A comparative analysis of in vitro expansion of natural killer cells of a patient with autoimmune haemolytic anaemia and ovarian cancer with patients with other solid tumours

Abstract

Abstract. The functional profile of natural killer (NK) cells has been reported to be lower in auto-immune haemolytic anaemia (AIHA). In this study, we report a comparative analysis of peripheral blood mononuclear cells (PBMNCs) and the in vitro expansion of NK cells in a patient with AIHA and cancer, with that of other cancer patients without AIHA. PBMNCs and in vitro NK-cell expansion of a 64-year old female patient with ovarian cancer and AIHA was compared with that of four other patients with cancer without AIHA who underwent autologous immune enhancement therapy (AIET). The NK cells were cultured using autologous plasma without feeder layers. The quantities of PBMNCs, NK cells and CD3-CD56+ cells were compared. The average quantity of PBMNCs per ml in Cases I to V were 10.71, 39.2, 49.26, 65.16 and 49.33x10 4 , respectively, and the average maximum count of NK cells was 3. 9, 1730.03, 1824.16, 1058.61 and 761x10 6 , respectively. The average percentage of CD3-CD56+ cells in Cases I to V following in vitro expansion was 1.2, 65.7, 28.63, 65.9 and 40%, respectively. In the present study, probably the first in the literature, the in vitro expansion of NK cells was found to be significantly lower in the AIHA patient. Previously, only a lower NK-cell functional profile was reported. Further studies are required to establish the association between AIHA and NK-cell profile and in vitro expansion, and to find common antibodies between red blood cells (RBCs) and NK cells.

Autologous Immune Enhancement Therapy in Philadelphia Chromosome Positive Acute Lymphoblastic Leukemia

“…LymphoPrep solution was used to isolate the PBMCs. The cell count was estimated using trypan blue dye-exclusion method and the NK cells were expanded for a period of 14 days using the methodology already reported [4][5][6][7]. Flow cytometry analysis was done to determine the percentage of NK cells employing markers like CD3, CD16 and CD56.…”

Pulmonary Tumor Thrombotic Microangiopathy

“…12 Furthermore, a high incidence of PTTM, up to 16.7%, has been described in the context of gastric carcinomas (3.3% in malignant tumors in general). 10 It is not well known why gastric carcinoma is more frequently associated with PTTM than other neoplasms. Interestingly, our 3 patients came from Latin American countries and were younger than most of the patients at gastric cancer diagnosis.…”

Autologous Immune Enhancement Therapy in Recurrent Ovarian Cancer with Metastases: A Case Report

“…The pellet containing white and red blood cells was mixed using phosphate buffered saline, and the routine procedure of density gradient centrifugation was done using LymphoPrep solution (GIBCO-BRL, Gaithersburg, Md., USA) to isolate the peripheral blood mononuclear cells (PBMNCs). The cell count was estimated using the Trypan blue dye-exclusion method and the cells were subjected to expansion as per the protocols described earlier [ 7 , 8 ] for a period of 10–16 days. Before cell transplantation, cells were tested for endotoxin levels using a Limulus Amebocyte Lysate kit (Charles River Laboratories, India), and flow cytometry analysis was done to determine the percentage of natural killer (NK) cells and activated T Lymphocytes employing markers viz., CD3, CD16 and CD56.…”