Development of apoptosis in Namalwa lymphoblastoid cells in conditions of clinorotation Nesterova N.V., Zagorodnya S.D., Golovan A.V., Baranova G.V. D.K. Zabolotny Institute of Microbiology and Virology NAS of Ukraine Laboratory of viruses reproduction

Infectious mononucleosis Clinical presentations of infectious mononucleosis (IM) are lymphadenitis, lymphadenopathy, adenoid disease, hepatitis (hepatosplenomegaly), splenomegaly, angina, hematological changes.

EBV-associated malignant lymphoproliferative diseases Burkitt's lymphoma Non- Hodgkin's lymphoma Nasopharyngeal carcinoma, Adenocarcinoma lymphogranulomatosis Oral cells lymphoplakia X-linked lymphoproliferative syndrome.

Autoimmune malignant Syndrome of a chronic fatigue. Autoimmune lymphoproliferative syndrome (ALPS) Pseudorheumatism

Tasks of research: 1. Study of influence of microgravitation on Namalwa lymphoid cells viability and functional condition 2. Study of effect microgravitation on Epstein-Barr virus reproduction in Namalwa cell culture. 3. Investigation of influence of microgravitation on apoptosis processes in Namalwa cell - Epstein-Barr virus system.

Suspension cell culture of B-cell line of the Burkitt's lymphoma – Namalwa - human malignant lymphoid cells were used as a test model in experiment with microgravitation. This cell line was given by bank of cell cultures of R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology of NAS of Ukraine. The cells were cultivated at the temperature of 37 0 С in media consisted of medium 1640 with supplement of 10 % heated embryonic serum, 2mM L-glutamine and antibiotics.

Namalwa cells were infected by Epstein-Barr virus (virus was purified by dextran sulphate density-gradient centrifugation, quantity of viral protein was determined using a «QuantiPro BCA ASSAY Kit», Sigma, USA). Quantity of DNA genome-equivalents was determined by PCR method in a system "AMPLY-Senc-100R" (Russia) and Programme " Biotest A ".

Horizontal clinorotation of cells was carried out at 37 0 С with speed 4 rpm (device «Clin - 1", "«Respirator", Donetsk) for modeling of microgravitation in ground conditions.

Influence of microgravitation on proliferate activity cells (MTT- test)

Level of Epstein-Barr virus core protein accumulation in Namalwa cell culture under the influence of microgravitation

Detection of apoptic manifestation in Namalwa cells using Hoechst (fluorescence microscopy) Apoptic fragmentation of nuclear cells

Influence of microgravitation on development of apoptosis in Epstein-Barr virus-infected Namalwa lymphoid cells (in percent). Time samplings 24 h48 h72 h Cells1073 Cells+EBV897 Cells+microgravitation Cells+EBV+microgravitation262736

Detection of apoptic manifestation in Namalwa cells using Annexin V and 6-Carboxyfluorescein (fluorescence microscopy) Annexin V - RED 6-Carboxyfluorescein – Green

Detection of apoptic manifestation in Namalwa cells using Acridine Orange and Ethidium Bromide (fluorescence microscopy)