中华检验医学杂志
中華檢驗醫學雜誌
중화검험의학잡지
CHINESE JOURNAL OF LABORATORY MEDICINE
2011年
2期
158-163
,共6页
邱趁丽%黄相刚%卫军%乔晓春%仇超%万延民%王万海%张晓燕%徐建青
邱趁麗%黃相剛%衛軍%喬曉春%仇超%萬延民%王萬海%張曉燕%徐建青
구진려%황상강%위군%교효춘%구초%만연민%왕만해%장효연%서건청
HIV感染%HIV-1%T淋巴细胞,细胞毒性%基因组%免疫酶技术
HIV感染%HIV-1%T淋巴細胞,細胞毒性%基因組%免疫酶技術
HIV감염%HIV-1%T림파세포,세포독성%기인조%면역매기술
HIV infections%HIV-1%T-lymphocytes,cytotoxic%Genome%Immunoenzyme techniques
目的 分析研究不同感染时间和不同途径感染HIV/AIDS感染者覆盖全基因组的CTL应答特征.方法 将75例HIV/AIDS感染者分为3组,血液感染1～2年组(10例)、血液感染＞10年组(43例)和性接触感染1～2年组(22例);以HIV-1 B亚型构建全基因组17个肽库作为抗原,采用ELISpot检测各组HIV特异性CTL应答;采用流式细胞术检测各组CD4计数;采用实时定量RT-PCR检测各组HIV病毒载量.结果 血液感染1～2年组、血液感染＞10年组和性接触感染1～2年组感染者对HIV-B亚型17个肽库的平均应答频率分别为40%、65%、23%,经单向方差分析,3组感染者对HIV-1 B亚型17个肽库的应答频率差异有统计学意义(F=19.96,P＜0.01);3组感染者对HIV-B亚型17个肽库总应答强度范围分别为0～5 835 SFCs/106 PBMC、0～7 225 SFCs/106PBMC、0～9 740 SFCs/106PBMC,且3组感染者对HIV-B亚型17个肽库的应答强度差异亦有统计学意义(H=101.90,P＜0.01);此外,3组感染者对HIV-1 B亚型17个肽库的应答广度分别为7(2～11)个、11(9～14)个、4(2～6)个,3组感染者应答广度的差异也有统计学意义(H=34.75,P＜0.01).3组感染者应答频率、应答强度和应答广度从高到低依次为血液感染＞10年组、血液感染1～2年组、性接触感染1～2年组.不同感染时间和不同感染途径的感染者对Nef肽库和Gag肽库的应答百分比和应答强度均高于其他肽库.CD4计数＜200/μl、CD4计数为200～500/μl和≥500/μl3组感染者对17个肽库的总应答强度范围分别为0～18 475 SFCs/106 PBMC、350～34 095 SFCs/106PBMC、490～21 550 SFCs/106 PBMC,但差异无统计学意义(H=2.93,P=0.23);3组感染者CTL应答广度分别为3(0～8)个、10(2～17)个、10(1～17)个,3组间差异有统计学意义(H=14.72,P＜0.01),CD4计数＜200/μl的感染者CTL应答广度低于CD4计数为200～500/μl和≥500/μl组.不同病毒载量3组(＜LDL、LDL-1×104拷贝/ml和≥1×104拷贝/ml)标本对17个肽库的总应答强度范围分别为490～18 475 SFCs/106PBMC、0～24 115 SFCs/106PBMC、770～34 095 SFCs/106 PBMC,但3组间差异无统计学意义(H=0.79,P=0.67);应答广度分别为8(1～17)个、11(0～17)个、8(1～16)个,3组间差异也无统计学意义(H=5.27,P=0.07).结论 中国HIV/AIDS感染者中CTL应答多集中在Nef和Gag,这两个抗原为HIV/AIDS感染的优势抗原;感染时间和感染途径对CTL应答可产生显著影响;随感染时间的延长,免疫反应的强度与应答比例均增加.这些信息对设计针对中国人群的AIDS疫苗有较重要意义.
目的 分析研究不同感染時間和不同途徑感染HIV/AIDS感染者覆蓋全基因組的CTL應答特徵.方法 將75例HIV/AIDS感染者分為3組,血液感染1～2年組(10例)、血液感染＞10年組(43例)和性接觸感染1～2年組(22例);以HIV-1 B亞型構建全基因組17箇肽庫作為抗原,採用ELISpot檢測各組HIV特異性CTL應答;採用流式細胞術檢測各組CD4計數;採用實時定量RT-PCR檢測各組HIV病毒載量.結果 血液感染1～2年組、血液感染＞10年組和性接觸感染1～2年組感染者對HIV-B亞型17箇肽庫的平均應答頻率分彆為40%、65%、23%,經單嚮方差分析,3組感染者對HIV-1 B亞型17箇肽庫的應答頻率差異有統計學意義(F=19.96,P＜0.01);3組感染者對HIV-B亞型17箇肽庫總應答彊度範圍分彆為0～5 835 SFCs/106 PBMC、0～7 225 SFCs/106PBMC、0～9 740 SFCs/106PBMC,且3組感染者對HIV-B亞型17箇肽庫的應答彊度差異亦有統計學意義(H=101.90,P＜0.01);此外,3組感染者對HIV-1 B亞型17箇肽庫的應答廣度分彆為7(2～11)箇、11(9～14)箇、4(2～6)箇,3組感染者應答廣度的差異也有統計學意義(H=34.75,P＜0.01).3組感染者應答頻率、應答彊度和應答廣度從高到低依次為血液感染＞10年組、血液感染1～2年組、性接觸感染1～2年組.不同感染時間和不同感染途徑的感染者對Nef肽庫和Gag肽庫的應答百分比和應答彊度均高于其他肽庫.CD4計數＜200/μl、CD4計數為200～500/μl和≥500/μl3組感染者對17箇肽庫的總應答彊度範圍分彆為0～18 475 SFCs/106 PBMC、350～34 095 SFCs/106PBMC、490～21 550 SFCs/106 PBMC,但差異無統計學意義(H=2.93,P=0.23);3組感染者CTL應答廣度分彆為3(0～8)箇、10(2～17)箇、10(1～17)箇,3組間差異有統計學意義(H=14.72,P＜0.01),CD4計數＜200/μl的感染者CTL應答廣度低于CD4計數為200～500/μl和≥500/μl組.不同病毒載量3組(＜LDL、LDL-1×104拷貝/ml和≥1×104拷貝/ml)標本對17箇肽庫的總應答彊度範圍分彆為490～18 475 SFCs/106PBMC、0～24 115 SFCs/106PBMC、770～34 095 SFCs/106 PBMC,但3組間差異無統計學意義(H=0.79,P=0.67);應答廣度分彆為8(1～17)箇、11(0～17)箇、8(1～16)箇,3組間差異也無統計學意義(H=5.27,P=0.07).結論 中國HIV/AIDS感染者中CTL應答多集中在Nef和Gag,這兩箇抗原為HIV/AIDS感染的優勢抗原;感染時間和感染途徑對CTL應答可產生顯著影響;隨感染時間的延長,免疫反應的彊度與應答比例均增加.這些信息對設計針對中國人群的AIDS疫苗有較重要意義.
목적 분석연구불동감염시간화불동도경감염HIV/AIDS감염자복개전기인조적CTL응답특정.방법 장75례HIV/AIDS감염자분위3조,혈액감염1～2년조(10례)、혈액감염＞10년조(43례)화성접촉감염1～2년조(22례);이HIV-1 B아형구건전기인조17개태고작위항원,채용ELISpot검측각조HIV특이성CTL응답;채용류식세포술검측각조CD4계수;채용실시정량RT-PCR검측각조HIV병독재량.결과 혈액감염1～2년조、혈액감염＞10년조화성접촉감염1～2년조감염자대HIV-B아형17개태고적평균응답빈솔분별위40%、65%、23%,경단향방차분석,3조감염자대HIV-1 B아형17개태고적응답빈솔차이유통계학의의(F=19.96,P＜0.01);3조감염자대HIV-B아형17개태고총응답강도범위분별위0～5 835 SFCs/106 PBMC、0～7 225 SFCs/106PBMC、0～9 740 SFCs/106PBMC,차3조감염자대HIV-B아형17개태고적응답강도차이역유통계학의의(H=101.90,P＜0.01);차외,3조감염자대HIV-1 B아형17개태고적응답엄도분별위7(2～11)개、11(9～14)개、4(2～6)개,3조감염자응답엄도적차이야유통계학의의(H=34.75,P＜0.01).3조감염자응답빈솔、응답강도화응답엄도종고도저의차위혈액감염＞10년조、혈액감염1～2년조、성접촉감염1～2년조.불동감염시간화불동감염도경적감염자대Nef태고화Gag태고적응답백분비화응답강도균고우기타태고.CD4계수＜200/μl、CD4계수위200～500/μl화≥500/μl3조감염자대17개태고적총응답강도범위분별위0～18 475 SFCs/106 PBMC、350～34 095 SFCs/106PBMC、490～21 550 SFCs/106 PBMC,단차이무통계학의의(H=2.93,P=0.23);3조감염자CTL응답엄도분별위3(0～8)개、10(2～17)개、10(1～17)개,3조간차이유통계학의의(H=14.72,P＜0.01),CD4계수＜200/μl적감염자CTL응답엄도저우CD4계수위200～500/μl화≥500/μl조.불동병독재량3조(＜LDL、LDL-1×104고패/ml화≥1×104고패/ml)표본대17개태고적총응답강도범위분별위490～18 475 SFCs/106PBMC、0～24 115 SFCs/106PBMC、770～34 095 SFCs/106 PBMC,단3조간차이무통계학의의(H=0.79,P=0.67);응답엄도분별위8(1～17)개、11(0～17)개、8(1～16)개,3조간차이야무통계학의의(H=5.27,P=0.07).결론 중국HIV/AIDS감염자중CTL응답다집중재Nef화Gag,저량개항원위HIV/AIDS감염적우세항원;감염시간화감염도경대CTL응답가산생현저영향;수감염시간적연장,면역반응적강도여응답비례균증가.저사신식대설계침대중국인군적AIDS역묘유교중요의의.
Objective To investigate and compare the features of the HIV-1-specific CTL responses among three HIV-infected groups with varied infection history. Methods Three HIV-infeeted groups were enrolled in this study, including two groups infected by blood transmission (one group has been infected for more than 10 years and the other for 1-2 years) and one group of the man who have sex with man. The HIV-1-specific CTL responses were quantified by an IFN-γ based ELISPot assay with a peptide matrix system containing overlapping peptides spanning the entire HIV-1 Clade B genomic consensus sequences. Results The responding rate of CTL responses against all 17 peptide pools among the group that infected 1-2 years,the group infected more than 10 years and the group of MSM were 40% ,65% ,23%. One way ANOVO analysis showed that the responding rate of CTL responses against all 17 peptide pools were statistical significant among the three groups (F=19.96, P＜0.01);the magnitude of CTL responses of the three groups were 0-5 835 SFCs/106 PBMC, 0-7 225 SFCs/106PBMC, 0-9 740SFCs/106pBMC, Kruskal-Wallis test showed that the magnitude of CTL responses were statistical significant among the three groups( H = 101.90 , P ＜0.01);the breadth of CTL were 7 ( 2-11 ), 11(9-14) and 4 (2-6) respectively and Kruskal- Wallis test showed that the breadth of CTL had no statistical significant among the three groups( H = 34. 75 ,P ＜0. 01 ). The sequence of responding rate, magnitude and breadth of CTL from high to low was the group that had been infected for more than 10 years, the group infected 1-2 years and the sex transmission group. The common characteristics of the CTL response among the three groups were that the responding rate and the magnitude of the peptide Nef and Gag was higher than other peptide's. The magnitude of CTL responses among three different CD4count groups (CD4 ＜ 200/μl, CD4 200-500/μl, CD4 ≥500/μl,) was 0-18 475 SFCs/106pBMC, 350-34 095 SFCs/106pBMC, 490-21 550 SFCs/106 PBMC and had no statistic difference among the three different CD4 groups(H=2.93, P=0.23) while the breadth of CTL was 3(0-8), 10(2-17), 10 (1-17)respoctively and the breadth of CTL was lower in the group of CD4 count less than 200/μl than the other two groups( H = 14. 72, P ＜ 0. 01 ). The magnitude of CTL responses among three different viral load (VL)groups (VL＜ LDL, LDL ＜ VL ＜ 1 × 104 copys/ml, VL≥1 ×104 copys/ml) was 490-18 475 SFCs/106pBMC, 0-24 115 SFCs/106pBMC, 770-34 095 SFCs/106 pBMC and had no statistic difference among the three different viral load groups ( H = 0.79, P=0.67) and the breadth of the three different viral load groups CTL was 8( 1-17), 11 (0-17), 8 (1-16) and Kruskal-Wallis test showed that there was no statistic difference among the three different viral load groups (H =5.27, P =0. 07). Conclusions All groups predominantly develope T cell immune responses against Nef and Gag proteins. With the elapse of HIV infection, the CTL responses are increased in both magnitude and responding rate. This information is important for vaccine development.
