BLOOD LEUKOCYTES PHENOTYPING BY HEMATOFLOW METHOD IN PATIENTS WITH ONYCHOMYCOSIS

Cover Page


Cite item

Full Text

Abstract

The aim of the investigation was to evaluate the information content of Hematoflow method in the pathogenetic significance in determining the violations of the cellular responses of innate and adaptive immunity, as well as the decision on the appointment of immunotropic treatment of patients with onychomycosis. The study involved 42 patients with onychomycosis feet and hands/feet at the age of 20–45 years before the appointment of a systemic antifungal therapy. The diagnosis of mycosis, onychomycosis was confirmed by microscopic examination of the fragments of the damaged nail plate. The growth of the fungus culture on special media was observed in 64% of patients. 24 healthy persons were examined as controls. A study of the phenotype of white blood cells was performed on a dual-platform technology hematology analyzer and flow cytometry using a set of antibodies Cytodiff: CD36-FITC, CD2-PE, CD294(CRTH2)-PE, CD19-ECD, CD16-PC5 и CD45-PC7. The phenotypic composition evaluation of the white blood cells by the Hematoflow method allowed to establish in patients with onychomycosis the violation cellular innate and adaptive immunity. Minor changes were detected in the population composition of granule cells in the peripheral blood of patients manifested to an increase in the content of the young and segmented granulocytes. When monotsitopeniya patients with onychomycosis increases the content of the «classic» monocytes and decreases the level of «non-classical» monocytes. Changes in the composition of blood monocytes subpopulation identified in patients with the infection lasting up to 3 years and stored in the course of the disease. The most pronounced changes were found in patients with onychomycosis by the performance of adaptive immunity. Lymphopenia in these patients is realized by reducing the number of immature and mature B-cell, but by increasing the content of T-lymphocytes. Moreover, if the content of immature B-cells have decreased in patients with a duration of infection of up to 3 years, the change in the number of mature Tand B-lymphocytes detected during disease duration 3–10 and 10 years. These changes in the content of Tand B-lymphocytes reflect immunopathogenetic processes and determine the importance of Tand B-cell immunity in onychomycosis. In general, the Hematoflow method is informative in assessing violations the cell of innate and adaptive immunity. It allows to evaluate the severity of the immunopathological process mechanism and the level of damage to the immune system, can recommend its use for a personalized approach to the appointment immunotropic treatment. 

About the authors

A. A. Savchenko

Research Institute of Medical Problems of the North, Krasnoyarsk, Russian Federation

Author for correspondence.
Email: fake@neicon.ru

PhD, MD (Medicine), Professor, Head of the Laboratory of Molecular-Cell Physiology and Pathology, Research Institute of Medical Problems of the North, Krasnoyarsk, Russian Federation

Russian Federation

A. G. Borisov

Research Institute of Medical Problems of the North, Krasnoyarsk, Russian Federation

Email: fake@neicon.ru

PhD (Medicine), Leading Researcher, Laboratory of Molecular-Cell Physiology and Pathology, Research Institute of Medical Problems of the North, Krasnoyarsk, Russian Federation

Russian Federation

E. N. Anisimova

Krasnoyarsk State Medical University named after prof. V.F. Voino-Yasenetsky, Krasnoyarsk, Russian Federation

Email: fake@neicon.ru

PhD (Medicine), Head of the Department, Krasnoyarsk State Medical University named after prof. V.F. Voino-Yasenetsky, Krasnoyarsk, Russian Federation

Russian Federation

V. D. Belenyuk

Research Institute of Medical Problems of the North, Krasnoyarsk, Russian Federation

Email: fake@neicon.ru

PhD Candidate, Research Institute of Medical Problems of the North, Krasnoyarsk, Russian Federation

Russian Federation

I. V. Kudryavtsev

Institute of Experimental Medicine, St. Petersburg, Russian Federation
Far Eastern Federal University, Vladivostok, Russian Federation

Email: fake@neicon.ru

PhD (Biology), Senior Researcher, Laboratory of Immunology, Research Institute of Experimental Medicine, St. Petersburg; Senior Researcher, Department of Fundamental Medicine, Far Eastern Federal University, Vladivostok, Russian Federation

Russian Federation

I. V. Reshetnikov

South Ural State Medical University, Cheljabinsk, Russian Federation

Email: fake@neicon.ru

Biologist of the Immunology Laboratory, South Ural State Medical University, Cheljabinsk, Russian Federation

Russian Federation

S. V. Kvjatkovskaja

South Ural State Medical University, Cheljabinsk, Russian Federation

Email: fake@neicon.ru

PhD (Medicine), Head of the Laboratory, South Ural State Medical University, Cheljabinsk, Russian Federation

Russian Federation

V. Je. Cejlikman

South Ural State Medical University, Cheljabinsk, Russian Federation

Email: fake@neicon.ru

PhD, MD (Biology), Professor, Head of the Department South Ural State Medical University, Cheljabinsk, Russian Federation

Russian Federation

A. N. Zorin

Krasnoyarsk Regional sexually transmitted infection clinic No. 1, Krasnoyarsk, Russian Federation

Email: fake@neicon.ru

Clinical Mycologist, Krasnoyarsk Regional Sexually Transmitted Infection Clinic No. 1, Krasnoyarsk, Russian Federation

Russian Federation

References

  1. Борисов А.Г. Клиническая характеристика нарушения функции иммунной системы // Медицинская иммунология. 2013. Т. 15, № 1. С. 45–50. [Borisov A.G. Clinical characteristics of the dysfunction of the immune system. Meditsinskaya immunologiya = Medical Immunology (Russia), 2013, vol. 15, no. 1, pp. 45–50. doi: 10.15789/1563-0625-2013-1-45-50 (In Russ.)]
  2. Борисов А.Г., Савченко А.А., Смирнова С.В. К вопросу о классификации нарушений функционального состояния иммунной системы // Сибирский медицинский журнал. 2008. Т. 23, № 3. С.13–18. [Borisov A.G., Savchenko A.A., Smirnova S.V. On the classification of violations of the functional state of the immune system. Sibirskii meditsinskii zhurnal = Siberian Medical Journal, vol. 23, no. 3, pp. 13–18. (In Russ.)]
  3. Васенова В.Ю., Пичугин А.В., Бутов Ю.С, Атауллаханов Р.И. Влияние комплексной терапии онихомикоза на клинико-иммунологические параметры. Cообщение 3 // Российский журнал кожных и венерических болезней. 2008. № 2. С. 48–51. [Vasenova V.Ju., Pichugin A.V., Butov Ju.S., Ataullahanov R.I. Influence of complex therapy for onychomycosis clinical and immunological parameters. Message 3. Rossiiskii zhurnal kozhnykh i venericheskikh boleznei = Russian Journal of Skin and Venereal Diseases, 2008, no. 2, pp. 48–51. (In Russ.)]
  4. Васенова В.Ю., Атауллаханов Р.И., Пичугин А.В., Бутов Ю.С. Особенности иммунного статуса больных онихомикозом. Сообщение 1 // Российский журнал кожных и венерических болезней. 2007. № 4. С. 63–66. [Vasenova V.Ju., Ataullahanov R.I., Pichugin A.V., Butov Ju.S. Features of the immune status of patients with onychomycosis. Message 1. Rossiiskii zhurnal kozhnykh i venericheskikh boleznei = Russian Journal of Skin and Venereal Diseases, 2007, no. 4, pp. 63–66. (In Russ.)]
  5. Головкин А.С., Матвеева В.Г., Кудрявцев И.В., Григорьев Е.В., Великанова Е.А., Байракова Ю.В. Субпопуляции моноцитов крови при неосложненном течении периоперационного периода коронарного шунтирования // Медицинская иммунология. 2012. Т. 14, № 4–5. С. 305–312. [Golovkin A.S., Matveeva V.G., Kudryavtsev I.V., Grigor’ev E.V., Velikanova E.A., Bajrakova Ju.V. Blood monocyte subpopulations during uncomplicated coronary artery bypass surgery. Meditsinskaya immunologiya = Medical Immunology (Russia), 2012, vol. 14, no. 4–5, pp. 305–312. doi: 10.15789/1563-0625-2012-4-5-305-312 (In Russ.)]
  6. Зурочка А.В., Хайдуков С.В., Кудрявцев И.В., Черешнев В.А. Проточная цитометрия в медицине и биологии. Екатеринбург: Редакционно-издательский отдел Уральского отделения РАН, 2013. 552 с. [Zurochka A.V., Hajdukov S.V., Kudryavtsev I.V., Chereshnev V.A. Protochnaya tsitometriya v meditsine i biologii [Flow cytometry in medicine and biology]. Ekaterinburg: Publishing department of the Ural Branch of the Russian Academy of Sciences, 2013. 552 p.]
  7. Климко Н.Н., Козлова Я.И., Хостелиди С.Н., Шадривова О.В., Борзова Ю.В., Васильева Н.В. Распространенность тяжелых и хронических микотических заболеваний в Российской Федерации по модели Life program // Проблемы медицинской микологии. 2014. Т. 16, № 1. С. 3–8. [Klimko N.N., Kozlova Ja.I., Hostelidi S.N., Shadrivova O.V., Borzova Ju.V., Vasil’eva N.V. The incidence of severe and chronic mycotic diseases in the Russian Federation on the model of Life program. Problemy meditsinskoi mikologii = Problems of Medical Mycology, 2014, vol. 16, no. 1, pp. 3–8. (In Russ.)]
  8. Савченко А.А., Борисов А.Г., Модестов А.А., Мошев А.В., Кудрявцев И.В., Тоначева О.Г., Кощеев В.Н. Фенотипический состав и хемилюминесцентная активность моноцитов у больных почечноклеточным раком // Медицинская иммунология. 2015. Т. 17, № 2. С. 141–150. [Savchenko A.A., Borisov A.G., Modestov A.A., Moshev A.V., Kudryavtsev I.V., Tonacheva O.G., Koshheev V.N. The phenotypic composition and chemiluminescent activity of monocytes in patients with renal cell cancer. Meditsinskaya immunologiya = Medical Immunology (Russia), 2015, vol. 17, no. 2, pp. 141–150. doi: 10.15789/1563-0625-2015-2-141-150 (In Russ.)]
  9. Савченко А.А., Модестов А.А., Мошев А.В., Тоначева О.Г., Борисов А.Г. Цитометрический анализ NKи NKT-клеток у больных почечноклеточным раком // Российский иммунологический журнал. 2014. Т. 8 (17), № 4. C. 1012–1018. [Savchenko A.A., Modestov A.A., Moshev A.V., Tonacheva O.G., Borisov A.G. The cytometric analysis NKand NKT-cells in patients with renal cell cancer. Rossiiskii immunologicheskii zhurnal = Russian Journal of Immunology, 2014, vol. 8 (17), no. 4, pp. 1012–1018. (In Russ.)]
  10. Сергеев Ю.В., Касихина Е.И. Онихомикозы: современные подходы к лечению // Вестник дерматологии и венерологии. 2009. № 5. С. 117–119. [Sergeev Ju.V., Kasikhina E.I. Onychomycosis: modern approaches to treatment. Vestnik dermatologii i vene rologii = Journal of Dermatology and Venereology, 2009, no. 5, pp. 117–119. (In Russ.)]
  11. Ameen M., Lear J.T., Madan V., Mohd Mustapa M.F., Richardson M. British Association of Dermatologists’ guidelines for the management of onychomycosis 2014. Br. J. Dermatol., 2014, vol. 171, no. 5, pp. 937–958. doi: 10.1111/bjd.13358
  12. Appleby L.J., Nausch N., Midzi N., Mduluza T., Allen J.E., Mutapi F. Sources of heterogeneity in human monocyte subsets. Immunol. Lett., 2013, vol. 152, no. 1, pp. 32–41. doi: 10.1016/j.imlet.2013.03.004
  13. Baraldi A., Jones S.A., Guesné S., Traynor M.J., McAuley W.J., Brown M.B., Murdan S. Human nail plate modifications induced by onychomycosis: implications for topical therapy. Pharm. Res., 2015, vol. 32, no. 5, pp. 1626–1633. doi: 10.1007/s11095-014-1562-5
  14. Bhatnagar N., Ahmad F., Hong H.S., Eberhard J., Lu I.N., Ballmaier M., Schmidt R.E., Jacobs R., Meyer-Olson D. FcγRIII (CD16)-mediated ADCC by NK cells is regulated by monocytes and FcγRII (CD32). Eur. J. Immunol., 2014, vol. 44, no. 11, pp. 3368–3379. doi: 10.1002/eji.201444515
  15. Brasch J., Köppl G. Persisting onychomycosis caused by Fusarium solani in an immunocompetent patient. Mycoses, 2009, vol. 52, no. 3, pp. 285–286. doi: 10.1111/j.1439-0507.2008.01591.x.
  16. Bruserud O. Bidirectional crosstalk between platelets and monocytes initiated by Toll-like receptor: an important step in the early defense against fungal infections? Platelets, 2013, vol. 24, no. 2, pp. 85–97. doi: 10.3109/09537104.2012.678426
  17. Bunyaratavej S., Pattanaprichakul P., Leeyaphan C., Chayangsu O., Bunyaratavej S., Kulthanan K. Onychomycosis: a study of selfrecognition by patients and quality of life. Indian J. Dermatol. Venereol. Leprol., 2015, vol. 81, no. 3, pp. 270–274. doi: 10.4103/0378-6323.154796
  18. Burbano C., Vasquez G., Rojas M. Modulatory effects of CD14+CD16++ monocytes on CD14++CD16– monocytes: a possible explanation of monocyte alterations in systemic lupus erythematosus. Arthritis Rheumatol., 2014, vol. 66, no. 12, pp. 3371–3381. doi: 10.1002/art.38860
  19. Döbel T., Kunze A., Babatz J., Tränkner K., Ludwig A., Schmitz M., Enk A., Schäkel K. FcγRIII (CD16) equips immature 6-sulfo LacNAc-expressing dendritic cells (slanDCs) with a unique capacity to handle IgG-complexed antigens. Blood, 2013, vol. 121, no. 18, pp. 3609–3618. doi: 10.1182/blood-2012-08-447045
  20. Hristov M., Schmitz S., Nauwelaers F., Weber C. A flow cytometric protocol for enumeration of endothelial progenitor cells and monocyte subsets in human blood. J. Immunol. Methods. 2012, vol. 381, no. 1–2, pp. 9–13. doi: 10.1016/j.jim.2012.04.003
  21. Jo Y., Kim S.H., Koh K., Park J., Shim Y.B., Lim J., Kim Y., Park Y.J., Han K. Reliable, accurate determination of the leukocyte differential of leukopenic samples by using Hematoflow method. Korean J. Lab. Med., 2011, vol. 31, no. 3, pp. 131–137. doi: 10.3343/kjlm.2011.31.3.131
  22. Kahng J., Kim Y., Kim M., Oh E.J., Park Y.J., Han K. Flow cytometric white blood cell differential using CytoDiff is excellent for counting blasts. Ann. Lab. Med., 2015, vol. 35, no. 1, pp. 28–34. doi: 10.3343/alm.2015.35.1.28
  23. Kaya T.I., Eskandari G., Guvenc U., Gunes G., Tursen U., Burak Cimen M.Y., Ikizoglu G. CD4+CD25+ Treg cells in patients with toenail onychomycosis. Arch. Dermatol. Res., 2009, vol. 301, no. 10, pp. 725–729. doi: 10.1007/s00403-009-0941-y
  24. Kim A.H., Lee W., Kim M., Kim Y., Han K. White blood cell differential counts in severely leukopenic samples: a comparative analysis of different solutions available in modern laboratory hematology. Blood Res., 2014, vol. 49, no. 2, pp. 120–126. doi: 10.5045/br.2014.49.2.120
  25. Leelavathi M., Noorlaily M. Onychomycosis nailed. Malays Fam. Physician., 2014, vol. 9, no. 1, pp. 2–7.
  26. Lipner S.R., Scher R.K. Onychomycosis: current and investigational therapies. Cutis, 2014, vol. 94, no. 6, pp. 21–24.
  27. Rosen T., Friedlander S.F., Kircik L., Zirwas M.J., Stein Gold L., Bhatia N., Gupta A.K. Onychomycosis: epidemiology, diagnosis, and treatment in a changing landscape. J. Drugs Dermatol., 2015, vol. 14, no. 3, pp. 223–233.
  28. Skrzeczyńska-Moncznik J., Bzowska M., Loseke S., Grage-Griebenow E., Zembala M., Pryjma J. Peripheral blood CD14high CD16+ monocytes are main producers of IL-10. Scand. J. Immunol., 2008, vol. 67, no. 2, pp. 152–159. doi: 10.1111/j.1365-3083.2007.02051.x
  29. Trzeciak-Ryczek A., Tokarz-Deptuła B., Deptuła W. Antifungal immunity in selected fungal infections. Postepy Hig. Med. Dosw., 2015, vol. 69, pp. 469–474 doi: 10.5604/17322693.1148747
  30. Wüthrich M., Brandhorst T.T., Sullivan T.D., Filutowicz H., Sterkel A., Stewart D., Li M., Lerksuthirat T., LeBert V., Shen Z.T., Ostroff G., Deepe G.S. Jr, Hung C.Y., Cole G., Walter J.A., Jenkins M.K., Klein B. Calnexin induces expansion of antigenspecific CD4(+) T cells that confer immunity to fungal ascomycetes via conserved epitopes. Cell Host Microbe, 2015, vol. 17, no. 4, pp. 452–465. doi: 10.1016/j.chom.2015.02.009
  31. Ziegler-Heitbrock L., Ancuta P., Crowe S., Dalod M., Grau V., Hart D.N., Leenen P.J., Liu Y.J., MacPherson G., Randolph G.J., Scherberich J., Schmitz J., Shortman K., Sozzani S., Strobl H., Zembala M., Austyn J.M., Lutz M.B. Nomenclature of monocytes and dendritic cells in blood. Blood, 2010, vol. 116, no. 16, pp. 74–80. doi: 10.1182/blood-2010-02-258558

Supplementary files

Supplementary Files
Action
1. JATS XML
Download

Copyright (c) 2016 Savchenko A.A., Borisov A.G., Anisimova E.N., Belenyuk V.D., Kudryavtsev I.V., Reshetnikov I.V., Kvjatkovskaja S.V., Cejlikman V.J., Zorin A.N.

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.
СМИ зарегистрировано Федеральной службой по надзору в сфере связи, информационных технологий и массовых коммуникаций (Роскомнадзор).
Регистрационный номер и дата принятия решения о регистрации СМИ: серия ПИ № ФС 77 - 64788 от 02.02.2016.


This website uses cookies

You consent to our cookies if you continue to use our website.

About Cookies