Figure 5
Tumors characterization. (A) Phenotypic analysis of extranodal tumors developed in NOD/SCID mice injected with splenocytes from HIV Tg mouse #28. Flow analysis was performed in tumors from mice from groups injected with CD19+, B220+, CD93+, and the control CD3-. Mice shown are representative from each group. Tumors were analyzed for the following cell markers consisting in: CD3, CD19, CD43, CD93, B220, IgM, CD21, CD5, pre-BCR, Sca1, CD127, CD138, and CD34. “g1”and “g2” indicate the not activated and activated gates, respectively. “+”, “++”, “+++” indicate peak fluorescence intensity one, two or three logs higher than isotype control, respectively; “-” indicates no change in peak fluorescence intensity compared to isotype control; “+/−” indicates peak fluorescence intensity less than one log higher than isotype control. Flow histograms of extranodal tumors are shown in Additional 1: Figure S7. (B) Analysis of D-J rearrangement of the IgH gene. Genomic DNAs from extranodal tumors developed in NOD/SCID injected with B220+, CD19+, CD93+ and CD3- splenocytes purified from HIV Tg mouse n.28 were analyzed for D-J rearrangement of the IgH gene. Splenocytes from a wild type (WT) FVBN mouse were used as a control polyclonal B cells show four bands corresponding to D-JH1, D-JH2, D-JH3 and D-JH4, indicating that these four segments were rearranged, while tumors were monoclonal. The repertoire of Ig genes utilized in the monoclonal tumors indicated a preferential usage of the D-JH3 family of Ig H variable genes. (C) Molecular analysis of oncogenes expressed in extranodal tumors developed in NOD/SCID injected with B220+, CD19+, CD93+ and CD3- splenocytes purified from HIV Tg mouse n.28. A single mouse representative for each group of mice injected is shown. Oncogenes analysis was performed by SYBR Green semiquantitative real-time RT-PCR as described in material and methods.