Identification and analysis of host proteins in the cyst wall and cyst fluid of Echinococcus granulosus in human infection

doi:10.12140/j.issn.1000-7423.2023.06.003

Abstract

Abstract:

Objective To further elucidate the interaction between Echinococcus granulosus and the host by identifying the host protein components in the cyst wall and cyst fluid of E. granulosus. Methods The active and intact echinococcal cysts removed surgically from 10 patients with E. granulosus were collected from the Department of Hepatobiliary Surgery, Ningxia Medical University General Hospital in 2022. Blood samples were also taken from the same patients. The transparent cyst fluid in the ascus was extracted aseptically, and centrifuged to collect the supernatant; the inner wall of the ascus was repeatedly washed, cut into pieces, lysed and homogenised to collect the supernatant. The bicinchoninic acid (BCA) protein quantification kit was used to extract the cyst fluid and cyst wall total protein. The cyst wall total protein lane was cut out after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) electrophoresis. The significant protein bands were analysed by mass spectrometry and Western blotting analysis. Quantitative analysis was performed to determine the biochemical component concentrations in the patient serum and cyst fluid. Results The liver function of 10 patients with echinococcosis granulosus was Child-Pugh A or B. The diameter of the lesion ranged from 2.9 to 16.3 cm, which were classified as CE2 (multiple subcysts) or CE3 (collapsed internal capsule) according to the WHO-IWGE classification. SDS-PAGE electrophoresis analysis results suggested that the echinococcal cyst wall protein showed clear protein bands at the relative molecular mass (M_r) of 70 000, 50 000, and 25 000, and the cyst fluid protein showed clear bands at M_r 70 000. Protein bands. The protein spectrum and Western blotting results showed that the protein with a relative molecular mass of approximately M_r 70 000 contained human albumin, the target protein with a relative molecular mass of approximately M_r 50 000 was a human IgG heavy chain, and the target protein with a relative molecular mass of approximately M_r 25 000 was a human IgG light chain. Among the various biochemical indicators, the concentrations of K⁺ in cyst fluid and serum were (5.53 ± 0.86) mmol/L and (3.92 ± 0.33) mmol/L respectively. The albumin concentrations were (0.57 ± 0.46) g/L and (39.42 ± 2.77) g/L. The cholesterol concentrations were (0.03 ± 0.02) mmol/L and (3.79 ± 1.53) mmol/L respectively. The concentration differences between the three in cyst fluid and serum were statistically significant (t = 5.56, 43.71, 7.74; P < 0.05). Conclusion Host-derived serum albumin and antibodies are found of comparatively higher amont in the cyst wall and cyst fluid of E. granulosus.

Key words: Echinococcus granulosus, Albumin, Antibodies, Cyst wall, Cyst fluid

CLC Number:

R532.32

LI Tao, LI Zihua, ZHANG Cuiying, ZHAO Wei. Identification and analysis of host proteins in the cyst wall and cyst fluid of Echinococcus granulosus in human infection[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2023, 41(6): 677-682.

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项目 Project	囊液 Cyst fluid	血清 Serum	t值 t values	P值 P values
渗透压/mOsm Osmotic pressure/mOsm	313.50 ± 73.33	301.59 ± 6.20	0.52	> 0.05
Na⁺浓度/mmol·L^-1Na⁺ concentration/mmol·L^-1	143.40 ± 25.61	141.43 ± 2.42	0.24	> 0.05
K⁺浓度/mmol·L^-1K⁺ concentration/mmol·L^-1	5.53 ± 0.86	3.92 ± 0.33	5.56	< 0.05
Cl^-浓度/mmol·L^-1Cl^- concentration/mmol·L^-1	101.56 ± 12.42	105.35 ± 3.75	0.92	> 0.05
葡萄糖浓度/mmol·L^-1 Glucose concentration/mmol·L^-1	3.07 ± 2.61	4.65 ± 0.26	1.91	> 0.05
白蛋白浓度/g·L^-1 Albumin concentration/g·L^-1	0.57 ± 0.46	39.42 ± 2.77	43.71	< 0.05
胆固醇浓度/mmol·L^-1 Cholesterol concentration/mmol·L^-1	0.03 ± 0.02	3.79 ± 1.53	7.74	< 0.05