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. Author manuscript; available in PMC: 2010 Mar 15.

Published in final edited form as: Clin Cancer Res. 2009 Mar 10;15(6):1954–1963. doi: 10.1158/1078-0432.CCR-08-1290

(A and B) SPANX-B –specific CD4⁺ T cell lines were generated by repeated stimulations of human T cells with irradiated autologous iDC treated with SPANX-B protein. The immunodominant region of SPANX-B is located in the overlapping portion of synthetic peptides Pep-G1 and Pep-5, as the T cell line can be also activated to secrete IFNγ with irradiated mDCs pulsed with 1μg/ml Pep-G1 or Pep-5, but not with individual peptides (Pep-G2, Pep-6) or mixture of peptides (Pep-1, -2, -3, -4) specific to other regions of SPANX-B. The CD4⁺ T cell lines generated to SPANX-B protein (B), or Pep-5 (C), or Pep-G1 (D) specifically and reciprocally recognize DCs pulsed with titrated amounts (μg/ml) of SPANX-B protein, or Pep-5, or Pep-G1, or Pep-9 and secrete IFNγ (pg/ml). The T cells were not activated with DCs pulsed with control murine class II peptide (MOPC) or with scrambled Pep-9 (Pep-9-Mod, D). **P<0.01 and ***P<0.001 value is for comparison with the group indicated by line. Shown, mean ± SEM of representative and reproducible results of at least three independent experiments performed in triplicates.

(A and B) SPANX-B –specific CD4⁺ T cell lines were generated by repeated stimulations of human T cells with irradiated autologous iDC treated with SPANX-B protein. The immunodominant region of SPANX-B is located in the overlapping portion of synthetic peptides Pep-G1 and Pep-5, as the T cell line can be also activated to secrete IFNγ with irradiated mDCs pulsed with 1μg/ml Pep-G1 or Pep-5, but not with individual peptides (Pep-G2, Pep-6) or mixture of peptides (Pep-1, -2, -3, -4) specific to other regions of SPANX-B. The CD4⁺ T cell lines generated to SPANX-B protein (B), or Pep-5 (C), or Pep-G1 (D) specifically and reciprocally recognize DCs pulsed with titrated amounts (μg/ml) of SPANX-B protein, or Pep-5, or Pep-G1, or Pep-9 and secrete IFNγ (pg/ml). The T cells were not activated with DCs pulsed with control murine class II peptide (MOPC) or with scrambled Pep-9 (Pep-9-Mod, D). **P<0.01 and ***P<0.001 value is for comparison with the group indicated by line. Shown, mean ± SEM of representative and reproducible results of at least three independent experiments performed in triplicates.