Time course of the porcine cellular and humoral immune responses in vivo against pseudorabies virus after inoculation and challenge: Significance of in vitro antigenic restimulation
Time course of the porcine cellular and humoral immune responses in vivo against pseudorabies virus after inoculation and challenge: Significance of in vitro antigenic restimulation
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DOI：
10.1016/s0165-2427(98)00175-5
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发表时间：
1998-09-16
影响因子：
1.8
通讯作者：
Bianchi, ATJ
中科院分区：
文献类型：
--
作者：
De Bruin, MGM;De Visser, YE;Bianchi, ATJ
研究了伪狂犬病病毒（PRV）gE（-）突变株M141免疫猪后，用野生型病毒NIA-3攻毒后，猪的细胞免疫和体液免疫应答的时间进程。从血液样品中分离外周血单核细胞（PBMC）;一半直接使用，一半在体外用PRV再刺激，然后用于细胞溶解试验。我们确定了PRV特异性淋巴细胞增殖和细胞溶解反应的时间进程和程度。此外，还检查了血清样本中的中和抗体。接种后，用FACScan测定外周血中各种淋巴细胞亚群的频率。接种后一周，T淋巴细胞大量增殖，并观察到B淋巴细胞反应。当直接使用PBMC而不进行再刺激时，仅15%的PRV感染的靶细胞被裂解，并且约15-20%的未感染的靶细胞被裂解。相反，当用PRV再刺激PBMC时，高达50%的PRV感染的靶细胞被裂解，而仅30%的未感染的靶细胞被裂解。接种后，循环中各种T淋巴细胞亚群的频率没有明显变化，这表明循环中PRV特异性淋巴细胞的数量很少。攻毒后，T淋巴细胞反应增强，但B淋巴细胞反应没有增强。当直接使用PBMC时，只有20%的PRV感染和未感染的靶细胞在攻击后裂解。相反，当PBMC用PRV再刺激时，它们再次比未感染的靶细胞裂解更多的PRV感染的靶细胞。攻毒后比接种后更长时间内检测到溶细胞细胞。由于只有在淋巴细胞用PRV再刺激后才能清楚地检测到细胞溶解，可能是它们不优先定位于血液中，或者它们在血液中太少而不能在体外进一步抗原再刺激的情况下检测到。这些淋巴细胞可以替代地定位在其他组织中，例如粘膜组织、扁桃体和引流淋巴结。这种PRV特异性细胞溶解细胞的储存库是否对清除病毒很重要仍然是未知的。在这项研究中，我们证明了PRV特异性淋巴细胞在循环后，他们在体外再刺激PRV。(C)1998 Eisevier Science B. V.保留所有权利。
We investigated the time course of porcine cellular and humoral immune responses against pseudorabies virus (PRV) after pigs were inoculated with PRV gE(-) mutant strain M141 and challenged with wild-type virus NIA-3. Peripheral blood mononuclear cells (PBMC) were isolated from blood samples; half were used directly and half were restimulated with PRV in vitro before use in a cytolytic assay. We determined time course and extent of PRV-specific lymphoproliferative and cytolytic response. In addition, serum samples were examined for neutralizing antibodies. After inoculation, the frequency of various lymphocyte subsets in peripheral blood was determined by FACScan. One week after inoculation, T-lymphocytes proliferated abundantly and a B-lymphocyte response was observed. When PBMC were used directly without restimulation, only 15% of the PRV-infected target cells were lysed, and about 15-20% of uninfected target cells were lysed. In contrast, when PBMC were restimulated with PRV, up to 50% of the PRV-infected target cells were lysed while only 30% of the uninfected target cells were lysed. The frequency of various T-lymphocyte subsets in the circulation did not change significantly after inoculation, which indicates that the number of PRV-specific lymphocytes in circulation was very small. After challenge, the T-lymphocyte response was enhanced, but the B-lymphocyte response was not. When PBMC were used directly, only 20% of the PRV-infected and uninfected target cells were lysed after challenge. In contrast, when PBMC were restimulated with PRV, they again lysed more PRV-infected target cells than uninfected target cells. Cytolytic cells were detected for a longer period after challenge than after inoculation. Since it was only possible to clearly detect cytolysis after lymphocytes were restimulated with PRV, it may be that they do not preferentially localize in blood or that they are too few in blood to be detected without further antigenic restimulation in vitro. These lymphocytes may instead localize in other tissues, such as mucosal tissues, tonsils and draining lymph nodes. Whether such a reservoir of PRV-specific cytolytic cells is important in clearing the virus is still unknown. In this study we demonstrated PRV-specific lymphocytes in circulation after they were restimulated in vitro with PRV. (C) 1998 EIsevier Science B.V. All rights reserved.