Isotype-specific antibody-secreting cells in systemic and mucosal associated lymphoid tissues and antibody responses in serum of conventional pigs inoculated with PEDV.
Isotype-specific antibody-secreting cells in systemic and mucosal associated lymphoid tissues and antibody responses in serum of conventional pigs inoculated with PEDV.
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DOI：
10.1016/s0165-2427(01)00386-5
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发表时间：
2002-01-01
影响因子：
1.8
通讯作者：
Rubio P
中科院分区：
文献类型：
--
作者：
de Arriba ML;Carvajal A;Pozo J;Rubio P
本研究建立了一种酶联免疫斑点法（ELISPOT），用于检测猪流行性腹泻病毒（PEDV）感染后肠道相关淋巴组织（十二指肠、回肠固有层和肠系膜淋巴结）和全身组织（脾脏和血液）中的特异性抗体分泌细胞（ASC），以了解PEDV感染后产生的粘膜和全身抗体反应。用PEDV毒株CV-777的田间分离株口服接种总共28头11日龄的常规猪。在接种后第1 - 8天（PID），32%的猪出现腹泻，100%的猪出现病毒散毒。通过同种型ELISA分别从PID 12和15检测到针对PEDV的血清IgG和伊加抗体，在PID 32（IgG）和21（伊加）达到最大值。PEDV特异性IgM ASC发生在PID 4和7之间的所有组织中，其中肠固有层中的反应最强。从PID 21开始，肠道淋巴组织中伊加和IgG ASC反应明显，特异性ASC的最高数量对应于十二指肠固有层。在全身淋巴组织中，检测到的IgG和伊加ASC的数量低于粘膜组织，然而，在血液中，从PID 14直到实验结束，不断检测到伊加ASC的存在。还通过从肠系膜淋巴结、脾和血液分离的单核细胞（MNC）的二次体外刺激来研究对PEDV的记忆抗体应答。记忆B细胞应答在PID 21和25时显著，并且包括IgG和伊加ASC。据我们所知，这是研究PEDV感染后系统和肠道相关淋巴组织的不同位置中特异性ASC的存在和分布以及记忆B细胞的存在的第一份报告。
An enzyme-linked immunospot (ELISPOT) has been developed to detect porcine epidemic diarrhea virus (PEDV)-specific antibody secreting cells (ASC) in gut associated lymphoid tissues (duodenum and ileum lamina propria and mesenteric lymph nodes) and systemic locations (spleen and blood) of conventional pigs so as to characterise the mucosal and systemic antibody response generated by the infection with PEDV. A total number of 28 eleven-day-old conventional pigs were orally inoculated with the field isolate of the PEDV strain CV-777. Diarrhea was observed in 32% of the pigs and virus shedding was demonstrated in 100% between postinoculation day (PID) 1 and 8. Serum IgG and IgA antibodies to PEDV were detected by isotype ELISA from PID 12 and 15, respectively, reaching maximum values at PID 32 (IgG) and 21 (IgA). PEDV specific IgM ASC occurred in all the tissues between PID 4 and 7, with the strongest response in the intestinal lamina propria. IgA and IgG ASC responses were evident in the intestinal lymphoid tissues from PID 21, the highest number of specific ASC corresponded to the duodenum lamina propria. In the systemic lymphoid tissues the number of IgG and IgA ASC detected were lower than in the mucosal tissues, however, in the blood, presence of IgA ASC was constantly detected from PID 14 until the end of the experiment. Memory antibody response to the PEDV was also studied by secondary in vitro stimulation of the mononuclear cells (MNC) isolated from mesenteric lymph nodes, spleen and blood. The memory B cell response was prominent at PID 21 and 25 and consisted in IgG and IgA ASC. To our knowledge, this is the first report to research into the presence and distribution of specific ASC in different locations of the systemic and the gut associated lymphoid tissues after a PEDV infection as well as the presence of memory B cells.
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影响因子：
2.6
作者：
Carvajal A;Lanza I;Diego R;Rubio P;Cármenes P
通讯作者：
Cármenes P
影响因子：
2.2
作者：
Berthon P;Bernard S;Salmon H;Binns RM
通讯作者：
Binns RM
影响因子：
2.2
作者：
CZERKINSKY, CC;NILSSON, LA;TARKOWSKI, A
通讯作者：
TARKOWSKI, A
影响因子：
2.2
作者：
SEDGWICK, JD;HOLT, PG
通讯作者：
HOLT, PG
影响因子：
1.8
作者：
Saif, LJ
通讯作者：
Saif, LJ