AIM:To purify the heat shock protein(HSP)70-associatedtumor peptides and to observe its non-MHC-I moleculerestrictive antitumor effect.METHODS:By ConA-sepharose affinity chromatography,ADP-agarose affinity chromatography,and DEAE anionexchange chromatography,we were able to purify HSP70-associated peptides from mouse hepatoma(HCaF)cellstreated in heat shock at 42℃.Specific active immunizationand adoptive cellular immunization assay were adopted toobserve the immunoprotective effect elicited by HSP70-associated peptide complexes isolated from HcaF.RESULTS:The finally purified HSP-associated peptides hada very high purity and specificity found by SDS-PAGE andWestern blot.Mice immunized with HSP70-associated peptidecomplexes purified from HCaF cells were protected fromHCaF living cell challenge.This effect was dose dependent.Adoptive immunization of immune spleen cells of miceimmunized with HSP70-associated peptide complexes couldelicit immunity against HCaF challenge,and the tumor-freemice could resist repeated challenges.This effect could becontinuously enhanced by repeated challenge with HCaFliving cells.The tumor-free mice could tolerate the challengefor as high as 1×10~7 HCaF cells.The mice immunized oncewith spleen cells pulsed with HSP70-associated peptidecomplexes in vitro could also result in a certain adoptiveimmunity against HCaF.CONCLUSION:High purity and specificity of HSP70-associated peptides could be achieved from tumor cellsby the low-pressure affinity chromatography methodused in this study.HSP70-associated peptide complexesderived from the HCaF can elicit non-MHC-I moleculerestrictive immunoprotective effect against HCaF.Thiseffect can be transferred by adoptive immunization tomice and enhanced by repeated challenge with HCaFlive cells. AIM: To purify the heat shock protein (HSP) 70-associated tumor peptides and observe its non-MHC-I molecule-specific antitumor effect. METHODS: By ConA-sepharose affinity chromatography, and ADAE-anion exchange chromatography, and were able to purify HSP70-associated peptides from mouse hepatoma (HCaF) cellstreated in heat shock at 42 ° C. Specific active immunization and adoptive cellular immunization assay were toobserve the immunoprotective effect elicited by HSP70-associated peptide complexes isolated from HcaF. RESULTS: The finally purified HSP-associated peptides had a very high purity and specificity found by SDS-PAGE and Western blot. Mice immunized with HSP70-associated peptide complexes purified from HCaF cells were protected from HCaF living cell challenge. This effect was dose dependent. Adaptive immunization of immune spleen cells of mice immunized with HSP70-associated peptide complexes couldelicit immunity against HCaF challenge, and the tumor-free mice could resist repeated challenges.This effect could becontinuously enhanced by repeated challenge with HCaFliving cells.The tumor-free mice could tolerate the challenge as as high as 1 × 10 ~ 7 HCaF cells.The mice immunized once with spleen cells pulsed with HSP70-associated peptide complexes in vitro could could result in a certain adoptive immunity against HCaF. CONCLUSION: High purity and specificity of HSP70-associated peptides could be achieved from tumor cells by the low-pressure affinity chromatography methodused in this study. HSP70-associated peptide complexes derived from the HCaF can elicit non-MHC-I moleculerestrictive immunoprotective effect against HCaF.Thiseffect can be transferred by adoptive immunization tomice and enhanced by repeated challenge with HCaFlive cells.