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Blood, Vol. 105, Issue 11, 4377-4382, June 1, 2005
Negative thymocyte selection to HERV-K18 superantigens in humans
Blood Meylan et al.
105: 4377
Supplemental materials for: Meylan et al, Vol 105, Issue 11, 4377-4382
Files in this Data Supplement:
- Figure S1. HERV-K18 SAg induced decrease in thymocyte cell numbers. (PDF, 35 KB) -
HERV-K18 SAg-induced negative selection resulted in a significant, and dose-dependent decrease in cell numbers, as compared to thymocytes cultured alone, or the vector control. The results from 5 independent experiments are shown, and significance levels are indicated (t-test, P < .05*; P < .01**).
- Figure S2. Immature and semimature CD4SP, but not mature SP CD4 and CD8 thymocytes, are deletion-sensitive. (PDF, 76 KB) -
(A) Purified immature DP, and purified mature SP thymocytes (not shown) were analyzed for their capacity to undergo negative selection upon culture with increasing doses of vector- and SAg-transfectants. The percentages of surviving DP thymocytes after culture are indicated. (B) The loss of function mutant
 53 is expressed as the wt SAg (upper panel). Two independent 53 transfectants, and a HERV-K18.1 wt SAg transfectant were analyzed by RPA with probes specific for the SAg, and for mouse TBP as control (lower panel). Expression of the transiently transfected wt and 53 HERV-K18.1 3×HA fusion proteins were detected by Western blot in HeLa cells. The HERV-K18.1 3×HA fusion protein retains SAg activity comparably to the HERV-K18.1 SAg (not shown).
- Figure S3. Differential SAg reactivity is a consistent phenotype. (PDF, 73 KB) -
(A) T cells from two donors that expanded SAg reactive T cells, but that had different starting levels of V
 7+ T cells were cultured with HERV-K18 SAg and control transfectants, and V expansions were analyzed by FACS. For these experiments the human B cell line TK6 was used. (B) T cells from individuals that were either weakly or strongly expanding were analyzed on two separate occasions spaced by months from each other, for their capacity to expand SAg reactive V7+ T cells (upper panel). Reactivity i), to the bacterial SAg SEB stimulating V12+ T cells (lower panel), and ii), to the polyclonal recall antigen tetanus toxoid (TT) was identical in both individuals (upper and lower panels).
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