噬菌体治疗泛耐药肺炎克雷伯菌肺部感染的临床应用及效果初探

doi:10.3969/j.issn.1674-8115.2021.09.022

上海交通大学学报(医学版) ›› 2021, Vol. 41 ›› Issue (9): 1272-1276.doi: 10.3969/j.issn.1674-8115.2021.09.022

• 短篇论著 • 上一篇

噬菌体治疗泛耐药肺炎克雷伯菌肺部感染的临床应用及效果初探

李莉莎¹^,²(), 李建辉², 何斌², 吴楠楠², 朱同玉², 郭晓奎²^,³, 陈峥宏¹()

^1.贵州医科大学基础医学院微生物学教研室，贵阳 550025
^2.上海市公共卫生临床中心，上海噬菌体研究所，上海 201508
^3.上海交通大学医学院国家热带病研究中心全球健康学院，上海 200025

收稿日期:2021-03-10 出版日期:2021-08-24 发布日期:2021-08-24
通讯作者: 陈峥宏 E-mail:xiaxiaday@hotmail.com;chenzhenghong@gmc.edu.cn
作者简介:李莉莎（1993—），女，硕士生；电子信箱：xiaxiaday@hotmail.com。
基金资助:
上海市自然科学基金(17ZR1415900)

Clinical application and effect of phage on the treatment of pulmonary infection by pan-drug resistant Klebsiella pneumoniae

Li-sha LI¹^,²(), Jian-hui LI², Bin HE², Nan-nan WU², Tong-yu ZHU², Xiao-kui GUO²^,³, Zheng-hong CHEN¹()

^1.Department of Microbiology, School of Basic Medical Science, Guizhou Medical University, Guiyang 550025, China
^2.Shanghai Institute of Phage, Shanghai Public Health Clinical Center, Shanghai 201508, China
^3.School of Global Health of Chinese Center for Tropical Diseases Research, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China

Received:2021-03-10 Online:2021-08-24 Published:2021-08-24
Contact: Zheng-hong CHEN E-mail:xiaxiaday@hotmail.com;chenzhenghong@gmc.edu.cn
Supported by:
Natural Science Foundation of Shanghai(17ZR1415900)

摘要/Abstract

摘要：

目的·探讨使用噬菌体雾化吸入治疗泛耐药肺炎克雷伯菌肺部感染的用药频率及效果。方法·在治疗开始前1 h和使用噬菌体雾化吸入治疗肺部感染过程中每小时，取患者痰液样本进行细菌分离纯化及质谱鉴定，比较细菌裂解谱的异同；同时，于治疗开始前1 h，第一次治疗开始后2、4 h取血液样本进行细菌培养及噬菌体量的测定。结果·噬菌体经雾化吸入后2 h在体内持续增殖，3 h后噬菌体量下降，4 h后进行第二次治疗，噬菌体在体内的增殖规律与第一次相同；第三次使用噬菌体治疗时，噬菌体仅有少量增殖。噬菌体治疗后患者痰液中仍分离到肺炎克雷伯菌，但分离株的噬菌体裂解谱发生了变化，患者临床症状改善。结论·噬菌体能够有效针对靶细菌进行杀灭并改善患者的临床症状。泛耐药肺炎克雷伯菌肺部感染患者经2次雾化吸入治疗即可达到噬菌体使用的最佳效果。噬菌体治疗后病原菌菌株发生了变化。

关键词: 噬菌体, 泛耐药, 肺炎克雷伯菌, 治疗应用

Abstract:

Objective·To explore the therapeutic frequency and effect of phage treatment on pan-drug resistant Klebsiella pneumoniae by aerosolized phage.

Methods·The patient's sputum was taken 1 h before the start of treatment and every hour during the treatment of pulmonary infection using nebulized inhaled phage. The sputum samples were subjected to bacterial isolation, purification and mass spectrometry identification. The lytic spectrum of different bacterial isolations was compared. And blood samples for bacteria were taken 1 h before the start of the treatment and 2 h and 4 h after the first treatment for bacterial culture and phage volume determination.

Results·The phage continued to proliferate in vivo for 2 h, and decreased after 3 h. When the second phage treatment was performed after 4 h, the pattern of phage proliferation in vivo was the same as the first time. During the third treatment, the phage was proliferated slightly in vivo. After phage treatment, Klebsiella pneumoniae was still isolated in the patient's sputum. However, the phage cleavage spectrum of the isolations was different, and the patient's clinical symptoms improved.

Conclusion·Phages can effectively target infectious bacteria and kill them. The clinical symptoms of patient improve. Twice aerosolized phage treatment for the patient with pulmonary infection is enough to achieve the best effect. The isolated bacterial strain is different post phage treatment.

Key words: phage, pan-drug resistant, Klebsiella pneumoniae, therapeutic application

中图分类号:

Q939.48

李莉莎, 李建辉, 何斌, 吴楠楠, 朱同玉, 郭晓奎, 陈峥宏. 噬菌体治疗泛耐药肺炎克雷伯菌肺部感染的临床应用及效果初探[J]. 上海交通大学学报(医学版), 2021, 41(9): 1272-1276.

Li-sha LI, Jian-hui LI, Bin HE, Nan-nan WU, Tong-yu ZHU, Xiao-kui GUO, Zheng-hong CHEN. Clinical application and effect of phage on the treatment of pulmonary infection by pan-drug resistant Klebsiella pneumoniae[J]. JOURNAL OF SHANGHAI JIAOTONG UNIVERSITY (MEDICAL SCIENCE), 2021, 41(9): 1272-1276.

图/表 6

图1 患者使用抗生素和噬菌体的治疗经过Note：MEM—meropenem; AMK—amikacin; SCF—cefoperazone sodium and sulbactam sodium; TGC—tigecycline; MDE—metronidazole; PHM—fosfomycin; MXF—moxifloxacin; ZAVICEFTA—ceftazidime and avitbatan sodium; MF—mikafen sodium; AMB—amphotericin B.

Fig 1 Course of antibiotics and phage treatment for the patient

表1 痰液与血液样本采集方案

Tab 1 Scheme for collection of sputum and blood samples

Item	Time/h	Sputum sample	Blood sample
Before treatment	0	#1	#1
The first treatment	1	#2	/
	2	#3	#2
	3	#4	/
	4	#5	#3
The second treatment	5	#6	/
	6	#7	/
	7	#8	/
	8	#9	/
The third treatment	9	#10	/
	10	#11	/
	11	#12	/
	12	#13	/
The fourth treatment	13	#14	/
Four hours after the 4^th treatment	17	#15	#4

图2 临床分离株KpLJW-1的噬菌体裂解谱

Fig 2 Phage lysis spectrum of clinical isolate KpLJW-1

图3 痰液中噬菌体定量及细菌生长情况

Fig 3 Titer of phages and the survival of bacteria in sputum

图4 临床分离株KpLJW-2（A）和KpLJW-3（B）的噬菌体裂解谱

Fig 4 Phage lysis spectrum of clinical isolates KpLJW-2（A）and KpLJW-3（B）

图5 噬菌体治疗前（A)、后(B)患者肺部CT平扫对比Note：The yellow arrows indicate areas of inflammatory exudation and pleural effusion.

Fig 5 Lung CT scan of the patient before (A) and after (B) phage treatment

参考文献 12

1	Cai RP, Wu M, Zhang H, et al. A smooth-type, phage-resistant Klebsiella pneumoniae mutant strain reveals that OmpC is indispensable for infection by phage GH-K3[J]. Appl Environ Microbiol, 2018, 84(21): e01585-e01518.
2	Tomás JM, Benedí VJ, Jofre JT. Identification of the cell surface receptor for FC₃-2, FC₃-3 and FC₃-6 bacteriophages from Klebsiella pneumoniae[J]. FEMS Microbiol Lett, 1987, 41(3): 223-228.
3	Goldfarb T, Sberro H, Weinstock E, et al. BREX is a novel phage resistance system widespread in microbial genomes[J]. EMBO J, 2015, 34(2): 169-183.
4	Leung CYJ, Weitz JS. Modeling the synergistic elimination of bacteria by phage and the innate immune system[J]. J Theor Biol, 2017, 429: 241-252.
5	Malik DJ, Sokolov IJ, Vinner GK, et al. Formulation, stabilisation and encapsulation of bacteriophage for phage therapy[J]. Adv Colloid Interface Sci, 2017, 249: 100-133.
6	Bakhrushina EO, Anurova MN, Aleshkin AV, et al. Development of the composition and pharmacokinetic studies of suppositories with combined substance of bacteriophages[J]. J Drug Deliv Sci Technol, 2020, 59: 101841.
7	Dąbrowska K. Interaction of bacteriophages with the immune system: induction of bacteriophage-specific antibodies[J]. Methods Mol Biol, 2018, 1693: 139-150.
8	Sinha A, Maurice CF. Bacteriophages: uncharacterized and dynamic regulators of the immune system[J]. Mediat Inflamm, 2019, 2019: 3730519.
9	Azam AH, Tanji Y. Bacteriophage-host arm race: an update on the mechanism of phage resistance in bacteria and revenge of the phage with the perspective for phage therapy[J]. Appl Microbiol Biotechnol, 2019, 103(5): 2121-2131.
10	Wright RCT, Friman VP, Smith MCM, et al. Resistance evolution against phage combinations depends on the timing and order of exposure[J]. mBio, 2019, 10(5): e01652-e01619.
11	Oechslin F. Resistance development to bacteriophages occurring during bacteriophage therapy[J]. Viruses, 2018, 10(7): E351.
12	Kumaran D, Taha M, Yi QL, et al. Does treatment order matter? investigating the ability of bacteriophage to augment antibiotic activity against Staphylococcus aureus biofilms[J]. Front Microbiol, 2018, 9: 127.

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噬菌体治疗泛耐药肺炎克雷伯菌肺部感染的临床应用及效果初探

Clinical application and effect of phage on the treatment of pulmonary infection by pan-drug resistant Klebsiella pneumoniae

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