Archana Archana, Patel Prem Shankar, Kumar Rajesh, Nath Gopal
Department of Microbiology, Institute of Medical Sciences, Banaras Hindu University, Varanasi, 221005 India.
Department of Nephrology, Institute of Medical Sciences, Banaras Hindu University, Varanasi, 221005 India.
Virusdisease. 2021 Mar;32(1):38-45. doi: 10.1007/s13337-021-00673-8. Epub 2021 Mar 10.
Bacteriophage therapy is currently experiencing a renaissance. Therapeutic efficacy of bacteriophages depends on phage-bacterial and phage-host interactions. The appearance of neutralizing anti-phage antibody has been speculated to be one of the few reasons for bacteriophage therapy's failure. This study aimed to know whether there is a rise in the neutralizing antibody on the parenteral injection of bacteriophages in an animal model. This study included bacteriophages against five different bacteria, namely , , , and These bacteriophages were isolated, propagated and purified. Bacteriophage specificity was confirmed by spot testing on the respective bacterial lawn. Weekly subcutaneous injection of purified bacteriophages (10PFU) was given to five rabbits for six weeks. Blood samples were collected before administering the next dose every week. The antibody response was tested by phage neutralization followed by plaque assay by using double agar overlay method. The rise in anti-phage neutralizing antibodies was observed usually after the 3 week after immunization. Complete neutralization of bacteriophages could be seen between 3 and 5 weeks after immunization. A further rise in bacteriophage counts (PFU), especially on 1:1000 and 1:2000 serum dilutions, could be noticed by the end of 6th week against most bacteriophages injected. Background anti-phage neutralizing antibodies were observed against bacteriophage specific to However, it was absent against bacteriophages specific to other four bacteria. Bacteriophage interacts with mammalian host and induces anti-phages neutralizing antibody production. However, neutralization of phage depends on repeated administration and duration of therapy. The significant rise in neutralizing antibody could be seen at the end of 3rd week. Therefore, bacteriophage can be effectively used in acute cases where therapy duration is less than 2 weeks. However, for prolonged therapy, bacteriophage cocktail of different antigenicity may be suggested.
噬菌体疗法目前正在复兴。噬菌体的治疗效果取决于噬菌体与细菌以及噬菌体与宿主之间的相互作用。据推测，中和性抗噬菌体抗体的出现是噬菌体疗法失败的少数原因之一。本研究旨在了解在动物模型中经胃肠外注射噬菌体后中和抗体是否会升高。本研究纳入了针对五种不同细菌的噬菌体，即 、 、 、 和 。这些噬菌体被分离、增殖和纯化。通过在各自的细菌菌苔上进行斑点试验来确认噬菌体的特异性。每周给五只兔子皮下注射纯化的噬菌体（10 PFU），持续六周。每周在下一次给药前采集血样。通过噬菌体中和试验检测抗体反应，随后使用双层琼脂覆盖法进行噬菌斑测定。通常在免疫后第3周后观察到抗噬菌体中和抗体升高。在免疫后3至5周可观察到噬菌体的完全中和。到第6周结束时，针对大多数注射的噬菌体，在1:1000和1:2000血清稀释度下可注意到噬菌体数量（PFU）进一步增加。观察到针对 特异性噬菌体的背景抗噬菌体中和抗体。然而，针对其他四种细菌特异性的噬菌体则不存在这种情况。噬菌体与哺乳动物宿主相互作用并诱导抗噬菌体中和抗体产生。然而，噬菌体的中和取决于重复给药和治疗持续时间。在第3周结束时可观察到中和抗体显著升高。因此，噬菌体可有效用于治疗持续时间少于2周的急性病例。然而，对于长期治疗，可能建议使用不同抗原性的噬菌体鸡尾酒。
