Lohse Louise, Nielsen Jens, Eriksen Lis
Danish Institute for Food and Veterinary Research, Kalvehave, Denmark.
APMIS. 2006 Jan;114(1):23-31. doi: 10.1111/j.1600-0463.2006.apm_301.x.
In vivo depletion of lymphocyte subsets allows investigation of the role of specific subsets in protective immunity. In the present study we evaluated the effects of long-term, low-dose treatment with murine monoclonal antibodies (mAbs) against porcine CD4 and CD8 surface antigens on lymphocyte subsets in pigs. Four-week-old pigs were treated by intramuscular injections of hybridoma cell culture supernatants containing anti-CD mAbs twice a week for a period of 5 weeks. The immunomodulatory effects of the treatments were assessed by flow cytometry (FCM) analysis of peripheral blood lymphocytes. Treatment with the anti-CD4 mAb almost completely eliminated the CD4(+) T-cell subset from the circulation after 2 weeks of therapy. This depletion persisted until the end of the experimental period 5 weeks after initiated therapy. Treatment with the anti-CD8 mAb was less effective, reducing the CD8(+) T-cell subset in peripheral blood by approximately 50% of the initial level after 3 weeks of therapy. Further, the anti-CD8 mAb-treated pigs showed a parallel increase in the CD4(+) T-cell subset from day 7. Two-colour FCM analysis indicated that a shift in phenotype from single-positive CD4(+)/CD8(-) to double-positive CD4(+)/CD8(+) T-cells might have occurred in these pigs. In the present experiment we demonstrated specific modulation of the peripheral blood T-lymphocyte population in pigs with continuous low-dose injections of specific mAb. The ability to modulate individual T-cell subsets should provide a method to elucidate their functionality in protection against infectious disease.
体内淋巴细胞亚群的耗竭有助于研究特定亚群在保护性免疫中的作用。在本研究中,我们评估了用抗猪CD4和CD8表面抗原的鼠单克隆抗体(mAb)进行长期、低剂量治疗对猪淋巴细胞亚群的影响。对4周龄的猪每周两次肌肉注射含有抗CD mAb的杂交瘤细胞培养上清液,持续5周。通过对外周血淋巴细胞进行流式细胞术(FCM)分析来评估治疗的免疫调节作用。用抗CD4 mAb治疗2周后,循环中的CD4(+) T细胞亚群几乎完全消失。这种耗竭一直持续到治疗开始后5周的实验期结束。用抗CD8 mAb治疗效果较差,治疗3周后外周血中CD8(+) T细胞亚群减少至初始水平的约50%。此外,从第7天起,用抗CD8 mAb治疗的猪的CD4(+) T细胞亚群平行增加。双色FCM分析表明,这些猪可能发生了表型从单阳性CD4(+)/CD8(-) T细胞向双阳性CD4(+)/CD8(+) T细胞的转变。在本实验中,我们证明了通过持续低剂量注射特异性mAb对猪外周血T淋巴细胞群体进行特异性调节。调节单个T细胞亚群的能力应能提供一种方法来阐明它们在预防传染病中的功能。