Iglesias G, Pijoan C, Molitor T
Facultad de Estudios Superiores, Cuautitlan UNAM, Cuautitlan Izcalli, Mexico.
Comp Immunol Microbiol Infect Dis. 1992 Oct;15(4):249-59. doi: 10.1016/0147-9571(92)90004-b.
Alveolar macrophages (AM) infected with Pseudorabies virus (PRV) were compared to noninfected AM for cytotoxicity against foreign or transformed cells and production of interferon (IFN). Five PRV strains were used to infect AM including strains that are known to be highly virulent for pigs, i.e. strain 4892 and strain S-62 as well as strains that are regarded as mild or nonvirulent, i.e. BUK and Bartha. The multiplicity of infection ranged from 0.005 to 0.05 TCID50/cell. The target cells in the cytotoxicity assays were either chicken red blood cells, PRV-infected vero cells, or human myeloblastoma cells (K562 cell line). For the production of IFN, AM cultures were treated with polyinosinic:polycytidylic acid (Poly I:C) diluted in tissue culture media at a concentration of 5 micrograms/10(6) cells. Culture supernatants were collected at various times poststimulation and tested for antiviral activity using the Vesicular Stomatitis Virus replication inhibition test. Swine AM were able to lyse chicken red blood cells in an antibody-independent way but not in an antibody-dependent way, whereas lysis of PRV-infected vero cells was accomplished both ways. The cytotoxicity against chicken red blood cells was reduced in the PRV-infected AM as compared to noninfected cells, particularly in AM infected with virulent PRV strains. Specific 51Cr release values for AM infected with S-62 and 4892 strains were 14 and 19, while the noninfected AM had values of 36. Similarly, in the antibody-dependent cytotoxicity assay against PRV-infected vero cells there was no activity of AM against K562 cells. The production of IFN was readily stimulated with Poly I:C. The optimal time for supernatant collection was between 12 and 16 h poststimulation. The antiviral activity was abrogated by treatment of the supernatant with antiserum against human leukocyte IFN; it was therefore considered to be due to interferon-alpha (IFN alpha) released from the macrophages. The antiviral activity present in supernatants of PRV-infected AM was reduced compared to noninfected AM. The difference between AM cultures infected with virulent strains of PRV and noninfected AM cultures was statistically significant at P < or = 0.025. The results provide support to the premise that the role of AM in lung defense can be compromised by PRV infection.
将感染伪狂犬病病毒(PRV)的肺泡巨噬细胞(AM)与未感染的AM进行比较,观察其对外来细胞或转化细胞的细胞毒性以及干扰素(IFN)的产生情况。使用5种PRV毒株感染AM,其中包括已知对猪具有高毒力的毒株,即4892株和S - 62株,以及被视为温和或无毒的毒株,即BUK株和Bartha株。感染复数范围为0.005至0.05 TCID50/细胞。细胞毒性试验中的靶细胞为鸡红细胞、PRV感染的非洲绿猴肾细胞(vero细胞)或人成髓细胞瘤细胞(K562细胞系)。对于IFN的产生,将AM培养物用在组织培养基中稀释至浓度为5微克/10⁶细胞的聚肌苷酸:聚胞苷酸(Poly I:C)处理。在刺激后的不同时间收集培养上清液,并使用水疱性口炎病毒复制抑制试验检测抗病毒活性。猪AM能够以非抗体依赖的方式裂解鸡红细胞,但不能以抗体依赖的方式裂解;而PRV感染的vero细胞的裂解两种方式均可实现。与未感染细胞相比,PRV感染的AM对鸡红细胞的细胞毒性降低,尤其是感染强毒PRV毒株的AM。感染S - 62株和4892株的AM的特异性⁵¹Cr释放值分别为14和19,而未感染的AM的值为36。同样,在针对PRV感染的vero细胞的抗体依赖细胞毒性试验中,AM对K562细胞无活性。Poly I:C能轻易刺激IFN的产生。收集上清液的最佳时间为刺激后12至16小时。用抗人白细胞IFN抗血清处理上清液后,抗病毒活性被消除;因此认为其是由巨噬细胞释放的α干扰素(IFNα)所致。与未感染的AM相比,PRV感染的AM上清液中的抗病毒活性降低。感染强毒PRV毒株的AM培养物与未感染的AM培养物之间的差异在P≤0.025时具有统计学意义。这些结果支持了PRV感染可能损害AM在肺部防御中作用的前提。