooled cells from the spleen and MLNs of HTLV-1-HU-NOD/SCID mouse (#7749) were serially transplanted intraperitoneally into naive NOD/SCID mice. Cells from lymphomas in the mesenteric lymph nodes (B), in the secondary recipient mice (#7749-2) as visualized by H&E (C), and immunohistochemistry staining for human CD45 (Hu-CD45; D). Phenotypic analysis of lymphoma cells in the MLN of secondary recipients demonstrate CD4+ staining (E), coexpression of HTLV-1 gag antigen p19gag (F), and expression of CD25 (G). H&E staining demonstrating diffuse large-scale lymphomas in the spleen (H), kidney (I), and small intestine (J) of secondary recipient NOD/SCID mice. (K) Immunohistochemical analysis for human CD45 in the spleen of secondary recipient mice that developed lymphoma. Prabal Banerjee et al. Blood 2010;115:2640-2648 ©2010 by American Society of Hematology