Wiltrout R H, Mathieson B J, Talmadge J E, Reynolds C W, Zhang S R, Herberman R B, Ortaldo J R
J Exp Med. 1984 Nov 1;160(5):1431-49. doi: 10.1084/jem.160.5.1431.
Natural killer (NK) activity in the rat and human has been attributed to cells having the morphology of large granular lymphocytes (LGL). However, this association has been less clear in the mouse, largely because of difficulties in obtaining highly enriched populations of LGL from normal spleen and blood. We have previously observed that the administration of the biological response modifier (BRM) maleic anhydride divinyl ether (MVE-2) strongly augmented NK activity in lung and liver, and the augmented NK activity coincided with increased resistance to the formation of experimental metastases in these organs. The degree of NK augmentation was most striking in the liver, an unexpected and previously unreported observation. In the present study, both MVE-2 or Corynebacterium parvum induced a dramatic augmentation of liver NK activity, which reached maximum levels 3-5 d after treatment. This augmentation of NK activity in the liver coincided with a large increase in the number of lymphoid cells with the morphological characteristics of LGL that could be isolated from enzymatically digested suspensions of perfused liver. The yield of LGL per liver following BRM treatment corresponded to a 10-50-fold increase as compared to normal mice. LGL were purified from these enzymatically digested suspensions of perfused liver by depletion of adherent cells on nylon wool columns and subsequent enrichment for low-density lymphoid cells by fractionation on Percoll density gradients. The enrichment of LGL correlated with greatly increased NK activity against YAC-1. Conversely, the higher-density fractions were depleted of both LGL and NK activity. This increase in NK activity in the liver was suppressed by in vivo treatment with anti-asialo GM1 (asGM1) serum. This treatment also resulted in a corresponding reduction in both the total number and percentage of LGL. By flow cytometry analysis, the phenotype of the majority of these highly cytolytic LGL isolated from the livers of BRM-treated mice were asGM1+, Thy-1+, Ly-5+, Qa-5+, Mac-1+, and Gma-1+, whereas these LGL were Ly-1-, Lyt-2-, L3T4-, and surface Ig-. We conclude that the livers of BRM-treated mice can provide a rich source of highly active mouse LGL that could be used for further characterization of this lymphocyte subset. Further, these studies imply a potential for BRM therapy of neoplastic or viral diseases through augmentation of organ-associated immune responses.
大鼠和人类的自然杀伤（NK）活性一直被认为与具有大颗粒淋巴细胞（LGL）形态的细胞有关。然而，在小鼠中这种关联并不那么明确，主要是因为从正常脾脏和血液中获得高度富集的LGL群体存在困难。我们之前观察到，给予生物反应调节剂（BRM）马来酸二乙烯醚（MVE-2）可强烈增强肺和肝脏中的NK活性，而增强的NK活性与这些器官中对实验性转移形成的抵抗力增加相一致。NK增强程度在肝脏中最为显著，这是一个意外且先前未报道的观察结果。在本研究中，MVE-2或短小棒状杆菌均能显著增强肝脏NK活性，在治疗后3 - 5天达到最高水平。肝脏中NK活性的这种增强与从灌注肝脏的酶消化悬液中分离出的具有LGL形态特征的淋巴细胞数量大幅增加相一致。与正常小鼠相比，BRM处理后每只肝脏中LGL的产量增加了10 - 50倍。通过在尼龙毛柱上去除贴壁细胞，随后在Percoll密度梯度上进行分级分离以富集低密度淋巴细胞，从这些灌注肝脏的酶消化悬液中纯化LGL。LGL的富集与针对YAC-1的NK活性大幅增加相关。相反，较高密度的组分中LGL和NK活性均减少。肝脏中NK活性的这种增加可被体内注射抗唾液酸GM1（asGM1）血清所抑制。这种处理还导致LGL的总数和百分比相应减少。通过流式细胞术分析，从BRM处理小鼠肝脏中分离出的大多数高细胞毒性LGL的表型为asGM1 +、Thy-1 +、Ly-5 +、Qa-5 +、Mac-1 +和Gma-1 +，而这些LGL为Ly-1 -、Lyt-2 -、L3T4 -且表面无免疫球蛋白。我们得出结论，BRM处理小鼠的肝脏可提供丰富的高活性小鼠LGL来源，可用于进一步表征该淋巴细胞亚群。此外，这些研究暗示了BRM通过增强器官相关免疫反应治疗肿瘤或病毒性疾病的潜力。
