CSISV人工感染SPF雏鸡和商品肉仔鸡后免疫器官的细胞凋亡
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摘要
鸡生长与免疫抑制综合征(暂定名,chicken stunt and immunosuppression syndrome,CSIS)是近年来发生的家禽生长和免疫抑制的重要传染性疾病之一。前期研究发现CSISV感染肉仔鸡后免疫器官存在凋亡现象,本试验用石蜡切片TUNEL法测定CSISV人工感染SPF雏鸡和肉仔鸡的免疫器官细胞凋亡指数,旨在研究CSISV引起免疫抑制的机理。
以感染病毒后72h的肉仔鸡法氏囊组织为研究对象,对石蜡切片TUNEL法检测细胞凋亡的试验条件进行优化,建立了检测细胞凋亡的TUNEL法。凋亡指数(AI)用每千个细胞中含有的凋亡细胞数表示,试验数据用mean±SD表示,SPSS13.0软件T检验分析组间差异性。用建立的石蜡切片TUNEL法测定了CSISV-ZZ2004毒株人工感染7日龄SPF雏鸡和商品肉仔鸡后免疫器官细胞凋亡情况,结果表明:
CSISV感染后,SPF雏鸡和商品肉仔鸡法氏囊、胸腺、脾脏的免疫细胞均发生大量凋亡。SPF雏鸡法氏囊细胞凋亡指数,在6～1056h极显著高于对照组(P<0.01),凋亡指数为15.532～39.386;胸腺细胞凋亡指数,在12h显著高于对照组(P<0.05),凋亡指数为6.262。24～1056h后极显著高于对照组(P<0.01),凋亡指数为7.719～23.956;脾脏细胞凋亡指数,在36h显著高于对照组(P<0.05),凋亡指数为3.841。48～1056h极显著高于对照组(P<0.01),凋亡指数为4.418～12.873。商品肉仔鸡免疫器官细胞凋亡指数曲线和SPF雏鸡基本一致,不同的是:细胞凋亡指数高峰的时间比SPF鸡晚48～72h。SPF鸡和商品肉仔鸡,法氏囊凋亡细胞在6～1056h髓质显著多于皮质(P<0.05);胸腺凋亡细胞在48～1056h皮质显著多于髓质(P<0.05);脾脏凋亡细胞在红髓和白髓都有,二者差异不显著(P>0.05)。说明CSISV感染后,法氏囊和胸腺中前B细胞和前T细胞大量凋亡,从而使能分化为成熟的淋巴细胞数量减少,使机体免疫功能下降,从免疫细胞的角度解释了CSIS发生免疫抑制的现象。
Recent years, chicken stunt and immunosupression syndrome (temporary name, CSIS) has been becoming one of important infectious diseases which break out on poultry. The disease’s main syndromes are growth stunt and immunosupression. The preceding research found apoptosis happened on immune organs after CSISV infection for broiler. The apoptosis index (AI) of immune organs specific pathogen free (SPF) chicks and broilers infected with CSISV is detected by using paraffin section TUNEL assay in this experiment. It is in order to research the immunosuppressive mechanism of CSISV.
It is carried out the research of broiler bursa of 72h after CSISV infection to optimize the experimental conditions of paraffin section TUNEL assay and establish the TUNEL assay to detect cell apoptosis. AI is expressed by the number of apoptotic cells per 1000 cells. The test data is expressed by mean±SD and the differences between control group and test group are analyzed by T test with SPSS13.0 software. Using the established TUNUL assay detect cell apoptosis of immune organs from 7-days-old SPF chicks and commercial broilers infected with CSISV-ZZ2004 strain. The results show:
The apoptosis appear in many immune cells of bursa、thymus and spleen from SPF chicks and commercial broilers after CSISV infection.The serious extent of apoptosis in the immune organs (including bursa, thymus and spleen) of SPF chicks infected with CSISV decrease respectively. The AI of bursa is 15.532～39.386 and very significant (P<0.01) higher than control group from 6h to 1056h after infection; The AI of thymus is 6.262 and significantly (P<0.05) higher than control group on 12h after infection. The AI of thymus is 7.719～23.956 and very significant (P<0.01) higher than control group from 24h to 1056h; The AI of spleen is 3.841 and significantly (P<0.05) higher than control group on 36h after infection. The AI of spleen is 4.418～12.873 and very significant (P<0.01) higher than control group from 48h to 1056h . The curves of AI of immune organs from commercial broilers infected with CSISV are almost similar with SPF chicks. However, there are a few differences on some aspects. The peak time of apoptosis is 48～72h later than SPF chicks. For SPF chicks and commercial broilers, the medulla has significant (P<0.05) more apoptotic cells than cortex in bursa from 6h to1056h; The cortex has significant (P<0.05) more apoptotic cells than medulla in thymus from 48h to1056h; The red pulp and white pulp all have apoptotic cells in spleen and the differences are not significant (P>0.05). These mean that apoptosis of lots of pre-T cells and pre-B cells in bursa and thymus appear after CSISV infection and the lymphocytes which can differentiate to mature lymphocytes are decreased. These make the immune function descend.This explain the immunosuppression of CSISV on immune cell aspect.
以感染病毒后72h的肉仔鸡法氏囊组织为研究对象,对石蜡切片TUNEL法检测细胞凋亡的试验条件进行优化,建立了检测细胞凋亡的TUNEL法。凋亡指数(AI)用每千个细胞中含有的凋亡细胞数表示,试验数据用mean±SD表示,SPSS13.0软件T检验分析组间差异性。用建立的石蜡切片TUNEL法测定了CSISV-ZZ2004毒株人工感染7日龄SPF雏鸡和商品肉仔鸡后免疫器官细胞凋亡情况,结果表明:
CSISV感染后,SPF雏鸡和商品肉仔鸡法氏囊、胸腺、脾脏的免疫细胞均发生大量凋亡。SPF雏鸡法氏囊细胞凋亡指数,在6～1056h极显著高于对照组(P<0.01),凋亡指数为15.532～39.386;胸腺细胞凋亡指数,在12h显著高于对照组(P<0.05),凋亡指数为6.262。24～1056h后极显著高于对照组(P<0.01),凋亡指数为7.719～23.956;脾脏细胞凋亡指数,在36h显著高于对照组(P<0.05),凋亡指数为3.841。48～1056h极显著高于对照组(P<0.01),凋亡指数为4.418～12.873。商品肉仔鸡免疫器官细胞凋亡指数曲线和SPF雏鸡基本一致,不同的是:细胞凋亡指数高峰的时间比SPF鸡晚48～72h。SPF鸡和商品肉仔鸡,法氏囊凋亡细胞在6～1056h髓质显著多于皮质(P<0.05);胸腺凋亡细胞在48～1056h皮质显著多于髓质(P<0.05);脾脏凋亡细胞在红髓和白髓都有,二者差异不显著(P>0.05)。说明CSISV感染后,法氏囊和胸腺中前B细胞和前T细胞大量凋亡,从而使能分化为成熟的淋巴细胞数量减少,使机体免疫功能下降,从免疫细胞的角度解释了CSIS发生免疫抑制的现象。
Recent years, chicken stunt and immunosupression syndrome (temporary name, CSIS) has been becoming one of important infectious diseases which break out on poultry. The disease’s main syndromes are growth stunt and immunosupression. The preceding research found apoptosis happened on immune organs after CSISV infection for broiler. The apoptosis index (AI) of immune organs specific pathogen free (SPF) chicks and broilers infected with CSISV is detected by using paraffin section TUNEL assay in this experiment. It is in order to research the immunosuppressive mechanism of CSISV.
It is carried out the research of broiler bursa of 72h after CSISV infection to optimize the experimental conditions of paraffin section TUNEL assay and establish the TUNEL assay to detect cell apoptosis. AI is expressed by the number of apoptotic cells per 1000 cells. The test data is expressed by mean±SD and the differences between control group and test group are analyzed by T test with SPSS13.0 software. Using the established TUNUL assay detect cell apoptosis of immune organs from 7-days-old SPF chicks and commercial broilers infected with CSISV-ZZ2004 strain. The results show:
The apoptosis appear in many immune cells of bursa、thymus and spleen from SPF chicks and commercial broilers after CSISV infection.The serious extent of apoptosis in the immune organs (including bursa, thymus and spleen) of SPF chicks infected with CSISV decrease respectively. The AI of bursa is 15.532～39.386 and very significant (P<0.01) higher than control group from 6h to 1056h after infection; The AI of thymus is 6.262 and significantly (P<0.05) higher than control group on 12h after infection. The AI of thymus is 7.719～23.956 and very significant (P<0.01) higher than control group from 24h to 1056h; The AI of spleen is 3.841 and significantly (P<0.05) higher than control group on 36h after infection. The AI of spleen is 4.418～12.873 and very significant (P<0.01) higher than control group from 48h to 1056h . The curves of AI of immune organs from commercial broilers infected with CSISV are almost similar with SPF chicks. However, there are a few differences on some aspects. The peak time of apoptosis is 48～72h later than SPF chicks. For SPF chicks and commercial broilers, the medulla has significant (P<0.05) more apoptotic cells than cortex in bursa from 6h to1056h; The cortex has significant (P<0.05) more apoptotic cells than medulla in thymus from 48h to1056h; The red pulp and white pulp all have apoptotic cells in spleen and the differences are not significant (P>0.05). These mean that apoptosis of lots of pre-T cells and pre-B cells in bursa and thymus appear after CSISV infection and the lymphocytes which can differentiate to mature lymphocytes are decreased. These make the immune function descend.This explain the immunosuppression of CSISV on immune cell aspect.
引文
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