雏鸡感染鸡传染性贫血病毒后免疫变化和对新城疫疫苗的应答及其免疫机理
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摘要
本项研究采用免疫组织化学、免疫胶体金及免疫酶方法，首次对CIAV感染1日龄雏鸡和CIAV感染雏鸡ND疫苗免疫及vNDV攻击后，其外周血液的T细胞、B细胞数量和IgG、IgM、IgA含量；中枢免疫器官胸腺和法氏囊，外周免疫器官脾脏及呼吸道和消化道局部粘膜免疫组织哈德尔腺和盲肠扁桃体的ANAE~+、CD_4~+、CD_8~+T细胞、IgG、IgM、IgA抗体生成细胞数量和CD_4~+与CD_8~+T细胞比值以及泪液、气管液、肠液、胆汁的IgA、IgM、IgG含量的动态变化规律和CIAV感染对ND疫苗的免疫保护率等进行了全面系统地研究。旨在深入系统地揭示CIAV的免疫致病机理和其对ND疫苗免疫应答的影响及其机理，为CIAV的综合防制提供重要的科学依据。研究结果发现：
1、1日龄雏鸡感染CIAV后28天内，其外周血液T细胞、B细胞数量和IgG、IgM、IgA含量均明显低于未感染CIAV的对照雏鸡，表明CIAV感染雏鸡外周血液的细胞和体液免疫机能都下降，这与CIAV感染后免疫器官组织遭受严重破坏密切相关。
2、1日龄雏鸡感染CIAV后35天内，免疫器官胸腺、法氏囊、脾脏的CD_4~+、CD_8~+、ANAE~+T细胞和IgG、IgM、IgA抗体生成细胞数量均程度不同地较未感染CIAV的对照雏鸡明显减少或降低，特别是CD_4~+与CD_8~+T细胞比值，感染雏鸡明显低于对照雏鸡，提示CIAV感染雏鸡免疫器官，不仅细胞免疫机能抑制，而且体液免疫机能也明显受到影响，免疫器官的免疫机能下降，是导致感染雏鸡免疫机能低下的主导环节。
3、1日龄雏鸡感染CIAV后28天内，哈德尔腺和盲肠扁桃体的ANAE~+及CD_4~+、CD_8~+T细胞和IgA、IgM、IgG抗体生成细胞数量以及泪液、气管液、肠液、胆汁的IgA、IgM、IgG含量均明显低于对照雏鸡，表明CIAV感染雏鸡后，其呼吸道和消化道局部粘膜的细胞免疫和体液免疫机能均下降，这与CIAV感染雏鸡对其它某些病原易感性提高密切相关。
4、CIAV感染雏鸡ND免疫后28天内，其外周血液的IgG、IgM、IgA含量和刚抗体滴度，均明显低于未感染CIAV的ND免疫对照雏鸡，提示CIAV感染雏鸡，外周血液对ND疫苗的体液免疫应答机能降低。
5、CIAV感染雏鸡ND免疫后28天内，胸腺、法氏囊、脾脏等免疫器官的CD_4~+、CD_8~+T细胞和IgG、IgM、IgA抗体生成细胞数量及CD_4~+与CD_8~+T细胞比值，均明
博士学位论文 中文摘要 东北农业大学
显低于未感染CIAV的ND免疫雏鸡，表明CIAV感染雏鸡免疫器官对ND疫苗的免
疫应答机能下降。
6、CIAV感染雏鸡ND免疫后28天内，哈德尔腺、盲肠扁桃体以及泪液、气
管液、肠液、胆汁的上述各项兔疫学指标，均不同程度的明显低于未感染CIAV的
ND免疫雏鸡，表明CIAV感染雏鸡呼吸道和消化道局部粘膜对ND疫苗的细胞和体
液免疫应答机能下降。
7、CIAV感染ND兔疫雏鸡VNDV攻击后，其兔疫保护率（60％）明显低于未
感染CIAV的ND免疫对照雏鸡（100％），这与CIAV感染导致雏鸡免疫机能抑制，
对ND疫苗的免疫应答机能降低密切相关。
上述研究结果，首次全面系统地揭示了CIAV感染雏鸡外周血液、全身免疫器
官、消化道和呼吸道局部粘膜的细胞免疫和体液免疫均降低，并阐述CIAV感染雏
鸡的细胞免疫与体液兔疫变化的内在联系和变化规律；同时，还揭示了CIAV感染
雏鸡对ND疫苗免疫的外周血液、兔疫器官、局部粘膜的细胞免疫和体液兔疫应答
减弱；CIAV感染ND免疫雏鸡VNDV攻击后，其死亡率明显提高与CIAV感染雏鸡
免疫器官组织的细胞免疫和体液兔疫机能受损，使其对ND疫茵兔疫的应答机能降
低有关。
The dynamic changes of the number of T or B lymphocytes and the content of immunoglobulin (Ig) G, IgM, IgA in peripheral blood, the number of CD4+, CD8+ lymphocyte subpopulation , TANAE+ cells and IgG, IgM, IgA antibody-producing cells, the ratios of CD4+ and CDg+ T cells in immune organs (thymus, bursa of Fabricius and spleen) and immune tissues (cecal tonsils and Harderian glands), the content of IgA, IgM, IgG in local fluid (tear, tracheal fluid, intestinal fluid and bile) of one-day chicks infected with chicken infectious anemia virus (CIAV) and CIAV-infected chicks immunized with Newcastle disease (ND) vaccine were determined respectively by immunohistochemistry, immunogold-silves and immunoenzyme staining in order to reveal mechanism of immunopathogenesis of chicks infected with CIAV and influence of immune responses of CIAV-infected chicks to ND vaccine. The results were as follows:
1. The numbers of T,B lymphocytes and the content of IgG, IgM, IgA in peripheral blood of chicks infected with CIAV were significantly decreased compared with controls 7-28 days postinfection (PI). The results showed that cellular and humoral immune function in peripheral blood of chicks infected with CIAV were lowered.
2. The number of CD4+, CDg+ lymphocyte subpopulation , ANAE+ T cells and IgG, IgM, IgA antibody-producing cells in immune organs (thymus, bursa of Fabricius and spleen) of chicks infected with CIAV were significantly decreased compared with control 7~35 days PI. Especially, the ratios of CD4+ and CD8+ T cells of CIAV-infected chicks were markedly lowered compared with chicks no-infected with CIAV. The results indicated that the cellular immune function in immune organs of CIAV-infected chicks not only was repressed but humoral immune function was lowered.
3. The number of CD4+ and CD8+ lymphocytes subpopulation , ANAE+ T cells and IgG, IgM, IgA antibody-producing cells in the local immune tissues (cecal tonsils and Harderian glands), the content of IgA, IgM, IgG in the local fluid (tear, tracheal fluid, intestinal fluid and bile) of chicks infected with CIAV were significantly decreased compared with controls 7-28 days PI. The results showed that the both cellular and
humoral immune function of local mucous membrane in the digestive and respiratory duct of CIAV-infected chicks was lowered.
4. The number of CD/ lymphocytes subpopulation in the immune organs and tissues of CIAV-infected chicks decreased lower than CD8+ T cells. The results indicated that CIAV infection destroyed CD4+ T-cell subsets more serious than CDg+.
5. The content of IgG, IgM, IgA and HI titer in serum were significantly lower in infected-vaccinated chicks than vaccinated control chicks 7-28 days postvaccination (PV). These indicated that humoral immune response in peripheral blood of CIAV-infected chicks to ND vaccine were markedly lowered.
6. The number of CD4+ and CDg+ lymphocytes subpopulation and IgA, IgM, IgG antibody-producing cells , the ratios of CD4+ and CDg+ T-cell subsets in immune organs (thymus, bursa of Fabricius and spleen) of CIAV-infected chicks vaccinated ND vaccine were significantly decreased compared with vaccinated control chicks 7-28 days PV. The results showed that immune responses in immune organs of chicks infected with CIAV to ND vaccine were lowered.
7. The number of CD4+ and CD8+ lymphocytes subpopulation and IgA, IgM, IgG antibody-producing cells, the ratios of CD4+ and CD8+ T-cell in the cecal tonsils and Harderian glands, the content of IgA, IgM, IgG in tear, tracheal fluid, intestinal fluid and bile of CIAV-infected chicks vaccinated with ND vaccine were significantly decreased compared with vaccinated control chicks 7-28 days PV. The results showed that immune responses of local mucous membrane of CIAV-infected chicks to ND vaccine were lowered.
8. The immune protective rates of CIAV-infected chicks vaccinated with ND vaccine (60%) were markedly lower than that of the ND-vaccinated control birds (100%) after challenge with vND
1、1日龄雏鸡感染CIAV后28天内，其外周血液T细胞、B细胞数量和IgG、IgM、IgA含量均明显低于未感染CIAV的对照雏鸡，表明CIAV感染雏鸡外周血液的细胞和体液免疫机能都下降，这与CIAV感染后免疫器官组织遭受严重破坏密切相关。
2、1日龄雏鸡感染CIAV后35天内，免疫器官胸腺、法氏囊、脾脏的CD_4~+、CD_8~+、ANAE~+T细胞和IgG、IgM、IgA抗体生成细胞数量均程度不同地较未感染CIAV的对照雏鸡明显减少或降低，特别是CD_4~+与CD_8~+T细胞比值，感染雏鸡明显低于对照雏鸡，提示CIAV感染雏鸡免疫器官，不仅细胞免疫机能抑制，而且体液免疫机能也明显受到影响，免疫器官的免疫机能下降，是导致感染雏鸡免疫机能低下的主导环节。
3、1日龄雏鸡感染CIAV后28天内，哈德尔腺和盲肠扁桃体的ANAE~+及CD_4~+、CD_8~+T细胞和IgA、IgM、IgG抗体生成细胞数量以及泪液、气管液、肠液、胆汁的IgA、IgM、IgG含量均明显低于对照雏鸡，表明CIAV感染雏鸡后，其呼吸道和消化道局部粘膜的细胞免疫和体液免疫机能均下降，这与CIAV感染雏鸡对其它某些病原易感性提高密切相关。
4、CIAV感染雏鸡ND免疫后28天内，其外周血液的IgG、IgM、IgA含量和刚抗体滴度，均明显低于未感染CIAV的ND免疫对照雏鸡，提示CIAV感染雏鸡，外周血液对ND疫苗的体液免疫应答机能降低。
5、CIAV感染雏鸡ND免疫后28天内，胸腺、法氏囊、脾脏等免疫器官的CD_4~+、CD_8~+T细胞和IgG、IgM、IgA抗体生成细胞数量及CD_4~+与CD_8~+T细胞比值，均明
博士学位论文 中文摘要 东北农业大学
显低于未感染CIAV的ND免疫雏鸡，表明CIAV感染雏鸡免疫器官对ND疫苗的免
疫应答机能下降。
6、CIAV感染雏鸡ND免疫后28天内，哈德尔腺、盲肠扁桃体以及泪液、气
管液、肠液、胆汁的上述各项兔疫学指标，均不同程度的明显低于未感染CIAV的
ND免疫雏鸡，表明CIAV感染雏鸡呼吸道和消化道局部粘膜对ND疫苗的细胞和体
液免疫应答机能下降。
7、CIAV感染ND兔疫雏鸡VNDV攻击后，其兔疫保护率（60％）明显低于未
感染CIAV的ND免疫对照雏鸡（100％），这与CIAV感染导致雏鸡免疫机能抑制，
对ND疫苗的免疫应答机能降低密切相关。
上述研究结果，首次全面系统地揭示了CIAV感染雏鸡外周血液、全身免疫器
官、消化道和呼吸道局部粘膜的细胞免疫和体液免疫均降低，并阐述CIAV感染雏
鸡的细胞免疫与体液兔疫变化的内在联系和变化规律；同时，还揭示了CIAV感染
雏鸡对ND疫苗免疫的外周血液、兔疫器官、局部粘膜的细胞免疫和体液兔疫应答
减弱；CIAV感染ND免疫雏鸡VNDV攻击后，其死亡率明显提高与CIAV感染雏鸡
免疫器官组织的细胞免疫和体液兔疫机能受损，使其对ND疫茵兔疫的应答机能降
低有关。
The dynamic changes of the number of T or B lymphocytes and the content of immunoglobulin (Ig) G, IgM, IgA in peripheral blood, the number of CD4+, CD8+ lymphocyte subpopulation , TANAE+ cells and IgG, IgM, IgA antibody-producing cells, the ratios of CD4+ and CDg+ T cells in immune organs (thymus, bursa of Fabricius and spleen) and immune tissues (cecal tonsils and Harderian glands), the content of IgA, IgM, IgG in local fluid (tear, tracheal fluid, intestinal fluid and bile) of one-day chicks infected with chicken infectious anemia virus (CIAV) and CIAV-infected chicks immunized with Newcastle disease (ND) vaccine were determined respectively by immunohistochemistry, immunogold-silves and immunoenzyme staining in order to reveal mechanism of immunopathogenesis of chicks infected with CIAV and influence of immune responses of CIAV-infected chicks to ND vaccine. The results were as follows:
1. The numbers of T,B lymphocytes and the content of IgG, IgM, IgA in peripheral blood of chicks infected with CIAV were significantly decreased compared with controls 7-28 days postinfection (PI). The results showed that cellular and humoral immune function in peripheral blood of chicks infected with CIAV were lowered.
2. The number of CD4+, CDg+ lymphocyte subpopulation , ANAE+ T cells and IgG, IgM, IgA antibody-producing cells in immune organs (thymus, bursa of Fabricius and spleen) of chicks infected with CIAV were significantly decreased compared with control 7~35 days PI. Especially, the ratios of CD4+ and CD8+ T cells of CIAV-infected chicks were markedly lowered compared with chicks no-infected with CIAV. The results indicated that the cellular immune function in immune organs of CIAV-infected chicks not only was repressed but humoral immune function was lowered.
3. The number of CD4+ and CD8+ lymphocytes subpopulation , ANAE+ T cells and IgG, IgM, IgA antibody-producing cells in the local immune tissues (cecal tonsils and Harderian glands), the content of IgA, IgM, IgG in the local fluid (tear, tracheal fluid, intestinal fluid and bile) of chicks infected with CIAV were significantly decreased compared with controls 7-28 days PI. The results showed that the both cellular and
humoral immune function of local mucous membrane in the digestive and respiratory duct of CIAV-infected chicks was lowered.
4. The number of CD/ lymphocytes subpopulation in the immune organs and tissues of CIAV-infected chicks decreased lower than CD8+ T cells. The results indicated that CIAV infection destroyed CD4+ T-cell subsets more serious than CDg+.
5. The content of IgG, IgM, IgA and HI titer in serum were significantly lower in infected-vaccinated chicks than vaccinated control chicks 7-28 days postvaccination (PV). These indicated that humoral immune response in peripheral blood of CIAV-infected chicks to ND vaccine were markedly lowered.
6. The number of CD4+ and CDg+ lymphocytes subpopulation and IgA, IgM, IgG antibody-producing cells , the ratios of CD4+ and CDg+ T-cell subsets in immune organs (thymus, bursa of Fabricius and spleen) of CIAV-infected chicks vaccinated ND vaccine were significantly decreased compared with vaccinated control chicks 7-28 days PV. The results showed that immune responses in immune organs of chicks infected with CIAV to ND vaccine were lowered.
7. The number of CD4+ and CD8+ lymphocytes subpopulation and IgA, IgM, IgG antibody-producing cells, the ratios of CD4+ and CD8+ T-cell in the cecal tonsils and Harderian glands, the content of IgA, IgM, IgG in tear, tracheal fluid, intestinal fluid and bile of CIAV-infected chicks vaccinated with ND vaccine were significantly decreased compared with vaccinated control chicks 7-28 days PV. The results showed that immune responses of local mucous membrane of CIAV-infected chicks to ND vaccine were lowered.
8. The immune protective rates of CIAV-infected chicks vaccinated with ND vaccine (60%) were markedly lower than that of the ND-vaccinated control birds (100%) after challenge with vND
引文
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