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Blood, Vol. 113, Issue 23, 5920-5926, June 4, 2009
Circulating clonotypic B cells in classic Hodgkin lymphoma
Blood Jones et al.
113: 5920
Supplemental materials for: Jones et al
Files in this Data Supplement:
- Figure S1. CD20+ B cells, but not HRS cells, generate and maintain HL cell lines (JPG, 61.3 KB)
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Varying numbers of the CD20+ and CD30+ cells from both L428 or KM-H2 B-cell cell lines were directly sorted into microtiter wells and then serially replated every seven days. As few as ten CD20+ cells from both cell lines expanded over seven days to over 103 cells containing both the B-cell and RS subpopulations. Serial replating as few as ten L428 and KM-H2 cells from the wells originally seeded with ten CD20+ cells generated cellular expansion for least four serial replatings. No wells seeded with one or ten sorted larger-sized CD30+ HRS cells from either cell line grew. Seeding 100 HRS cells from either cell line resulted in cellular expansion over seven days, but the expanded cells never contained the B-cell population. Moreover, no further cellular expansion was seen with the second replating of 100 HRS cells from either cell line. Three separate experiments yielded exactly the same findings.
- Figure S2. Lack of light chain restriction in the circulating CD27+ALDHhigh B cells from a newly-diagnosed HL patient (JPG, 54.9 KB)
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CD19+ cells were isolated from a newly-diagnosed HL patient and stained with Aldefluor and monoclonal antibodies against CD27 and (A) kappa and (B) lambda light chains.
- Figure S3. Circulating light chain-restricted CD27+ALDHhigh B cells show identical immunoglobulin rearrangements to HRS cells (JPG, 87.5 KB)
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(A) Clonal immunogloblulin kappa sequence traces from peripheral blood light chain-restricted B cells (top) and microdissected HRS cells (bottom) isolated from the patient shown in Fig. 6B,D. (B) The resulting sequences are nearly identical to each other (68/71 evaluable bases, with 2 of the differences in low quality regions) and to the IgK variable region (Giudicelli et al, Nucleic Acids Res. 2005;33:D256–D261. The Fr3 and CDR3 are indicated. Dots indicate identity with the germline IgKV1–8 sequence.
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