重组白介素-4促进外周血淋巴细胞增殖和抗辐射的作用

Aim To demonstrate the promoting-proliferation effects of re-combinant human interleukin-4(rhIL-4)on lymphocyte subsets from normal indivaduals and its radiopro tection on lymphocytes irradiated b yγ-rays in vitro.Methods MTT colorimetric assay,immunohistochemical staining with alkaline phosphatase and TdT-mediated dUTP nick end la-beling(TUNEL)technique were used to detect the lym phocyte prolifera-tion,T and B lymphocyte subsets and t he lymphocyte apoptosis.Results(1)In the presence of adequate concentr ation of pokeweed mitogen(PWM),a definite concentration of rhIL-4could promote markedly the peripheral lymphocyte proliferati on of normal indivaduals,and displa yed stronger effects in serum-free cult ure system and 24h after culturing;(2)It is confirmed that the proliferati on of T cell subsets and B cells in-duced by rhIL-4might be major account for the proliferation of periphera llymphocytes.(3)A definite concentration of rhIL-4c ould obviously suppress the reduction of total lymp hocytes,T H ,T S subsets and B cells caused by 6Gy γ-irradiation.;and(4)It was also found that rhIL-4could inhibit distinctively abundant lymphocyte apoptosis induced by a -cute irradiation,suggesting that t his might be the major cause to alle-viate the radiation injury of lymphocytes and improve the immunological function of body.Conclusion rhIL-4can increase the number of pe-ripheral T and B lymphocytes from normal indivaduals,and demon-strated evident inhibition on the ly mphocytic reduction induced by leth al dose irradiation.

大鼠骨髓来源树突状细胞体外培养体系的优化

目的探讨优化大鼠骨髓来源树突状细胞(DC)体外培养体系的方法。方法应用重组大鼠粒细胞-巨噬细胞集落刺激因子(recombinant rat GM-CSF,rrGM-CSF)20μg/L和重组大鼠白介素(recombinant rat interleukin,rrIL)-410μg/L诱导分化Lewis大鼠骨髓单个核细胞,获得未成熟DC(immature DC,imDC)。第6日加入终含量为100μg/L的脂多糖(lipopolysaccharides,LPS)刺激细胞成为成熟DC(mature DC,mDC)。用倒置相差显微镜观察细胞形态,流式细胞术鉴定细胞表型、双抗夹心酶联免疫吸附测定法(ELISA)检测DC培养上清液中白介素(IL)-12、IL-10的含量,并以Brown Norway大鼠脾脏T淋巴细胞为反应细胞,做混合淋巴细胞反应检测,观察DC刺激T淋巴细胞增殖能力。结果经细胞形态学、流式细胞术及混合淋巴细胞反应三方面鉴定,证实所培养细胞为DC。体外培养第6日,见大量增殖细胞集落,细胞高表达OX62。LPS刺激细胞后,细胞伪足样突起明显增多,细胞逐渐呈半悬浮、悬浮生长,细胞表面高表达主要组织相容性抗原复合体(MHC)Ⅱ类分子和共刺激分子(CD40、CD86)。与imDC比较,mDC分泌IL-10和IL-12显著增加,刺激同种异体T淋巴细胞增殖的能力显著提高。结论通过采用调整rrGM-CSF和rrIL-4的用量等方法优化体外培养体系诱导大鼠骨髓单个核细胞,可分化出大量骨髓源性DC。

Immunogenicity analysis following human immunodeficiency virus recombinant DNA and recombinant vaccinia virus Tian Tan prime-boost immunization

This study assessed and compared the immunogenicity of various immunization strategies in mice using combinations of re- combinant DNA （pCCMp24） and recombinant attenuated vaccinia virus Tian Tan （rddVTT_ccMpe4）. Intramuscular immuniza- tion was performed on days 0 （prime） and 21 （boost）. The immunogenicity of the vaccine schedules was determined by meas- uring human immunodeficiency virus （HIV）-specific binding antibody levels and cytokine （interleukin-2 and interleukin-4） concentrations in peripheral blood, analyzing lymphocyte proliferation capacity against HIV epitopes and CD4~/CD8＋cell ratio, and monitoring interferon-gamma levels at different times post-immunization. The results showed that pCCMp24, rddVTT.ccMp24 and their prime-boost immunization induced humoral and cellular immune responses. The pCCMp24/ rddVTT.ccMp24 immunization strategy increased CD8＋ T cells and induced more IFN-7-secreting cells compared with sin- gle-shot rDNA. The prime-boost immunization strategy also induced the generation of cellular immunological memory to HIV epitope peptides. These results demonstrated that prime-boost immunization with rDNA and rddVTT_ccMp24 had a tendency to induce greater cellular immune response than single-shot vaccinations, especially IFN-7 response, providing a basis for further studies.