Objective To describe the phenotype of NK cells in Bama miniature pigs, and establish an efficient activation and culture method for porcine cytokine-induced killer ( CIK) cells in vitro. Methods The porcine peripheral blood mononuclear cells ( PBMCs) were isolated by Percoll gradient centrifugation, and the phenotype of NK cells was test-ed by detecting the CD2 + /CD8 + /CD3 - cell compartment. To establish an efficient activation and culture method for por-cine CIK cells, we optimized the culture conditions to improve the CIK activation efficiency. Results Using the optimized induction culture conditions, the ratio of CIK ( CD2 + /CD8 + /CD3 -) cells was up to 43. 63% at the fifth day, approxi-mately 5. 59 times increased compared with the initially separated PBMCs. Cell proliferation experiments showed that three obvious fluorescence peaks were observed on the fifth day. The results indicated that the induced CIK cells underwent three times cell division, in theory, about increased 8-fold compared with the initial separation of PBMCs. Furthermore, the qRT-PCR result of the surface markers of porcine NK cells also showed a similar variation tendency as the flow cytometry results. Conclusions Our findings demonstrate the successful establishment of an efficient activation and culture method for porcine CIK cells in vitro.

Objective To transfect the recombinant mIL-28A adenovirus vector into lung adenocarcinoma cell line LA795 and research its anticancer activity. Methods Transfected the constructed mouse IL-28(mIL-28) recombinant adenovirus vector into LA795 cell line, detected with PCR, immunocytal fluorescence, Tunel, Annexin V and MTT. Results Transfected with rAd-mlL-28A into the LA795 cells, mIL-28A gene expression products mRNA increased obviously, IL-28 expression was detected in cells obviously,apoptosis cell number increased, and the growth of LA795 cells transfected with rAd-mIL-28A were inhibited obviously. Conclusion The recombinant miL-28A adenovirus vector we have constructed, which expresses IL-28 when transfected to lung adenocarcinoma cell line LA795, inhibits growth of carcinoma cell to some extent, and may work by promoting the apoptosis of cancer cells.