Figure 1
Figure 1. ATMKO.CD3εKO mice develop early-onset B-cell lymphomas in spleen and liver that resemble DLBCL. (A) Representative image of spleens (right) and livers (left) from ATMWT.CD3εKO and ATMKO.CD3εKO mice. (B) Hematoxylin and eosin staining showing spleen and liver sections from ATMWT.CD3εKO (top) and ATMKO.CD3εKO mice (bottom) (original magnification ×10; bar represents 200 μm). (C) Hematoxylin and eosin staining of a spleen section prepared from a tumor-bearing ATMKO.CD3εKO mouse (original magnification ×1000; bar represents 20 μm). Red and black arrows denote centroblasts and immunoblasts, respectively. (D) B220 staining of a spleen section from a tumor-bearing ATMKO.CD3εKO mouse (original magnification ×400; bar = 100 μm) using 3,3′-diaminobenzidine tetrahydrochloride as chromogen. Images were viewed with an Olympus BX41 microscope and photographed with an Olympus DP71 camera. Dynamic positioning controller software (version 3.3.1.292) was used for image acquisition. (E) The probability of dying with a tumor was determined using the Kaplan-Meier method. Kaplan-Meier analysis of tumor incidence studies in ATMKO.CD3εKO (n = 25) and ATMKO (n = 24) mice. ATMKO.CD3εKO mice died with B-cell lymphomas and ATMKO mice died with thymic T-cell lymphomas. No thymic T-cell lymphomas were observed in ATMKO.CD3εKO mice and no B-cell lymphomas were detected in ATMKO mice. No ATMWT.CD3εKO or ATMWT mice died with lymphomas. Mice euthanized and not found to have either a B- or T-cell tumor had their observations censored in the analysis. (F) Flow cytometric analysis of IgM, B220, CD40, MHC class I (MHCI; Kb), MHC class II (MHCII; IAb), CD95, and CD86 expression (open histogram) or isotype control staining (shaded histogram) on normal splenic B cells (gated on IgM+ cells) and on 1 representative ATMKO.CD3εKO B-cell lymphoma. (G) CD21 and CD23 expression on splenic B220+ B cells and 1 ATMKO.CD3εKO B-cell lymphoma. These histograms are representative of results obtained from 20 individual tumors. Data acquisition was performed using a FACSCaliber flow cytometer (BD Biosciences), and FlowJo software (Tree Star) was used for analysis.

ATMKO.CD3εKO mice develop early-onset B-cell lymphomas in spleen and liver that resemble DLBCL. (A) Representative image of spleens (right) and livers (left) from ATMWT.CD3εKO and ATMKO.CD3εKO mice. (B) Hematoxylin and eosin staining showing spleen and liver sections from ATMWT.CD3εKO (top) and ATMKO.CD3εKO mice (bottom) (original magnification ×10; bar represents 200 μm). (C) Hematoxylin and eosin staining of a spleen section prepared from a tumor-bearing ATMKO.CD3εKO mouse (original magnification ×1000; bar represents 20 μm). Red and black arrows denote centroblasts and immunoblasts, respectively. (D) B220 staining of a spleen section from a tumor-bearing ATMKO.CD3εKO mouse (original magnification ×400; bar = 100 μm) using 3,3′-diaminobenzidine tetrahydrochloride as chromogen. Images were viewed with an Olympus BX41 microscope and photographed with an Olympus DP71 camera. Dynamic positioning controller software (version 3.3.1.292) was used for image acquisition. (E) The probability of dying with a tumor was determined using the Kaplan-Meier method. Kaplan-Meier analysis of tumor incidence studies in ATMKO.CD3εKO (n = 25) and ATMKO (n = 24) mice. ATMKO.CD3εKO mice died with B-cell lymphomas and ATMKO mice died with thymic T-cell lymphomas. No thymic T-cell lymphomas were observed in ATMKO.CD3εKO mice and no B-cell lymphomas were detected in ATMKO mice. No ATMWT.CD3εKO or ATMWT mice died with lymphomas. Mice euthanized and not found to have either a B- or T-cell tumor had their observations censored in the analysis. (F) Flow cytometric analysis of IgM, B220, CD40, MHC class I (MHCI; Kb), MHC class II (MHCII; IAb), CD95, and CD86 expression (open histogram) or isotype control staining (shaded histogram) on normal splenic B cells (gated on IgM+ cells) and on 1 representative ATMKO.CD3εKO B-cell lymphoma. (G) CD21 and CD23 expression on splenic B220+ B cells and 1 ATMKO.CD3εKO B-cell lymphoma. These histograms are representative of results obtained from 20 individual tumors. Data acquisition was performed using a FACSCaliber flow cytometer (BD Biosciences), and FlowJo software (Tree Star) was used for analysis.

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