Ukr.Biochem.J. 2019; Volume 91, Issue 1, Jan-Feb, pp. 30-37


Different mice inbred strains humoral immune response against human prostate-specific antigen

O. Yu. Galkin1,2, A. G. Komar3, O. B. Besarab1

1National Technical University of Ukraine “Igor Sikorsky Kyiv Polytechnic Institute”;
2Propharma Plant Ltd., Kyiv;
3Ukrainian Medical Center of Certification of Ministry of Health of Ukraine, Kyiv

Received: 21 August 2018; Accepted: 13 December 2018

The aim of the study was an investigation of humoral immune response to prostate-specific antigen (PSA) of different mice inbred strains for further development of recommendations for appropriate immunization schemes for monoclonal antibodies (McAbs) obtaining. The study was conducted using: Balb/c and NZB mice; PSA from human sperm (as immunogen). In case of booster immunization immunogen was previously diluted to the desired concentration (10 or 30 µg per 100 µl) in saline, and injected in the tail vein or intraperitoneally. In case of other immunizations, we prepared emulsion solution of immunogen with adjuvant to final concentration of 10 or 30 µg per 100 µl: PSA was dissolved in saline, the same volume of adjuvant was added, and mixture was thoroughly mixed to form a stable emulsion. When subcutaneous administration, total dose of 100 µl was divided into two equal parts and injected in the hind paw of mice. Intraperitoneal immunization was performed by single administration of 100 µl of emulsion. The level of specific antibodies was determined by titration of blood sera from animals in indirect ELISA. Series of experiments were conducted to determine the level of humoral response of mice of different inbred strains (Balb/c and NZB) for multi-stage immunization with different duration with different amounts of PSA (10 and 30 μg), with different immunoglobulin administration (intraperitoneal and subcutaneously), and with different adjuvants (Freund’s complete and incomplete adjuvants, FCA/FIA). Final booster immunization at a dose of 30 μg was performed either in the same manner as the previous administration of the immunogen, or intravenously in the physio­logical saline solution.  Dependencies of the humoral immune response of Balb/c and NZB mice against PSA on the route of administration of immunogen, the dose and duration of immunization were established. It was shown that intraperitoneal administration provided formation of higher titers of specific antibodies in case of both mice strains. Balb/c mice lines more rapidly responded to PSA, than an NZB mice (for all investigated schemes immunization). It was shown that the most effective immunization scheme was three times intraperitoneal administration with 10 µg of PSA for 8 weeks (the first immunization with FCA, and the rest – with FIA) and booster immunogen intravenous administration in saline solution.

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  1. Balk SP, Ko YJ, Bubley GJ. Biology of prostate-specific antigen. J Clin Oncol. 2003 Jan 15;21(2):383-91.  PubMed, CrossRef
  2. Lilja H. Biology of prostate-specific antigen. Urology. 2003 Nov;62(5 Suppl 1):27-33. PubMed, CrossRef
  3. Haythorn MR, Ablin RJ. Prostate-specific antigen testing across the spectrum of prostate cancer. Biomark Med. 2011 Aug;5(4):515-26. PubMed, CrossRef
  4. Ward AM, Catto JW, Hamdy FC. Prostate specific antigen: biology, biochemistry and available commercial assays. Ann Clin Biochem. 2001 Nov;38(Pt 6):633-51. PubMed, CrossRef
  5. Peehl DM. Prostate specific antigen role and function. Cancer. 1995;75(S7):2021-2026.
  6. Duskova K, Vesely S. Prostate Specific Antigen. Current clinical application and future prospects. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 2015; 159(1):018-026. CrossRef
  7. Loeb S, Catalona WJ. Prostate-specific antigen in clinical practice. Cancer Lett. 2007 Apr 28;249(1):30-9. PubMed, CrossRef
  8. Redchuk TA, Korotkevich NV, Kaberniuk AA, Oliinyk ES, Labyntsev AIu, Romaniuk SI, Kolibo DV, Busol VA, Komisarenko SV. Statistical analysis of distribution of antibody level against Mycobacterium bovis antigens for bovine tuberculosis diagnostics. Tsitol Genet. 2010 Sep-Oct;44(5):23-30. (In Russian). PubMed, CrossRef
  9. Romaniuk SI, Kolibo DV, Kavun EM, Radavskii IuL, Tkachenko TIu, Kramarev SA, Komissarenko SV. Specificity of antibodies to diphtheria toxin subunits in children with various forms of diphtheria infections. Ukr Biokhim Zhurn. 2001 Nov-Dec;73(6):73-6. (In Russian). PubMed
  10. Gavrilova ES, Bobrovnik SA, Sherriff G, Myslivets AA, Tarasov SA, Epstein OI. Novel approach to activity evaluation for release-active forms of anti-interferon-gamma antibodies based on enzyme-linked immunoassay. PLoS One. 2014 May 9;9(5):e97017.  PubMed, PubMedCentral, CrossRef
  11. Shtapenko OV, Hevkan II, Slyvchuk YuI, Syrvatka VY, Matvienko NM. Formation and properties of polymer nanolayers to enhance cell growth in vitro. Innov Biosyst Bioeng. 2018;2(2):105-109. CrossRef
  12. Galkin OYu, Savchenko AA, Nikitina KI, Dugan OM. Obtaining and study of properties of new monoclonal antibodies against human IgЕ. Ukr Biokhim Zhurn. 2013; 85(5): 81-87. (In Ukrainian).  PubMed, CrossRef
  13. Galkin OY, Besarab AB, Lutsenko TN. Characteristics of enzyme-linked immunosorbent assay for detection of IgG antibodies specific to Сhlamydia trachomatis heat shock protein (HSP-60). Ukr Biochem J. 2017 Jan-Feb;89(1):22-30. PubMed, CrossRef
  14. Galkin AYu, Kazmirchuk VE, Bondarenko LB. Different mice inbred strains humoral immune response to horseradish peroxidase. Proc Belarus State Univ. Physiol Biochem Mol Basis Funct Biosyst. 2014;9(1):51-55.
  15. Kozhemyakin YuM, Hromov OS, Filonenko MA, Sajfetdinova GA. Scientific and practical recommendations for the maintenance of laboratory animals and work with them. Kiev: Avicenna, 2002. 156 p. (In Ukrainian).
  16. Michel S, Deléage G, Charrier JP, Passagot J, Battail-Poirot N, Sibai G, Jolivet M, Jolivet-Reynaud C. Anti-free prostate-specific antigen monoclonal antibody epitopes defined by mimotopes and molecular modeling. Clin Chem. 1999 May;45(5):638-50. PubMed
  17. Nurmikko P, Väisänen V, Piironen T, Lindgren S, Lilja H, Pettersson K. Production and characterization of novel anti-prostate-specific antigen (PSA) monoclonal antibodies that do not detect internally cleaved Lys145-Lys146 inactive PSA. Clin Chem. 2000 Oct;46(10):1610-8. PubMed
  18. Leinonen J, Wu P, Stenman UH. Epitope mapping of antibodies against prostate-specific antigen with use of peptide libraries. Clin Chem. 2002 Dec;48(12):2208-16. PubMed
  19. Tijssen P. Practice and Theory of Enzyme Immunoassays. In: Laboratory Techniques in Biochemistry and Molecular Biology. 1985; 15: 674 p. CrossRef
  20. Galkin OYu, Gorshunov YuV, Besarab OB, Ivanova OM.  Development and characterization of highly informative ELISA for the detection of IgG and IgA antibodies to Сhlamydia trachomatis. Ukr Biochem J. 2018;90(3):70-83.  CrossRef
  21. Galkin OYu, Nikolaenko IV, Spivak MYa. Monoclonal antibodies to human immunoglobulins: features of obtaining and application. Immunol Allergol. 2005;1:3-9. (In Ukrainian).
  22. Frosch M, Görgen I, Boulnois GJ, Timmis KN, Bitter-Suermann D. NZB mouse system for production of monoclonal antibodies to weak bacterial antigens: isolation of an IgG antibody to the polysaccharide capsules of Escherichia coli K1 and group B meningococci. Proc Natl Acad Sci USA. 1985 Feb;82(4):1194-8. PubMed, PubMedCentral, CrossRef
  23. Vargas P, Cortés C, Vargas L, Rosemblatt M, Bono MR. Immunization with antigen-pulsed dendritic cells significantly improves the immune response to weak self-antigens. Immunobiology. 2006;211(1-2):29-36. PubMed, CrossRef
  24. Moe GR, Dave A, Granoff DM. Epitopes recognized by a nonautoreactive murine anti-N-propionyl meningococcal group B polysaccharide monoclonal antibody. Infect Immun. 2005 Apr;73(4):2123-8. PubMed, PubMedCentral, CrossRef
  25. Nussbaum G, Anandasabapathy S, Mukherjee J, Fan M, Casadevall A, Scharff MD. Molecular and idiotypic analyses of the antibody response to Cryptococcus neoformans glucuronoxylomannan-protein conjugate vaccine in autoimmune and nonautoimmune mice. Infect Immun. 1999 Sep;67(9):4469-76. PubMed, PubMedCentral
  26. Zhou H, Wang Y, Wang W, Jia J, Li Y, Wang Q, Wu Y, Tang J. Generation of monoclonal antibodies against highly conserved antigens. PLoS One. 2009 Jun 30;4(6):e6087. PubMed, PubMedCentral, CrossRef

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