中华皮肤科杂志
中華皮膚科雜誌
중화피부과잡지
Chinese Journal of Dermatology
2014年
8期
574-577
,共4页
樊建勇%王颖%杨慧兰%梁洁%李翠华
樊建勇%王穎%楊慧蘭%樑潔%李翠華
번건용%왕영%양혜란%량길%리취화
疱疹病毒2型,人%树突细胞%单纯疱疹病毒疫苗
皰疹病毒2型,人%樹突細胞%單純皰疹病毒疫苗
포진병독2형,인%수돌세포%단순포진병독역묘
Herpesvirus 2,human%Dendritic cells%Herpes simplex virus vaccines
目的 观察单纯疱疹病毒2 gD(HSV-2 gD)基因的树突细胞(DC)疫苗在动物实验中诱发特异性免疫应答情况.方法 用腺病毒介导的、HSV-2 gD基因修饰的DC(pAdeno-HSV-2 gD-DC)疫苗免疫BALB/c小鼠3次,末次免疫后第10天检测小鼠血清中gD IgG水平;同时,分离小鼠脾淋巴细胞,用HSV-2gD蛋白刺激,MTT法检测小鼠脾淋巴细胞增殖情况、乳酸脱氢酶释放法测定细胞毒性T淋巴细胞(CTL)活性及ELISA法检测脾淋巴细胞培养上清中干扰素γ(IFN-γ)、白介素4(IL-4)水平.实验分为4组,pAdeno-DC组、pAdeno-HSV-2 gD-DC组、DC组、空白对照组,每组10只.结果 pAdeno-HSV-2 gD-DC组小鼠血清内可检测到高滴度针对HSV-2gD蛋白的抗体(A450值为0.313±0.034),与pAdeno-DC组(0.034±0.009)、DC组(0.028±0.009)、空白对照组(0.026±0.010)比较,差异有统计学意义(P＜0.05).pAdeno-HSV-2 gD-DC可以诱导小鼠脾淋巴细胞增殖(刺激指数为1.600土0.215)、有效杀伤靶细胞(CTL活性为37.1％),与pAdeno-DC组(分别为1.063±0.070和16.0％)、DC组(1.056±0.063和14.9％)、空白对照组(1.020±0.051和15.7％)比较,差异均有统计学意义(P＜0.05).pAdeno-HSV-2 gD-DC组小鼠分离的脾细胞培养上清可以检测到高水平IFN-γ(A450值为0.568±0.031)和IL-4(A450值为0.544±0.043),与pAdeno-DC组(0.266±0.021和0.278±0.037)、DC组(0.271±0.023和0.275±0.044)、空白对照组(0.252±0.012和0.245±0.051)比较,差异均有统计学意义(P＜0.05).结论 构建的pAdeno-HSV-2 gD-DC疫苗可以在小鼠中产生较强的特异性免疫应答.
目的 觀察單純皰疹病毒2 gD(HSV-2 gD)基因的樹突細胞(DC)疫苗在動物實驗中誘髮特異性免疫應答情況.方法 用腺病毒介導的、HSV-2 gD基因脩飾的DC(pAdeno-HSV-2 gD-DC)疫苗免疫BALB/c小鼠3次,末次免疫後第10天檢測小鼠血清中gD IgG水平;同時,分離小鼠脾淋巴細胞,用HSV-2gD蛋白刺激,MTT法檢測小鼠脾淋巴細胞增殖情況、乳痠脫氫酶釋放法測定細胞毒性T淋巴細胞(CTL)活性及ELISA法檢測脾淋巴細胞培養上清中榦擾素γ(IFN-γ)、白介素4(IL-4)水平.實驗分為4組,pAdeno-DC組、pAdeno-HSV-2 gD-DC組、DC組、空白對照組,每組10隻.結果 pAdeno-HSV-2 gD-DC組小鼠血清內可檢測到高滴度針對HSV-2gD蛋白的抗體(A450值為0.313±0.034),與pAdeno-DC組(0.034±0.009)、DC組(0.028±0.009)、空白對照組(0.026±0.010)比較,差異有統計學意義(P＜0.05).pAdeno-HSV-2 gD-DC可以誘導小鼠脾淋巴細胞增殖(刺激指數為1.600土0.215)、有效殺傷靶細胞(CTL活性為37.1％),與pAdeno-DC組(分彆為1.063±0.070和16.0％)、DC組(1.056±0.063和14.9％)、空白對照組(1.020±0.051和15.7％)比較,差異均有統計學意義(P＜0.05).pAdeno-HSV-2 gD-DC組小鼠分離的脾細胞培養上清可以檢測到高水平IFN-γ(A450值為0.568±0.031)和IL-4(A450值為0.544±0.043),與pAdeno-DC組(0.266±0.021和0.278±0.037)、DC組(0.271±0.023和0.275±0.044)、空白對照組(0.252±0.012和0.245±0.051)比較,差異均有統計學意義(P＜0.05).結論 構建的pAdeno-HSV-2 gD-DC疫苗可以在小鼠中產生較彊的特異性免疫應答.
목적 관찰단순포진병독2 gD(HSV-2 gD)기인적수돌세포(DC)역묘재동물실험중유발특이성면역응답정황.방법 용선병독개도적、HSV-2 gD기인수식적DC(pAdeno-HSV-2 gD-DC)역묘면역BALB/c소서3차,말차면역후제10천검측소서혈청중gD IgG수평;동시,분리소서비림파세포,용HSV-2gD단백자격,MTT법검측소서비림파세포증식정황、유산탈경매석방법측정세포독성T림파세포(CTL)활성급ELISA법검측비림파세포배양상청중간우소γ(IFN-γ)、백개소4(IL-4)수평.실험분위4조,pAdeno-DC조、pAdeno-HSV-2 gD-DC조、DC조、공백대조조,매조10지.결과 pAdeno-HSV-2 gD-DC조소서혈청내가검측도고적도침대HSV-2gD단백적항체(A450치위0.313±0.034),여pAdeno-DC조(0.034±0.009)、DC조(0.028±0.009)、공백대조조(0.026±0.010)비교,차이유통계학의의(P＜0.05).pAdeno-HSV-2 gD-DC가이유도소서비림파세포증식(자격지수위1.600토0.215)、유효살상파세포(CTL활성위37.1％),여pAdeno-DC조(분별위1.063±0.070화16.0％)、DC조(1.056±0.063화14.9％)、공백대조조(1.020±0.051화15.7％)비교,차이균유통계학의의(P＜0.05).pAdeno-HSV-2 gD-DC조소서분리적비세포배양상청가이검측도고수평IFN-γ(A450치위0.568±0.031)화IL-4(A450치위0.544±0.043),여pAdeno-DC조(0.266±0.021화0.278±0.037)、DC조(0.271±0.023화0.275±0.044)、공백대조조(0.252±0.012화0.245±0.051)비교,차이균유통계학의의(P＜0.05).결론 구건적pAdeno-HSV-2 gD-DC역묘가이재소서중산생교강적특이성면역응답.
Objective To evaluate the specific immune response induced by a dendritic cell-based adenovirus-mediated vaccine carrying the herpes simplex virus type 2 glycoprotein D gene (pAdeno-HSV-2 gD-DC) in BALB/c mice.Methods Forty BALB/c mice were equally divided into four groups:blank control group receiving no treatment,pAdeno-DC group immunized with pAdeno-DC,pAdeno-HSV-2 gD-DC group immunized with the previously constructed vaccine pAdeno-HSV-2 gD-DC,DC group immunized with DCs only.Totally,three rounds of vaccination were conducted at a 7-day interval.Ten days after the last vaccination,serum samples were collected and spleen cells were isolated from these mice.Enzyme-linked immunosorbent assay (ELISA) was performed to measure the level of IgG antibody against HSV-2 gD in the serum samples.Some spleen cells were stimulated with HSV-2 gD protein (10 mg/L) for 72 hours; then,ELISA was carried out to determine the levels of interferon (IFN)-γand interleukin (IL)-4 in the supernatant,and 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) assay to estimate the proliferative activity of these cells.The cytotoxicity of spleen cells was also evaluated based on the measurement of lactate dehydrogenase (LDH) release.Results The serum level of IgG antibody against HSV-2 gD (given in the absorbance value at 450 nm) was 0.313 ± 0.034 in the pAdeno-HSV-2 gD-DC group,significantly higher than that in the pAdeno-DC group,DC group and blank control group (0.034 ± 0.009,0.028 ± 0.009 and 0.026 ± 0.010 respectively,all P ＜ 0.05).Increased proliferative activity and cytotoxicity were observed in spleen cells from the pAdeno-HSV-2 gD-DC group compared with those from the pAdeno-DC group,DC group and blank control group (cell stimulation index:1.600 ± 0.215 vs.1.063 ± 0.070,1.056 ± 0.063 and 1.020 ± 0.051,all P ＜ 0.05; percentage of cytotoxicity:37.1％ vs.16.0％,14.9％ and 15.7％,all P ＜ 0.05).The levels of IFN-γ and IL-4 (both given in the absorbance value at 450 nm) were 0.568 ± 0.031 and 0.544-± 0.043 respectively in the supernatant of spleen cells from the pAdeno-HSV-2 gD-DC group,compared to 0.266 ± 0.021 and 0.278 ± 0.037 respectively in the pAdeno-DC group (bothP＜ 0.05),0.271 ± 0.023 and 0.275 ± 0.044 respectively in the DC group (bothP＜ 0.05),and 0.252 ± 0.012 and 0.245 ± 0.051 respectively in the blank control group (both P＜ 0.05).Conclusion The vaccine pAdenoHSV-2 gD-DC could induce a specific and strong immune response in BALB/c mice.
