COMPARATIVE STUDY OF “KLIM-TEST” MONOCLONAL ANTIBODIES FOR PHENOTYPING OF PERIPHERAL BLOOD CELLS WITH MONOCLONALS FROM FOREIGN PRODUCERS

In view of wide-spread usage of flow cytometry (FC) in various areas of modern medicine, Russian market is still dominated by foreign manufacturers of monoclonal antibodies (MAbs). This problem
is accomplished by lack of information about Russian producers of MAbs. Objective: to compare a set of FC reagents produced by a Russian manufacturer with MAbs for FC from different foreign manufacturers. Venous blood served as biomaterial for this study. The parameters under study were measured by FC techniques. Comparative immunophenotyping was performed with MAbs from Becton Dickinson (USA), Lab Constant (Russia), and Beckman Coulter (USA). Virtually complete absence of systematic differences and a relatively small spread of values was revealed by means of Bland–Altman analysis. The values of parameters, as measured with different MAbs, did not differ from one another by significance level of 0.01, according to Friedman criterion. The following evidence has been yielded in present study: (1) data obtained with “Klim-Test” reagents do not exhidit any systematic differences, as compared with Mabs from foreign manufacturers, and
they show a relatively small scatter of results. “Klim-Test” reagents may be applied for Becton Dickinson and Beckman Coulter flow cytometers. (2) These reagents are equally effective at the levels of CD4-lymphocytes above 500 cells/mcL, and below these values. (3) Usage of “Klim-Test” reagents with a three-color label for CD45 cells (CD3-FITC/CD4-PE/CD45-PE-Sy5) corresponds to the standards for CD4-lymphocyte count in HIV-infected patients, thus allowing it to recommend this reagent for HIV-infection monitoring.

monoclonal antibodies, CD45 gating, double-platform approach

1. Бурместр Г.Р., Пецутто А. Наглядная иммунология: Пер. с англ. – М.: БИНОМ. Лаборатория знаний, 2007. – 320 с. Burmestr G.R. Naglyadnaya immunologiya [Visual immunology/ Trans. from angl: G.-R. Byrrmestr, A. Pettstso]. Moscow: BINOM. Laboratoriya znaniy – BINOM. Laboratory of Knowledge, 2007. 320 p.

2. ВИЧ-инфекция и СПИД. Клинические рекомендации / под редакцией В.В. Покровского. – 2-е изд., перераб. и доп. – М.: Геотар-Медиа, 2010. – 192 с. VICH-infektsiya i SPID. Klinicheskie rekomendatsii [Clinical recommendations under the editorship of V. Pokrovsky. - 2-e Izd.]. Moscow: Geotar-Media – Geotar-Media, 2010. 192 p.

3. ВИЧ-инфекция. Информационный бюллетень № 28 / под редакцией Н.Н.Ладной. – М., 2006. – 24 с. VICH-infektsiya. Informatsionnyy byulleten` №28/ pod redaktsiey N.N.Ladnoy [HIV-infection. Newsletter №28/ under the editorship of N.N.Ladnaya], Moscow, 2006. 24 p.

4. Гланц Стентон. Медико-биологическая статистика / Стентон Гланц. Пер. с англ. – М.: Практика, 1999. – 459 с. Glants Stenton. Mediko-biologicheskaya statistika [Trans. from angl.: Stanton Glantz. Medico-biological statistics]. Moscow: Praktika – Practice, 1999, 459 p.

5. Хоффман К., Рокшто Ю.К. Лечение ВИЧ-инфекции 2009. – М.: Р. Валент, 2012. – 736 с. Hoffman K. Lechenie VICH-infektsii 2009 [Treatment of HIV-2009/ Christian Hoffmann, Jrgen К.Rockstroh]. Moscow: R.Valent – R.Valent, 2012, 736 p.

6. Borkov G., Qibin L., Bentwich Z. Immune activation correlates better than HIV plasma viral load with CD4 T-cell decline during HIV infection // 8th European conference on clinical aspects and treatment of HIV-infection. Athens, 2001, abstr 535.

7. CDC Guideline for performing singl-platform CD4+T-cell determinations with CD45 gating forpersons infected with HIV. MMWR, Jan 31, 2003 / 52 (RRO2), pp. 1-13.

8. Eidukevicius R., Characiejus D., Janavicius R., Kazlauskaite N., Pasukoniene V., Mauricas M., Den Otter W. A method to estimate cell cycle time and growth fraction using bromodeoxyuridine-flow cytometry data from a single sample. BMC Cancer, 2005, vol. 5, p. 122.

9. Hazenberg M.D., Stuart J.W., Otto S.A. T-cell division in human immunodeficiency virus (HIV)-1 infection is mainly due to immune activation: a longitudinal analysis in patients before and during highly active antiretroviral therapy (HAART). Blood, 2000, vol. 95, pp. 249-255.

10. Rodrigues D.S, Salom o R., Kallas E.G. Profound peripheral T-lymphocyte depletion and activation in disseminated tuberculosis. Braz. J. Infect Dis., 2006, vol. 10, no. 1, pp. 59-61.

11. Schacker T.W., Bosch R.J., Bennett K., Pollard R., Robbins G.K, Collier A.C., Gulick R.M., Spritzler J., Mildvan D. Measurement of Na ve CD4 cells realiably predicts Potential for immune reconstitution in HIV. JAIDS, 2010, vol. 54, issue 1, pp. 59-62.

12. Schnizein-Bick C.T, Mandy F.F., O’Gorman M.R.G., Paxton H., Nicholson J.K., Hultin L.E., Gelman R.S., Wilkening C.L., Livnat D. Use of CD45 Gating in Three and Four-color Flow Cytometric Immunophenotyping: Guideline From The National Institute of Allergy and Infectious Diseases, Division of AIDS. Cytometry, 2002, vol. 50, no. 2, pp. 46-52.