中国病原生物学杂志
中國病原生物學雜誌
중국병원생물학잡지
JOURNAL OF PATHOGEN BIOLOGY
2006年
3期
161-163,封底
,共4页
刘向前%吴少庭%秦莉%袁仕善%黄达娜%雷明军%潘晖榕%林绮萍%张仁利%高世同%刘能保
劉嚮前%吳少庭%秦莉%袁仕善%黃達娜%雷明軍%潘暉榕%林綺萍%張仁利%高世同%劉能保
류향전%오소정%진리%원사선%황체나%뢰명군%반휘용%림기평%장인리%고세동%류능보
SARS%DNA疫苗%组织病理学
SARS%DNA疫苗%組織病理學
SARS%DNA역묘%조직병이학
Severe Acute Respiratory Syndrome(SARS)%DNA vaccine%histopathology
目的用组织病理学方法观察几种SARS DNA疫苗对BALB/c小鼠重要器官的影响,为研制安全有效的SARS DNA疫苗奠定基础. 方法用RT-PCR的方法从SARS冠状病毒(SARS-CoV)基因组中扩增出M片段、E片段、N片段和S基因的两个主要片段S1, S2,然后将这些基因片段分别亚克隆至pVAC真核表达载体,制备出几种SARS DNA疫苗pVAC-S1、 pVAC-S2、 pVAC-M、 pVAC-E、pVAC-N.用这些疫苗分别或联合免疫BALB/c小鼠后,取小鼠重要器官心、肝、脾、肺、肾,观察其组织病理学改变. 结果鼠心、脾和肾未见明显的组织病理学异常,但部分小鼠的肝和肺表现出下列病理变化:肝:出现片状肝细胞染色加深和肝细胞核固缩等凋亡早期改变,部分还可见到肝细胞水变性和肝窦变窄甚至消失等病变.肺:肺泡隔增厚,肺泡轮廓消失,或肺组织淤血水肿,甚至出现支气管肺炎改变.同时,在pVAC空质粒对照组也可见个别小鼠出现上述肝、肺病变. 结论由于对肝、肺产生组织病理学异常改变,制备高效、安全的SARS DNA疫苗还有待进一步研究和完善,载体的选择和质粒的用量也是必须加以考虑的问题.
目的用組織病理學方法觀察幾種SARS DNA疫苗對BALB/c小鼠重要器官的影響,為研製安全有效的SARS DNA疫苗奠定基礎. 方法用RT-PCR的方法從SARS冠狀病毒(SARS-CoV)基因組中擴增齣M片段、E片段、N片段和S基因的兩箇主要片段S1, S2,然後將這些基因片段分彆亞剋隆至pVAC真覈錶達載體,製備齣幾種SARS DNA疫苗pVAC-S1、 pVAC-S2、 pVAC-M、 pVAC-E、pVAC-N.用這些疫苗分彆或聯閤免疫BALB/c小鼠後,取小鼠重要器官心、肝、脾、肺、腎,觀察其組織病理學改變. 結果鼠心、脾和腎未見明顯的組織病理學異常,但部分小鼠的肝和肺錶現齣下列病理變化:肝:齣現片狀肝細胞染色加深和肝細胞覈固縮等凋亡早期改變,部分還可見到肝細胞水變性和肝竇變窄甚至消失等病變.肺:肺泡隔增厚,肺泡輪廓消失,或肺組織淤血水腫,甚至齣現支氣管肺炎改變.同時,在pVAC空質粒對照組也可見箇彆小鼠齣現上述肝、肺病變. 結論由于對肝、肺產生組織病理學異常改變,製備高效、安全的SARS DNA疫苗還有待進一步研究和完善,載體的選擇和質粒的用量也是必鬚加以攷慮的問題.
목적용조직병이학방법관찰궤충SARS DNA역묘대BALB/c소서중요기관적영향,위연제안전유효적SARS DNA역묘전정기출. 방법용RT-PCR적방법종SARS관상병독(SARS-CoV)기인조중확증출M편단、E편단、N편단화S기인적량개주요편단S1, S2,연후장저사기인편단분별아극륭지pVAC진핵표체재체,제비출궤충SARS DNA역묘pVAC-S1、 pVAC-S2、 pVAC-M、 pVAC-E、pVAC-N.용저사역묘분별혹연합면역BALB/c소서후,취소서중요기관심、간、비、폐、신,관찰기조직병이학개변. 결과서심、비화신미견명현적조직병이학이상,단부분소서적간화폐표현출하렬병리변화:간:출현편상간세포염색가심화간세포핵고축등조망조기개변,부분환가견도간세포수변성화간두변착심지소실등병변.폐:폐포격증후,폐포륜곽소실,혹폐조직어혈수종,심지출현지기관폐염개변.동시,재pVAC공질립대조조야가견개별소서출현상술간、폐병변. 결론유우대간、폐산생조직병이학이상개변,제비고효、안전적SARS DNA역묘환유대진일보연구화완선,재체적선택화질립적용량야시필수가이고필적문제.
Objective To observe the effect of several SARS DNA vaccines on vital organs of BALB/c mice by a histopathological method, so as to lay foundation for developing an effective and safe DNA vaccine against SARS. Methods The genes encoding M, E, N and S1, S2 (two main DNA fragments of S gene) were amplified from SARS-CoV genome RNA by RT-PCR method. Then several DNA vaccines, pVAC-S1, pVAC-S2, pVAC-M, pVAC-E, pVAC-N, were obtained by subcloning these DNA fragments into pVAC eukaryotic expression vector, respectively. After the BALB/c mice were immunized with these vaccines alone or in combination, the organs including heart, liver, spleen, lung and kidney were taken from the mice for histopathology study. Results No obvious changes were observed in heart, spleen and kidney, but part of the mice presented histopathological damage in liver and lung. In liver, the changes at an early stage of apoptosis or necrosis process, such as the cytoplasm deeper stained and the nuclei becoming denser in hepatocytes were found in some immunized mice. Hydropic degeneration and hepatic sinusoids becoming narrow or even disappeared were also observed in part of the hepatic tissue sections. In lung, thickening of alveolar septum and disintegration of alveolar structure, or lung edema and congestion, and even the pathological change of bronchial pneumonia were observed. However, pathological changes in liver and lung mentioned above could also be observed in a few mice treated with pVAC empty plasmid as a vector control. Conclusion On account of the toxicity in liver and lung, further investigation and advancement as well as appropriate vector and dosage should be concerned to develop effective and safe DNA vaccines against SARS.
