İmatinib Tedavisi Alan Hastalarda T Lenfosit Aktivasyonunun Akım Sitometrik Analizi

Year 2020, Volume: 42 Issue: 2, 180 - 187, 17.03.2020

Deniz Goren Sahin Klara Dalva Sema Meriç Günhan Gürman Muhit Özcan Önder Arslan Mutlu Arat

https://doi.org/10.20515/otd.532364

Abstract

İmatinibin in vivo ortamda T lenfosit
fonksiyonları üzerine olan etkileri akım sitometrik olarak
değerlendirilmiştir.  Çalışmaya toplam 29 KML hastası ve 9 sağlıklı birey
dahil edilmiştir. KML hastaları kendi aralarında sırasıyla, yeni tanı almış ve
tedavi görmeyen, 1 yıldır imatinib alan ve 1 yıldan uzun süredir imatinib alan
hastalar olarak 3 gruba ayrılmıştır. CD4+ hücreler ve bu hücrelerdeki IL-4
ve IFNg ifadeleri, CD3+ T lenfositlerin % kaçının aktive edilmiş olduğu
(CD3+CD69+), CD8+ hücreler ve bu hücrelerde HLA-ABC ve HLA-DR ifade eden
hücrelerin oranı ve HLA-ABC ve HLA-DR ifadelerinin şiddeti değerlendirilmiştir. Gruplar arasında ortalama CD4 + hücre
sayısı açısından anlamlı fark yoktu. Bununla birlikte, kontrol grubunda daha
yüksek CD4 + hücrelerine doğru bir eğilim vardı. IL-4 ve IFN gama, gruplar
arasında istatistiksel olarak anlamlı bulunmadı. Kontrol grubu daha düşük IL-4
ve IFN gama ifade değerlerine sahipti. IL-4 ve IFN gama ifade etmeyen ortalama
CD4 + hücre sayısı kontrol grubunda diğer gruplara göre istatistiksel olarak
daha yüksekti. Kontrol grubunda ise % aktivasyon diğer gruplara göre azdı. Tüm
hasta gruplarında CD8 + hücre oranı istatistiksel olarak daha düşük bulundu (p
= 0,001). HLA-ABC ve HLA-DR'nin CD8 + hücrelerinde ekspresyonu gruplar arasında
benzerdi. İmatinibin sitokin sentezi üzerine herhangi bir etkisi
gözlenmemiştir. İmatinib tedavisi altındaki hastalarda T lenfosit
fonksiyonlarının etkilendiğine dair klinik belirgin gözlemler olmamakla
birlikte, subklinik etki varlığı araştırma konusudur. Yapılan in vitro çalışmaların in-vivo
korele olup olmadığının aydınlatılabilmesi için daha ileri çalışmalara
gereksinim vardır. 

Keywords

kronik miyelositer lösemi, akım sitometri, imatinib, T hücre

Flow Cytometric Analysis of T Lymphocyte Activation in CML Patients Under Imatinib Therapy

Abstract

To analyze
T cell functions by flow cytometry and to evaluate the possible functional
changes that might occur under imatinib therapy in CML patients. A total of 29
patients and 9 healthy control subjects were enrolled. Newly diagnosed patients
having no treatment (group 1), patients receiving imatinib for 1 year (group 2)
and patients receiving imatinib more than 1 year (group 3), healthy control
subjects (group 4). IL-4 and IFN gamma expression on CD4+ cells; how much
percentage of CD3+ T cells were activated (CD3+CD69+); CD8+ T cells and the
ratio and grade of expression of HLA-ABC and HLA-DR on those cells were
evaluated, respectively.

There was
no significant difference in terms of mean number of CD4+ cells between the
groups. However, there was a tendency towards higher CD4+ cells in control
group. IL-4 and IFN gamma were found not to be statistically significant
between the groups. Control group has lower IL-4 and IFN gamma expression
values. Mean number of CD4+ cells, which did not express IL-4 and IFN gamma,
were statistically higher in control group when compared to other groups. In
control group, % activation was decreased when compared to that of other
groups. CD8+ cell ratio was found to be statistically lower in all patient
groups (p=0.001). The expression of HLA-ABC and HLA-DR on CD8+ cells were
similar between the groups. We could not show any inhibitory effect of imatinib
on T cell functions in concordance with clinical experience and safety profile. 

Keywords

chronic myelogenous leukemia, flow cytometry, imatinib, T cell

References

• Shtivelman E, Lifshitz B, Gale RP, Canaani E. Fused transcript of abl and bcr genes in chronic myelogenous leukaemia. Nature. 1985;315(6020):550-4.
• Kurzrock R, Gutterman JU, Talpaz M. The molecular genetics of Philadelphia chromosome-positive leukemias. The New England journal of medicine. 1988;319(15):990-8.
• Lugo TG, Pendergast AM, Muller AJ, Witte ON. Tyrosine kinase activity and transformation potency of bcr-abl oncogene products. Science. 1990;247(4946):1079-82.
• Druker BJ, Tamura S, Buchdunger E, Ohno S, Segal GM, Fanning S, et al. Effects of a selective inhibitor of the Abl tyrosine kinase on the growth of Bcr-Abl positive cells. Nat Med. 1996;2(5):561-6.
• Kantarjian HM, O'Brien S, Cortes J, Giles FJ, Rios MB, Shan J, et al. Imatinib mesylate therapy improves survival in patients with newly diagnosed Philadelphia chromosome-positive chronic myelogenous leukemia in the chronic phase: comparison with historic data. Cancer. 2003;98(12):2636-42.
• Druker BJ, Guilhot F, O'Brien SG, Gathmann I, Kantarjian H, Gattermann N, et al. Five-year follow-up of patients receiving imatinib for chronic myeloid leukemia. N Engl J Med. 2006;355(23):2408-17.
• Barrett AJ. Mechanisms of the graft-versus-leukemia reaction. Stem Cells. 1997;15(4):248-58.
• Cwynarski K, Laylor R, Macchiarulo E, Goldman J, Lombardi G, Melo JV, et al. Imatinib inhibits the activation and proliferation of normal T lymphocytes in vitro. Leukemia. 2004;18(8):1332-9.
• Seggewiss R, Lore K, Greiner E, Magnusson MK, Price DA, Douek DC, et al. Imatinib inhibits T-cell receptor-mediated T-cell proliferation and activation in a dose-dependent manner. Blood. 2005;105(6):2473-9.
• Aswald JM, Lipton JH, Aswald S, Messner HA. Increased IFN-gamma synthesis by T cells from patients on imatinib therapy for chronic myeloid leukemia. Cytokines, cellular & molecular therapy. 2002;7(4):143-9.
• Pendergast AM, Muller AJ, Havlik MH, Maru Y, Witte ON. BCR sequences essential for transformation by the BCR-ABL oncogene bind to the ABL SH2 regulatory domain in a non-phosphotyrosine-dependent manner. Cell. 1991;66(1):161-71.
• Lim SH, Coleman S. Chronic myeloid leukemia as an immunological target. Am J Hematol. 1997;54(1):61-7.
• Clark RE, Christmas SE. BCR-ABL fusion peptides and cytotoxic T cells in chronic myeloid leukaemia. Leuk Lymphoma. 2001;42(5):871-80.
• Garicochea B, Chase A, Lazaridou A, Goldman JM. T lymphocytes in chronic myelogenous leukaemia (CML): no evidence of the BCR/ABL fusion gene detected by fluorescence in situ hybridization in 14 patients. Leukemia. 1994;8(7):1197-201.
• Gao H, Lee BN, Talpaz M, Donato NJ, Cortes JE, Kantarjian HM, et al. Imatinib mesylate suppresses cytokine synthesis by activated CD4 T cells of patients with chronic myelogenous leukemia. Leukemia. 2005;19(11):1905-11.
• Tsuda H, Yamasaki H. Type I and type II T cell profiles in chronic myelogenous leukemia. Acta haematologica. 2000;103(2):96-101.
• Leder C, Ortler S, Seggewiss R, Einsele H, Wiendl H. Modulation of T-effector function by imatinib at the level of cytokine secretion. Exp Hematol. 2007;35(8):1266-71.
• Dietz AB, Souan L, Knutson GJ, Bulur PA, Litzow MR, Vuk-Pavlovic S. Imatinib mesylate inhibits T-cell proliferation in vitro and delayed-type hypersensitivity in vivo. Blood. 2004;104(4):1094-9.
• Appel S, Balabanov S, Brummendorf TH, Brossart P. Effects of imatinib on normal hematopoiesis and immune activation. Stem cells (Dayton, Ohio). 2005;23(8):1082-8.
• Chen J, Schmitt A, Chen B, Rojewski M, Ringhoffer M, von Harsdorf S, et al. Imatinib impairs CD8+ T lymphocytes specifically directed against the leukemia-associated antigen RHAMM/CD168 in vitro. Cancer Immunol Immunother. 2007;56(6):849-61.
• Brauer KM, Werth D, von Schwarzenberg K, Bringmann A, Kanz L, Grunebach F, et al. BCR-ABL activity is critical for the immunogenicity of chronic myelogenous leukemia cells. Cancer research. 2007;67(11):5489-97.
• Mumprecht S, Matter M, Pavelic V, Ochsenbein AF. Imatinib mesylate selectively impairs expansion of memory cytotoxic T cells without affecting the control of primary viral infections. Blood. 2006;108(10):3406-13.