Characteristics of antibody production to individual cytomegalovirus proteins in children with keratitis and uveitis

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Abstract

The clinical diagnosis of cytomegalovirus (CMV) eye pathology in children needs laboratory confirmation. The disadvantages of determining antibodies to CMV in serum in enzyme-linked immunosorbent assay (ELISA) include the presence of false positive and false negative results.

AIM: To determine the features of the synthesis of antibodies to the proteins of the tegument pp65, pp150, pp28 and the DNA-binding protein pp52 of cytomegalovirus, as well as to compare the diagnostic effectiveness of two laboratory methods of serodiagnostics of CMVI in children with uveitis and keratitis of different genesis: enzyme immunoassay and linear immunolysis.

MATERIAL AND METHODS: A total of 30 children (age 5–16 years) with uveitis (n=14) and keratitis (n=16) were included in the study. The IgM and IgG antibodies to the immediate early (IE) and late antigens of the virus (serological markers of primary, chronic, and reactivation of chronic infection) were determined in the ELISA. The IgG antibodies to individual recombinant CMV antigens containing only immunodominant protein fragments of viral antigens were studied in the line immunoassay (LIA): the main nonstructural IE protein, DNA-binding phosphoprotein p52, and phosphoproteins of the tegument p150, p65, and p28. The results of LIA were evaluated visually.

RESULTS: CMV infection (CMVI) in children with uveitis in ELISA was almost 2 times higher than that with keratitis (10/14%–71% vs. 6/16%–37.5%, p >0.05). Of the four positive results for detection of antibodies to IE antigen in ELISA, one was confirmed in LIA. In general, the discrepancy between the results of the determination of IgG antibodies to IE antigen in ELISA and LIA was noted in 13% of serum samples. LIA showed the increased frequency (p >0.05) and intensity (p <0.05) of antibody formation to viral antigens p65 and p52 in uveitis compared with keratitis, which confirmed the important role of CMVI in the pathogenesis of uveitis.

CONCLUSION: Both methods revealed a higher level of CMVI in children with uveitis than those with keratitis. IgG antibodies to IE-antigen serological markers of the reactivation of chronic CMVI have clinical importance because they serve as the basis for the appointment of antiviral therapy. Discrepancies between the results of ELISA and LIA in the detection of antibodies to IE antigen indicate the expediency of confirming the results of ELISA in LIA.

Thus, laboratory examination of sera in ELISA with subsequent analysis of antibodies to individual recombinant CMV antigens in LIA is an effective, highly sensitive, and highly specific method to verify CMVI and determine its activity. The most informative action is the determination of IgG antibodies to recombinant CMV antigens: IE antigens, p65, and p52.

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About the authors

Galina I. Krichevskaya

Helmholtz National Medical Research Center of Eye Diseases

Author for correspondence.
Email: gkri@yandex.ru
ORCID iD: 0000-0001-7052-3294
SPIN-code: 6808-0922

MD, PhD

Russian Federation, 14/19, Sadovaya-Chernogriazskaya street, Moscow, 105062

Lyudmila A. Katargina

Helmholtz National Medical Research Center of Eye Diseases

Email: katargina@igb.ru
ORCID iD: 0000-0002-4857-0374
Scopus Author ID: 137428

MD, Dr. of Sci. (Med.), professor

Russian Federation, 14/19, Sadovaya-Chernogriazskaya street, Moscow, 105062

Galina I. Alatortseva

I.I. Mechnikov Research Institute for Vaccines and Sera

Email: gkri@yandex.ru
ORCID iD: 0000-0001-9887-4061
SPIN-code: 8911-3000

PhD (Biology)

Russian Federation, Moscow

Vera V. Dotsenko

I.I. Mechnikov Research Institute for Vaccines and Sera

Email: gkri@yandex.ru
ORCID iD: 0000-0002-5866-944X
SPIN-code: 9250-0131

PhD (Biology), Senior Researcher

Russian Federation, Moscow

Lyubov N. Nesterenko

I.I. Mechnikov Research Institute for Vaccines and Sera

Email: gkri@yandex.ru
ORCID iD: 0000-0002-3825-3906
SPIN-code: 6819-7980

PhD (Chemistry)

Russian Federation, Moscow

References

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  2. Kalejta RF. Tegument proteins of human cytomegalovirus. Microbiol Mol Biol Rev. 2008;72(2):249-265, table of contents. doi: 10.1128/MMBR.00040-07
  3. Biolatti M, Dell’Oste V, De Andrea M, Landolfo S. The human cytomegalovirus tegument protein pp65 (pUL83): a key player in innate immune evasion. New Microbiol. 2018;41(2):87–94.
  4. Beqaj SH, Lerner AM, Fitzgerald JT. Immunoassay with cytomegalovirus early antigens from gene products p52 and CM2 (UL44 and UL57) detects active infection in patients with chronic fatigue syndrome. J Clin Pathol. 2008;61(5):623–626. doi: 10.1136/jcp.2007.050633
  5. Han MS, Choi EH, Lee HJ, et al. Cytomegalovirus disease in a retinoblastoma cohort: The role of preemptive screening. Pediatr Blood Cancer. 2020;67(3):e28101. doi: 10.1002/pbc.28101
  6. Galeeva GZ, Rascheskov AY. Cytomegalovirus keratitis in child (case study). Practical medicine. 2016;(2–1):128–130. (In Russ).
  7. Neroev VV, Krichevskaya GI, Alatortseva GI, et al. Features of IgG-antibodies production to individual Cytomegalovirus proteins in various eye diseases (age-related macular degeneration and central serous chorioretinopathy). Russian Journal of Infection and Immunity. 2021;11(3):530-–538. (In Russ). doi: 10.15789/2220-7619-foi-1424

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Свидетельство о регистрации СМИ ПИ № ФС 77 - 28266 от 17.05.2007 г. выдано Федеральной службой по надзору в сфере связи, информационных технологий и массовых коммуникаций (Роскомнадзор). 
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